interleukin 8

C Subfamily
CC Subfamily
CXC Subfamily
CX3C Subfamily

Chemokines

gp130 (IL6ST) shared
IL13RA1 shared
IL3RB (CSF2RB) shared
ILRG shared

interleukin 18
interleukin 18 proprotein
interleukin 1 alpha
interleukin 1, alpha proprotein
interleukin 1 beta
interleukin 1, beta proprotein

interleukin 17
interleukin 17b
interleukin 17E
interleukin 17E isoform 1

IL-1 Family /IL-17 Family

interleukin 10
interleukin 19
interleukin 19 isoform 2
interleukin 20
interleukin 22
interleukin 24
interleukin 24 isoform 1
interleukin 28A
interleukin 28B
interleukin 29

IL-10 Family

interferon alpha family, gene 1
interferon, alpha 1
interferon beta
interferon, beta 1
interferon epsilon 1
interferon, epsilon 1
interferon gamma
interferon, gamma
interferon kappa
interferon, kappa
interferon, omega 1

Interferon Family

CSF1 Egf Flt3l
FLT3LG HGF Kitl
KITLG Pdgfa PDGFB
PDGFC Pdgfd PLEKHQ1
VEGF Vegfa VEGFB
VEGFC

PDGF Family

AMH Bmp2 Bmp7
GDF5 Inhba INHBB
INHBC INHBE Tgfb1
TGFB2 TGFB3

TGF-β Family

CD40LG Eda Fasl
FASLG Lta LTB
TNF Tnfsf10 Tnfsf11
TNFSF12 Tnfsf13 TNFSF13B
Tnfsf14 TNFSF18 TNFSF4
TNFSF7 TNFSF8 TNFSF9

TNF Family

INTERLEUKIN 8

LATEST LITERATURE

1: The European respiratory journal : official journal of the European Society for Clinical Respiratory Physiology, 2010 Sep 3, 411(17-18)
Signaling pathway of isophorone diisocyanate-responsive IL-8 in airway smooth muscle cells.

[Abstract]This study is first to analyse the soluble factors, secreted by the bronchial epithelium after exposure to isophorone diisocyanate (IPDI), responsible for increasing migration and proliferation of primary normal human bronchial smooth muscle cells (BSMC). We treated immortalized non-tumourigenic human bronchial epithelial cells (BEAS-2B) and primary normal human bronchial epithelial cells (HBEC) with IPDI, and then collected the condition medium (IPDI-BEAS-2B-CM and IPDI-HBEC-CM), which was added to BSMC. Exposure of BEAS-2B and HBEC to IPDI increased IL-8 production. Culture of BSMC with IPDI-BEAS-2B-CM and IPDI-HBEC-CM increased BSMC proliferation and migration, which are major features in asthma remodeling. Induction of BSMC proliferation and migration by IPDI-BEAS-2B-CM and IPDI-HBEC-CM was associated with increased FAK, Src, ERK1/2 and AKT activation. Blocking FAK by a specific inhibitor significantly decreased BSMC migration and proliferation by inhibiting ERK1/2 activation. FAK and ERK1/2 inhibitor also decreased IPDI-BEAS-2B-CM, IPDI-HBEC-CM and rhIL-8-mediated BSMC proliferation and migration, whereas blocking Rnd3 by siRNA failed to affect BSMC proliferation, suggesting that Rnd3 was only involved in the regulation of BSMC migration. Our study suggests that inhibition of IL-8 or IL-8-mediated FAK/ERK/Rnd3 signaling is an attractive therapeutic target for IPDI-mediated asthma.

2: Immunological investigations, 2010 Sep 1, 215(9-10)
Evaluation of Interleukin-8 in Hepatitis C Virus Infection: Relation to Combined peg-interferon Ribavirin Response and Genotype 4.

[Abstract]Hepatitis C virus (HCV), a major cause of liver disease worldwide, is frequently resistant to the antiviral alpha interferon (IFN). Previous studies have shown that HCV NS5A protein induces expression of the proinflammatory interleukin-8 (IL-8) and this may affect the therapeutic outcome. The aim of the present study was to investigate the relationship among levels of IL-8 in serum of subjects with past exposure to HCV indicated by positive IgG to HCV and negative PCR, patients with chronic HCV infections and in responder to combined alfa IFN and ribavirin therapy. The study included 48 Egyptian subjects with evidence of HCV infection. They were classified according to viremia to patients with chronic hepatitis C with positive viremia and immune subjects with positive HCV IgG alone. Chronic HCV patients were followed after therapy and 16 healthy adults as control group. It was found that viral load was dependent factor for the level of IL 8 (P = 0.003) and there was significant correlation between levels of IL-8 before treatment and after treatment. The present study highlights the kinetic of serum levels of interleukin-8 in patients with chronic hepatitis C genotype 4 before and after therapy with combined alfa interferon and ribavirin. It was demonstrated that HCV infection was associated with higher levels of interleukin-8 in pretreatment and posttreatment period. Moreover, immune subjects also had higher level of interleukin-8, indicating its role in immunity to HCV infection.

3: Journal of basic microbiology, 2010 Aug 30, 152(1-2)
Mutation of iceA in Helicobacter pylori compromised IL-8 induction from human gastric epithelial cells.

[Abstract]IceA of Helicobacter pylori (H. pylori) has been suggested as a virulence factor for the bacteria, but its pathogenic role remains unelucidated. Here, we examined the effect of iceA mutation on the secretion of IL-8 by human gastric epithelial cells. We also investigated whether the changes in IL-8 production caused by iceA mutation were associated with impaired adherence of H. pylori to the epithelial cells or with impaired apoptosis of these cells. The iceA mutant strain was constructed from wildtype H. pylori strain by insertional mutagenesis of iceA. The human gastric epithelial cells SGC7901 were infected with wildtype or mutant H. pylori for appropriate lengths of time. The adherence of the bacteria to the epithelial cells was examined by fluorescent microscopy using an anti-H. pylori antibody and flow cytometry. The apoptosis of the epithelial cells was studied by annexin-V staining and flow cytometry. The production of IL-8 by SGC 7901 cells was determined by ELISA. We found that iceA mutation was associated with significantly impaired production of IL-8 from the epithelial cells, which was not due to impaired adherence by the bacteria to the epithelial cells as wildtype and mutant H. pylori exhibited similar levels of binding to the epithelial cells. Furthermore, inactivation of iceA did not affect the apoptotic cell death of SGC7901. Our findings indicate that iceA may contribute to the pathogenicity of H. pylori by modulating the production of IL-8 by host epithelial cells. ((c) 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).

4: Environmental toxicology, 2010 Aug 27, 152(1-2)
Cement-related particles interact with proinflammatory IL-8 chemokine from human primary oropharyngeal mucosa cells and human epithelial lung cancer cell line A549.

[Abstract]Epidemiological studies have shown that respirable exposure to emitted cement particulate matter is associated with adverse health risk for human. The underlying mechanisms, however, are poorly understood. To examine the effect of cement, nine blinded cement-related particulates (<10 mum) were assessed with regard to their induction of the proinflammatory cytokines IL-6 and IL-8 in human primary epithelial cells (pEC) from oropharyngeal mucosa as well as from nonsmall-cell lung carcinoma (non-SCLC) cells A549. It was demonstrated that the cement specimens did not act cytotoxic as assessed by the lactate dehydrogenase (LDH) assay. The basal and IL-1beta-induced IL-8 expression was suppressed, in contrast to an unchanged IL-6. At the transcript level the basal and induced IL-6 and IL-8 gene expression was not influenced by cement dust. To discover the mechanism by which cement influenced the IL-8 expression the following experiments were performed. Submerse exposure experiments have shown that the release of IL-8 was suppressed by cement dust. Furthermore, the incubation of IL-8 with cement-related specimens under cell-free condition led to a loss of immunoreactive IL-8. An immunological masking of IL-8 by free soluble components of respiratory epithelial cells was excluded. Thus, the decrease of IL-8 protein content after cement exposure seems to be a result of the adsorption of IL-8 protein to cement particles and the inhibition of IL-8 release. In conclusion, due to absent cytotoxic and inflammatory effects of cement-related specimens in both human pEC and A549 cell models it remains open how cement exposure may lead to the respiratory adverse effects in humans. (c) 2010 Wiley Periodicals, Inc. Environ Toxicol, 2010.

5: Supportive care in cancer : official journal of the Multinational Association of Supportive Care in Cancer, 2010 Aug 28, 152(1-2)
The diagnostic value of CRP, IL-8, PCT, and sTREM-1 in the detection of bacterial infections in pediatric oncology patients with febrile neutropenia.

[Abstract]PURPOSE: In this study, we evaluated C-reactive protein (CRP), interleukin (IL)-8, procalcitonin (PCT), and soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) as predictors for bacterial infection in febrile neutropenia, plus their usefulness in febrile neutropenia during chemotherapy-induced gastrointestinal mucositis. METHODS: Plasma was obtained from pediatric oncology patients at presentation with febrile neutropenia (n = 43) and 24-48 h later (n = 17). The patients were classified as having or not having a bacterial infection. Plasma was also obtained of patients in the absence and in the presence of mucositis (n = 26). RESULTS: At presentation with febrile neutropenia, median IL-8 and PCT levels were significantly increased in patients with a bacterial infection, in contrast to CRP and sTREM-1. IL-8 was the most sensitive marker for the early detection of bacterial infection, in combination with clinical parameters or PCT the sensitivity reached 100%. After 24-48 h, only PCT was significantly elevated during bacterial infection. IL-8 levels were significantly increased during mucositis. Mucositis did not cause considerable changes in PCT levels. CONCLUSIONS: IL-8 is the most useful marker for the early detection of bacterial infections, compared with CRP, PCT, and sTREM-1. IL-8 in combination with clinical parameters or PCT might be even more useful. Gastrointestinal mucositis alone does not affect PCT levels, in contrast to IL-8 levels, and therefore, PCT might be more useful for the detection of bacterial infections during mucositis than IL-8.

6: Annals of allergy, asthma & immunology : official publication of the American College of Allergy, Asthma, & Immunology, 2010 Sep, 105(3)
Induction of interleukin 8 by American cockroach allergens from human airway epithelial cells via extracellular signal regulatory kinase and jun N-terminal kinase but not p38 mitogen-activated protein kinase.

[Abstract]BACKGROUND: Cockroaches are potent aeroallergens associated with asthma. Several reports suggest that a novel group of G protein-linked receptors, protease-activated receptors (PARs), may be involved in the intracellular signaling pathway induced by aeroallergens of the epithelial cells. OBJECTIVE: To investigate the mechanisms of American cockroach allergens (CraA) on interleukin 8 (IL-8) in human pulmonary epithelial cells. METHODS: Protease activities of CraA were quantified by the Azocoll method. The gene and protein expressions of IL-8 from CraA-stimulated A549 cells were quantified by real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The activity of different mitogen-activated protein kinases (MAPKs) was assessed by Western blot. RESULTS: CraA-induced A549 cell IL-8 secretion in a dose-dependent manner at both the messenger RNA and protein levels. CraA-induced IL-8 secretion can be blocked by serine protease inhibitors, phenylmethane sulfonyl fluoride, and aprotinin but not by other protease inhibitors. Blocking antibodies against the cleavage sites of PAR-2 and PAR-3, but not of PAR-1, inhibited CraA-induced IL-8 production. CraA induced significant PAR-2 and PAR-3 messenger RNA upregulation and extracellular-regulated kinase (ERK/1/2) and Jun N-terminal kinase (JNK) phosphorylation but not p38 MAPK. Furthermore, ERK1/2 (U0126) and JNK (SP600125) inhibitors inhibited CraA-induced IL-8 secretion by 100% and 45%, respectively. CONCLUSIONS: Both PAR-2 and PAR-3 might play a role in CraA-induced IL-8 secretion from human airway epithelial cells. It signals mainly through the ERK1/2 and partly from the JNK pathways. The key receptors and signaling molecules mediate cytokine release from the respiratory epithelium and can be potential therapeutic targets in treating cockroach allergy.

7: ORL; journal for oto-rhino-laryngology and its related specialties, 2010 Aug 21, 72(5)
Induction of IL-6 and IL-8 by House Dust Mite Allergen Der p1 in Cultured Human Nasal Epithelial Cells Is Associated with PAR/PI3K/NFkB Signaling.

[Abstract]Objective: The mechanism of action involved in how Dermatophagoides pteronyssinus (Der p) 1 initiates the nasal allergic cascade is poorly understood. Methods: We detected proinflammatory cytokine production (GM-CSF, TNF-alpha, IL-1beta, IL-6, and IL-8) and associated signal molecules in primarily cultured nasal epithelial cells (NECs) from patients with allergic rhinitis (AR) after Der p1 stimulation, using ELISA, RT-PCR, and Western blot. We also evaluated the importance of protease-activated receptors (PAR)/phosphatidylinositol 3 kinase (PI3K)/NFkappaB signaling pathways in IL-6 and IL-8 production using glucocorticoids and specific inhibitors, LY294002 and PDTC. Results: We observed significantly elevated IL-6 and IL-8 production (both gene and protein) in NECs after Der p1 stimulation, and demonstrated that the expressions of PAR2, pAkt, and pp65 were upregulated afterDer p1 stimulation, which were associated with IL-6 and IL-8 production in NECs. Conclusion: These findings demonstrate that the PAR/PI3K/NFkappaB signaling pathway is involved in the induction of IL-6 and IL-8 in Der-p1-stimulated NECs from AR patients, and may have potential implications for the prevention and treatment of AR and asthma.

8: Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society, 2010 Aug 19, 10(4)
Biomaterials modulate interleukin-8 and other inflammatory proteins during reepithelialization in cutaneous partial-thickness wounds in pigs.

[Abstract]ABSTRACT Acute and chronic cutaneous wounds remain a clinical challenge that require a mechanistic understanding to advance treatment options. For example, the role of inflammatory mediators during wound healing is not completely understood. Biomimetic materials, such as an in situ photopolymerizable semi-interpenetrating network (sIPN) derived from extracellular matrix components, show great potential in improving healing through the delivery of therapeutic agents and the function as a temporary tissue scaffold. In this study, we characterized the temporal profile of porcine cutaneous partial-thickness wound healing in response to Xeroform() and sIPN treatment via histological and inflammatory protein analyses in epidermal, remodeling dermal, and dermal regions. Generally, interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-10, IL-12p70, interferon-gamma, and tumor necrosis factor-alpha, but not IL-8, were expressed in the epidermis and remodeling dermis in a time course that followed the progression of epidermal maturation in response to both treatments. Differences in cellularity and protein expression were observed between treatments in a time- and region-dependent manner. In particular, the healing response to sIPN exemplified a potentially key relationship between IL-8 expression and reepithelialization. These results provide insights into the expression of inflammatory mediators and the time course of cutaneous healing and the capacity for biomaterials to further modulate this relationship.

9: Journal of agricultural and food chemistry, 2010 Aug 19, 13(8)
Oenothera paradoxa Defatted Seeds Extract and Its Bioactive Component Penta-O-galloyl-beta-d-glucose Decreased Production of Reactive Oxygen Species and Inhibited Release of Leukotriene B(4), Interleukin-8, Elastase, and Myeloperoxidase in Human Neutrophi

[Abstract]In this study, we analyzed ex vivo the effect of an aqueous extract of Oenothera paradoxa defatted seeds on the formation of neutrophil-derived oxidants. For defining active compounds, we also tested lypophilic extract constituents such as gallic acid, (+)-catechin, ellagic acid, and penta-O-galloyl-beta-d-glucose and a hydrophilic fraction containing polymeric procyanidins. The anti-inflammatory potential of the extract and compounds was tested by determining the release from activated neutrophils of elastase, myeloperoxidase, interleukin-8 (IL-8), and leukotriene B(4) (LTB(4)), which are considered relevant for the pathogenesis of cardiovascular diseases. The extract of O. paradoxa defatted seeds displays potent antioxidant effects against both 4beta-phorbol-12beta-myristate-alpha13-acetate- and formyl-met-leu-phenylalanine-induced reactive oxygen species production in neutrophils with IC(50) values around 0.2 mug/mL. All types of polyphenolics present in the extract contributed to the extract antioxidant activity. According to their IC(50) values, penta-O-galloyl-beta-d-glucose was the more potent constituent of the extract. In cell-free assays, we demonstrated that this effect is partially due to the scavenging of O(2)(-) and H(2)O(2) oxygen species. The extract and especially penta-O-galloyl-beta-d-glucose significantly inhibit elastase, myeloperoxidase IL-8, and LTB(4) release with an IC(50) for penta-O-galloyl-beta-d-glucose of 17 +/- 1, 15 +/- 1, 6.5 +/- 2.5, and around 20 muM, respectively. The inhibition of penta-O-galloyl-beta-d-glucose on reactive oxygen species and especially on O(2)(-) production, myeloperoxidase, and chemoattractant release may reduce the interaction of polymorphonuclear leukocyte with the vascular endothelium and by that potentially diminish the risk of progression of atherosclerosis development.

10: Biology of reproduction, 2010 Aug 18, 10(4)
The Chemokine IL8 Is Up-Regulated in Bovine Endometrial Stromal Cells by the BoHV-4 IE2 Gene Product, ORF50/Rta: A Step Ahead Toward a Mechanism for BoHV-4 Induced Endometritis.

[Abstract]Postpartum infections of the endometrium and metritis are common causes of delayed conception and infertility in cattle. These infections are characterized by inflammation of the endometrium and secretion of the chemokine interleukin 8 (IL8), which attracts granulocytes to the endometrium. Bovine herpesvirus 4 (BoHV-4) is tropic for the endometrium, and the only virus consistently associated with postpartum metritis. The BoHV-4 IE2 gene is the first viral gene transcribed by host cells after infection and the IE2 gene product, ORF50/Rta, transactivates host cell genes. The present study tested the hypothesis that ORF50/Rta transactivates the IL8 gene promoter during BoHV-4 infection of bovine endometrial stromal cells (BESCs). Infection of primary BESCs with BoHV-4 stimulated IL8 gene promoter activity and IL8 protein secretion. However, IL8 production was dependent on the transcription of viral genes as psoralen/UV crosslinking of the viral DNA abrogated the response to BoHV-4 infection. Furthermore, IL8 promoter serial deletion analysis revealed a specific region responsive to ORF50/Rta. These observations may represent an endometrial defense mechanism against viral infection or a virulence mechanism by which viral replication stimulates chemokine secretion to attract more susceptible host cells to the endometrium.

11: Protein science : a publication of the Protein Society, 2010 Aug 17, 10(4)
Generation of high-affinity fully human anti-interleukin-8 antibodies from its cDNA by two-hybrid screening and affinity maturation in yeast.

[Abstract]We have developed a technology for rapidly generating novel and fully human antibodies by simply using the antigen DNA. A human single-chain (scFv) antibody library was constructed in a yeast two-hybrid vector with high complexity. After cloning cDNA encoding the mature sequence of human interleukin-8 (hIL8) into the yeast two-hybrid system vector, we have screened the human scFv antibody library, and obtained 3 distinct scFv clones that could specifically bind to hIL8. One clone was chosen for further improvement by a novel affinity maturation process using the error-prone PCR of the scFv sequence followed by additional rounds of yeast two-hybrid screening. The scFv antibodies of both primary and affinity matured scFv clones were expressed in E. coli. All purified scFvs showed specific binding to hIL8 in reciprocal co-immunoprecipitation and ELISA assays. All scFvs, as well as a fully human IgG antibody converted from one of the scFv clones and expressed in the mammalian cells, were able to effectively inhibit hIL8 in neutrophil chemotaxis assays. The technology described can generate fully human antibodies with high efficiency and low cost.

12: Experimental lung research, 2010 Sep, 36(7)
Differential effects of human neutrophil peptide-1 on growth factor and interleukin-8 production by human lung fibroblasts and epithelial cells.

[Abstract]ABSTRACT alpha-Defensins, antimicrobial peptides produced mainly by neutrophils, have been reported to be associated with a wide variety of lung diseases, including idiopathic pulmonary fibrosis (IPF), cystic fibrosis (CF), and diffuse panbronchiolitis (DPB). In each disease, alpha-defensins are located in different areas, such as around the alveolar septa in IPF and around the airways in CF and DPB, suggesting that alpha-defensins play different roles. Meanwhile, growth factors are known to contribute to IPF, CF, and DPB. alpha-Defensins are known to induce interleukin (IL)-8 in airway epithelial cells, but the effects of alpha-defensins on the release of growth factors from various components in the lung have not been sufficiently investigated. In the present study, the in vitro effects of human neutrophil peptide (HNP)-1 (a subtype of alpha-defensin) on the expressions of IL-8 and growth factors in lung fibroblasts, bronchial epithelial cells, and alveolar epithelial cells were examined. HNP-1 mainly enhanced the expression of IL-8 in epithelial cells, whereas it enhanced transforming growth factor-beta and vascular endothelial growth factor expressions in lung fibroblasts. These results suggest that alpha-defensins play different roles in the pathogenesis of IPF, CF, and DPB according to the location in the lung where the alpha-defensins are mainly produced.

13: PloS one, 2010, 5(8)
Androgen-regulated expression of arginase 1, arginase 2 and interleukin-8 in human prostate cancer.

[Abstract]BACKGROUND: Prostate cancer (PCa) is the most frequently diagnosed cancer in North American men. Androgen-deprivation therapy (ADT) accentuates the infiltration of immune cells within the prostate. However, the immunosuppressive pathways regulated by androgens in PCa are not well characterized. Arginase 2 (ARG2) expression by PCa cells leads to a reduced activation of tumor-specific T cells. Our hypothesis was that androgens could regulate the expression of ARG2 by PCa cells. METHODOLOGY/PRINCIPAL FINDINGS: In this report, we demonstrate that both ARG1 and ARG2 are expressed by hormone-sensitive (HS) and hormone-refractory (HR) PCa cell lines, with the LNCaP cells having the highest arginase activity. In prostate tissue samples, ARG2 was more expressed in normal and non-malignant prostatic tissues compared to tumor tissues. Following androgen stimulation of LNCaP cells with 10 nM R1881, both ARG1 and ARG2 were overexpressed. The regulation of arginase expression following androgen stimulation was dependent on the androgen receptor (AR), as a siRNA treatment targeting the AR inhibited both ARG1 and ARG2 overexpression. This observation was correlated in vivo in patients by immunohistochemistry. Patients treated by ADT prior to surgery had lower ARG2 expression in both non-malignant and malignant tissues. Furthermore, ARG1 and ARG2 were enzymatically active and their decreased expression by siRNA resulted in reduced overall arginase activity and l-arginine metabolism. The decreased ARG1 and ARG2 expression also translated with diminished LNCaP cells cell growth and increased PBMC activation following exposure to LNCaP cells conditioned media. Finally, we found that interleukin-8 (IL-8) was also upregulated following androgen stimulation and that it directly increased the expression of ARG1 and ARG2 in the absence of androgens. CONCLUSION/SIGNIFICANCE: Our data provides the first detailed in vitro and in vivo account of an androgen-regulated immunosuppressive pathway in human PCa through the expression of ARG1, ARG2 and IL-8.

14: Zhongguo fei ai za zhi = Chinese journal of lung cancer, 2010 Aug 20, 13(8)
[Overexpression of IL-8 and MMP-9 Confer High Malignant Phenotype in Patients with Non-small Cell Lung Cancer.]

[Abstract]BACKGROUND: IL-8 (interleukin-8) has been identified as a chemotactic factor, but recent found that IL-8 and matrix metalloproteinase-9 (MMP-9) are important cytokines which are closely related to the growth and metastasis of tumor. The aim of this study is to explore the relationship between IL-8, MMP-9 expressions and clinical pathological features of non-small cell lung cancer (NSCLC) patients and evaluate the diagnostic potential of IL-8, MMP-9 as tumor markers. METHODS: The serum levels of IL-8 and MMP-9 were detected by enzyme-linked immunosorbentassay (ELISA) in 141 NSCLC patients, 40 healthy adults and 40 patients with benign pulmonary disease. The expressions of IL-8 and MMP-9 were detected by immunohistochemical method in 95 NSCLC tissues, and 21 benign disease lung tissues, 25 normal lung tissues as control. RESULTS: The level of expression of IL-8 and MMP-9 in serum and tissue of NSCLC was significantly higher than that of healthy and benign respiratory disease, and the expression was gradually increased with the upgrade of clinicopathological stage. The serum and tissue expression of IL-8 and MMP-9 in NSCLC patients with lymph node metastasis was remarkably higher than that without lymph node metastasis. There is an positive correlation (r=0.765) between IL-8 and MMP-9 in the tissue of NSCLC patients. CONCLUSIONS: This study has confirmed that IL-8, MMP-9 expressions are related to the development of NSCLC. There is an obvious correlation between IL-8 expression and lymph node metastasis, IL-8 may facilitate the lymph node metastasis by up-regulating MMP-9 expression. Serum level of IL-8 is a valuable auxiliary parameter in diagnosing lymph node metastases of NSCLC with good sensitivity and specificity.

15: International immunopharmacology, 2010 Jul 30, 121(1-3)
Ginsenoside Rb1 and paeoniflorin inhibit transient receptor potential vanilloid-1-activated IL-8 and PGE(2) production in a human keratinocyte cell line HaCaT.

[Abstract]Ginsenoside Rb1 (GRb1) and paeoniflorin (PF) are active substances of Chinese traditional herbs and have been commonly used to treat skin inflammation diseases, but little is known about the mechanisms involved. Transient receptor potential vanilloid-1 (TRPV1) was originally identified on neuronal cells, and later it was also found on non-neuronal cells, such as keratinocytes. In the present study, the effects of GRb1 and PF on the production of inflammatory mediators and the possible mechanisms of TRPV1 in these mediators induction were explored. It has been shown that GRb1 and PF inhibited the productions of IL-8 and PGE(2) induced by capsaicin (CAP) in keratinocyte HaCaT cells and HEK 293T-TRPV1 cells (which were transgenic and overexpressed TRPV1) but had no effect on HEK 293T mock cells (p<0.05). Besides, CAP was able to induce calcium influx and nuclear factor kappa B(NF-kappaB) transcriptional activity in HaCaT cells and HEK 293T-TRPV1 cells, but had no effect on HEK 293T mock cells. Furthermore, GRb1 inhibited CAP-induced calcium influx and NF-kappaB transcriptional activity in both HaCaT cells and HEK 293T-TRPV1 cells. However, PF decreased CAP-induced calcium influx and NF-kappaB transcriptional activity only in HaCaT cells. This would suggest that GRb1 inhibits CAP-induced calcium influx and NF-kappaB activity through TRPV1 signal, while calcium influx and NF-kappaB activity might not be involved in the inhibitory effect of PF on TRPV1 signal. Furthermore, the inhibitory rates of GRb1 and PF on IL-8 and PGE(2) production were higher than those caused by capsazepine, an antagonist of TRPV1, suggesting that GRb1 and PF have great potential in clinical treatment of skin diseases.

16: Rheumatology international, 2010 Jul 28, 121(1-3)
Nicotine inhibits tumor necrosis factor-alpha induced IL-6 and IL-8 secretion in fibroblast-like synoviocytes from patients with rheumatoid arthritis.

[Abstract]It was recently demonstrated that the cholinergic anti-inflammatory pathway can modulate host inflammatory responses via cholinergic mediators or via electrical stimulation of the vagus nerve. Here, we investigated whether nicotine, a selective cholinergic agonist, plays any anti-inflammatory role in rheumatoid arthritis fibroblast-like synoviocytes (FLS). We observed that low concentrations (0.1-100 muM) of nicotine did not affect FLS viability in lactate dehydrogenase release test or the MTT assay. Nicotine at concentrations of 0.1-10 muM dose reduced the protein and mRNA expression of IL-6 and IL-8 induced by tumor necrosis factor-alpha (TNFalpha). Nicotine also inhibited nuclear factor (NF)-kappaB (p65) translocation from the cytoplasm to the nucleus, based on Western blotting and immunocytochemical analysis. In conclusion, nicotine can inhibit the TNFalpha dependant inflammatory pathway in synoviocytes by suppressing the activation of the NF-kappaB pathway.

17: International journal of cancer. Journal international du cancer, 2010 Jul 20, 121(1-3)
Interleukin-8 is associated with proliferation, migration, angiogenesis and chemosensitivity in vitro and in vivo in colon cancer cell line models.

[Abstract]Interleukin-8 (IL-8), a chemokine with a defining CXC amino acid motif, is known to possess tumorigenic and proangiogenic properties. Overexpression of IL-8 has been detected in many human tumors, including colorectal cancer, and is associated with poor prognosis. The goal of our study was to determine the role of IL-8 overexpression in colorectal cancer cells in vitro and in vivo. We stably transfected the IL-8 cDNA into two human colon cancer cell lines, HCT116 and Caco2, and selected IL-8-secreting transfectants. Real time RT-PCR confirmed that IL-8 mRNA was overexpressed in IL-8 transfectants with 45 approximately 85-fold higher than parental cells. The IL-8-transfected clones secreted 19 approximately 28-fold more IL-8 protein than control and parental cells as detected by ELISA. The IL-8 transfectants demonstrated increased cellular proliferation, cell migration and invasion based on functional assays. Growth inhibition studies showed that IL-8 overexpression lead to a significant resistance to oxaliplatin (P < 0.0001). Inhibition of IL-8 overexpression with small interfering RNA reversed the observed increases in tumorigenic functions and oxaliplatin resistance suggesting that IL-8 not only provides a proliferative advantage, but also promotes the metastatic potential of colon cancer cells. Using a tumor xenograft model, IL-8-expressing cells formed significantly larger tumors than the control cells with increased microvessel density. Together, these findings indicate that overexpression of IL-8 promotes tumor growth, metastasis, chemoresistance and angiogenesis, implying IL-8 to be an important therapeutic target in colorectal cancers.

18: Respiratory research, 2010 Jul 13, 11(1)
Pneumocystis cell wall beta-glucan stimulates calcium-dependent signaling of IL-8 secretion by human airway epithelial cells.

[Abstract]ABSTRACT: BACKGROUND: Respiratory failure secondary to alveolar inflammation during Pneumocystis pneumonia is a major cause of death in immunocompromised patients. Neutrophil infiltration in the lung of patients with Pneumocystis infection predicts severity of the infection and death. Several previous studies indicate that airway epithelial cells release the neutrophil chemoattractant proteins, MIP-2 (rodents) and IL-8 (humans), in response to Pneumocystis and purified Pneumocystis cell wall beta-glucans (PCBG) through the NF-kappaB-dependent pathway. However, little is known about the molecular mechanisms that are involved in the activation of airway epithelium cells by PCBG resulting in the secretion of IL-8. METHOD: To address this, we have studied the activation of different calcium-dependent mitogen-activated protein kinases (MAPKs) in 1HAEo- cells, a human airway epithelial cell line. RESULTS: Our data provide evidence that PCBG induces phosphorylation of the MAPKs, ERK, and p38, the activation of NF-kappaB and the subsequently secretion of IL-8 in a calcium-dependent manner. Further, we evaluated the role of glycosphingolipids as possible receptors for beta-glucans in human airway epithelial cells. Preincubation of the cells with D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) a potent inhibitor of the glycosphingolipids synthesis, prior to PCBG stimulation, significantly decreased IL-8 production. CONCLUSION: These data indicate that PCBG activates calcium dependent MAPK signaling resulting in the release of IL-8 in a process that requires glycosphingolipid for optimal signaling.

19: Cytokine, 2010 Aug, 51(2)
The interleukin-8-251 AA genotype is associated with angiogenesis in gastric carcinogenesis in Helicobacter pylori-infected Koreans.

[Abstract]Helicobacter pylori (H. pylori) is an important risk factor of gastric adenocarcinoma. Interleukin (IL)-8 is a potent angiogenic factor and plays an important role in inflammation of gastric mucosa by H. pylori. Host susceptibility may help to predict H. pylori-infected individuals with a higher risk of gastric adenocarcinoma. The aim of this study was to clarify the effect of IL-8 polymorphism on angiogenesis in the process of gastric carcinogenesis in H. pylori-infected Koreans. The IL-8-251A/T polymorphism was genotyped by PCR-RFLP from a total of 395 subjects; 92 normal controls, 87 H. pylori-infected controls, 108 chronic atrophic gastritis and/or intestinal metaplasia and 108 adenocarcinoma. The gastric mucosal concentrations of IL-8, membrane metalloproteinase (MMP)-9, angiopoietin (Ang)-1, and vascular endothelial growth factor (VEGF) were measured by ELISA. MMP-9 concentrations were increased with disease progression. There was significant correlation between MMP-9 and disease progression in AA (r=0.42, p<0.01) and AT genotype (r=0.43, p<0.01). Ang-1 concentrations were increased in AA genotype (r=0.40, p=0.01). However, there was no significant correlation between VEGF and disease progression in AA genotype. In conclusion, IL-8-251 AA genotype may be associated with angiogenesis in gastric carcinogenesis in H. pylori-infected Koreans.

20: Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society, 2010 Jul 9, 285(28)
Stability of interleukin 8 and neutrophil elastase in bronchoalveolar lavage fluid following long-term storage.

[Abstract]BACKGROUND: Interleukin-8 (IL-8) and neutrophil elastase (NE) are commonly measured markers of inflammation in bronchoalveolar lavage (BAL) fluid from patients with cystic fibrosis. Longitudinal analysis assumes uniform stability during storage, however the effect of extended low-temperature storage on these markers remains unclear. METHODS: BAL fluid from 104 children with cystic fibrosis was assayed for IL-8 and NE after storage at 4 degrees C for 7days and -80 degrees C for up to 6years and compared with the initial assays performed soon after collection. RESULTS: IL-8 levels were stable after any measured length of time at -80 degrees C or 4 degrees C. NE levels were stable for 6months at -80 degrees C but decreased beyond that or after 7days at 4 degrees C. CONCLUSIONS: Our data support the stability of IL-8 in BAL stored at -80 degrees C for prolonged periods. NE in BAL decreases with storage and should be assayed as soon as practical after collection.

21: Cellular microbiology, 2010 Jul 8, 285(28)
Positive transcription elongation factor b (P-TEFb) contributes to dengue virus-stimulated induction of interleukin-8 (IL-8).

[Abstract]Summary Dengue virus (DENV) is one of the most common infectious pathogens worldwide. One major clinical and pathogenic feature of DENV infection is the elevation of interleukin-8 (IL-8) expression; however, little is known about the molecular mechanism of DENV-induced chemokine production. The positive transcription elongation factor b (P-TEFb) composed of CDK9 and cyclin T1 stimulates gene expression by enhancing RNA polymerase II (RNA pol II) processivity. This study examined the possibility that P-TEFb mediates DENV-induced IL-8 expression. The treatment of either a pharmacological inhibitor of P-TEFb, 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole (DRB) or cyclin T1 siRNA prior to DENV infection abolished the elevation of IL-8, indicating that P-TEFb is essential for IL-8 induction. Moreover, DENV core protein participated in the activation of IL-8 promoter in a P-TEFb-dependent manner. Immunostaining and co-immunoprecipitation assays demonstrated the association between P-TEFb and DENV core protein. Finally, chromatin immunoprecipitation (ChIP) results indicated that P-TEFb and DENV core protein were recruited to the transcriptionally active IL-8 gene promoter. Taken together, this study showed that P-TEFb and DENV core protein work in concert to enhance IL-8 gene expression by DENV infection. This is the first demonstration of P-TEFb being directly involved in virus-induced host gene expression by interacting with a viral structural protein.

22: Immunobiology, 2010 September -, 215(9-10)
The novel RUNX3/p33 isoform is induced upon monocyte-derived dendritic cell maturation and downregulates IL-8 expression.

[Abstract]RUNX proteins are heterodimeric factors that play crucial roles during development and differentiation of cells of the immune system. The RUNX3 transcription factor controls lineage decisions during thymopoiesis and T-cell differentiation, and modulates myeloid cell effector functions. We now report the characterization of the human RUNX3/p33 isoform, generated by splicing out a Runt DNA-binding domain-encoding exon, and whose transcriptional activities differ from those of the prototypic RUNX3/p44 molecule. Unlike RUNX3/p44, RUNX3/p33 is induced upon maturation of monocyte-derived dendritic cells (MDDC), and is unable to transactivate the regulatory regions of the CD11a, CD11c and CD49e integrin genes. Overexpression of RUNX3/p33 in myeloid cell lines led to diminished expression of genes involved in inflammatory responses. Moreover, overexpression of RUNX3/p33 down-modulated the basal level of IL-8 production from immature monocyte-derived dendritic cells (MDDC). Besides, siRNA-mediated knock-down of RUNX3 led to diminished levels of IL-8 RNA in immature MDDC, and modulated the neutrophil-recruiting capacity of myeloid cell line supernatants. Since IL-8 promotes neutrophil chemotaxis and degranulation during inflammatory responses, and exerts mitogenic and angiogenic actions within tumor microenvironment, our results imply that myeloid RUNX3 expression regulates the recruitment of leukocytes towards inflammatory foci and might also contribute to human cancer progression.

23: Biochemical pharmacology, 2010 Jul 5, 285(28)
Decoy oligodeoxyribonucleotides and peptide nucleic acids-DNA chimeras targeting nuclear factor kappa-B: inhibition of IL-8 gene expression in Cystic Fibrosis cells infected with Pseudomonas aeruginosa.

[Abstract]Cystic fibrosis (CF) is characterized by a deep inflammatory process, with production and release of cytokines and chemokines, among which interleukin 8 (IL-8) represents one of the most important. Accordingly, there is a growing interest in developing therapies against IL-8, with the aim of reducing the excessive inflammatory response in the airways of CF patients. Since transcription factor NF-kappaB plays a critical role in IL-8 expression, the transcription factor decoy (TFD) strategy might be of interest. TFD is based on biomolecules mimicking the target sites of transcription factors (TFs) and able to interfere with TF activity when delivered to target cells. Here we review the inhibitory effects of decoy oligodeoxyribonucleotides (ODNs) on expression of IL-8 gene and secretion of IL-8 by cystic fibrosis cells infected by Pseudomonas aeruginosa. In addition, the effects of decoy molecules based on peptide nucleic acids (PNAs) are discussed. In this respect PNA-DNA-PNA (PDP) chimeras are interesting: (a) unlike PNAs, they can be complexed with liposomes and microspheres; (b) unlike oligodeoxyribonucleotides (ODNs), they are resistant to DNAses, serum and cytoplasmic extracts; (c) unlike PNA/PNA and PNA/DNA hybrids, they are potent decoy molecules. Interestingly, PDP/PDP NF-kappaB decoy chimeras inhibit accumulation of pro-inflammatory mRNAs (including IL-8 mRNA) in Pseudomonas aeruginosa infected IB3-1 cells reproducing the effects of decoy oligonucleotides. The effect of PDP/PDP chimeras, unlike ODN-based decoys, are observed even in absence of protection with lipofectamine. Since IL-8 is pivotal in pro-inflammatory processes affecting cystic fibrosis, inhibition of its functions might have a clinical relevance.

24: Placenta, 2010 Jun 30, 72(1)
Stretch and inflammation-induced Pre-B cell colony-enhancing factor (PBEF/Visfatin) and Interleukin-8 in amniotic epithelial cells.

[Abstract]Preterm birth continues to be a growing problem in the USA. Although approximately half of preterm births are caused by intrauterine infection, uterine over-distension is also a cause. In this study we have compared the effects of static stretch, cyclic stretch/release and an inflammatory stimulus alone and in combination on the expression of Pre-B cell colony-enhancing factor (PBEF) and IL-8 in primary amniotic epithelial cells (AEC). We then sought to identify some of the mechanism(s) by which these cells respond to stretching stimuli. We show that cyclic stretch/release is a more robust stimulus for both PBEF and IL-8 than static stretch. Cyclic stretch/release increased both intracellular and secreted PBEF and a combination of both types of stretch was a more robust stimulus to PBEF that IL-8. However, when an inflammatory stimulus (IL-1beta) was added to either kind of stretch, the effect on IL-8 was much greater than that on PBEF. Thus, different kinds of stretch affect the expression of these two cytokines from AEC, but inflammation is a much stronger stimulus of IL-8 than PBEF, agreeing with its primary role as a chemokine. Although the AEC showed morphological signs of increased cellular stress during stretching, blocking reactive oxygen species (ROS) had little effect. However, blocking integrin binding to fibronectin significantly reduced the responses of both PBEF and IL-8 to cyclic stretch/release. The increased PBEF, both intracellularly and secreted, suggests that it functions both to increase the metabolism of the cells, at the same time as stimulating further the cytokine cascade leading to parturition.

25: Arteriosclerosis, thrombosis, and vascular biology, 2010 Jul 1, 182(1)
Lead Contributes to Arterial Intimal Hyperplasia Through Nuclear Factor Erythroid 2-Related Factor-Mediated Endothelial Interleukin 8 Synthesis and Subsequent Invasion of Smooth Muscle Cells.

[Abstract]OBJECTIVE: To validate the hypothesis that the toxic heavy metal lead (Pb) may be linked to cardiovascular diseases both in vitro and in vivo in the initiation phase of atherosclerosis (ie, intimal hyperplasia). METHODS AND RESULTS: During the human study part of this project, serum Pb levels of healthy young females were correlated to carotid intima-media thickness. Multivariate logistic regression analyses showed that increased serum Pb levels were significantly associated with an increased intima-media thickness (P=0.01; odds ratio per SD unit, 1.6 [95% CI, 1.1 to 2.4]). In vitro, Pb induced an increase in interleukin 8 production and secretion by vascular endothelial cells. Nuclear factor erythroid 2-related factor-2 is the crucial transcription factor involved in Pb-induced upregulation of interleukin 8. Endothelial cell-secreted interleukin 8 triggered intimal invasion of smooth muscle cells and enhanced intimal thickening in an arterial organ culture model. This phenomenon was further enhanced by Pb-increased elastin synthesis of smooth muscle cells. CONCLUSIONS: Our data support the hypothesis that Pb is a novel, independent, and significant risk factor for intimal hyperplasia.

26: Investigative ophthalmology & visual science, 2010 Jun 30, 72(1)
Multiplex Cytokine Analysis Reveals Elevated Concentration of Interleukin-8 in Glaucomatous Aqueous Humor.

[Abstract]Purpose. To test the hypothesis that immune activation occurs in glaucoma by comparing concentrations of multiple cytokines in aqueous humor (AH) from patients with primary open angle glaucoma (POAG) and from cataract patients without glaucoma as controls. Methods. Cytokine concentrations in AH obtained during surgery were measured using microparticle-based immunoassays. Localized expression of IL-8 protein was investigated by immunohistochemistry of human eyes. Results. Eight cytokines [interleukin 1beta (IL-1beta), IL-2, IL-4, IL-5, IL-10, IL-12, interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha)] were below limits of detection and 2 cytokines (IL-18 and IL-15) were detected at low levels and/or in only a few patients. Although IL-6 was detected in 26 of 30 control patients (median 2.7 pg/ml) and 23 of 29 POAG patients (median 1.6 pg/ml), the difference was not statistically significant. IL-8 was detected in 28 of 30 control patients (median 1.8 pg/ml) and in all 29 POAG patients (median 4.9 pg/ml). The higher IL-8 concentration in the AH of POAG patients was statistically significant (p<0.001). In pairs of eyes from patients with asymmetrical glaucomatous optic nerve damage, IL-8 concentration was higher in AH of the more severely affected eye (p<0.05). Patients with severe visual field defects had higher IL-8 concentrations in the AH than did patients with mild visual field defects. IL-8 protein expression was found in human retina and optic nerve. Conclusions: Concentration of the inflammatory cytokine IL-8 is significantly elevated in AH of POAG patients, supporting the hypothesis that immune activation occurs in glaucoma.

27: Clinical and vaccine immunology : CVI, 2010 Jun 30, 72(1)
IL-8 Production by Human Airway Epithelial Cells in Response to Pseudomonas aeruginosa Clinical Isolates Expressing a-Type or b-Type Flagellins.

[Abstract]Pseudomonas aeruginosa lung infection is a major cause of morbidity and mortality worldwide. P. aeruginosa flagellin, the main structural protein of the flagellar filament, is a virulence factor with proinflammatory activity on respiratory epithelial cells. P. aeruginosa bacteria express one of two isoforms of flagellin (a-type or b-type) that differ in their primary amino acid sequences as well as in post-translational glycosylation. In this study, the distribution of a- and b-type flagellins among 3 P. aeruginosa laboratory strains and 14 clinical isolates (1 ulcerative keratitis, 3 cystic fibrosis, and 10 acute pneumonia) was determined, and their ability to stimulate IL-8 production by human airway epithelial cells was compared. By comparison with the PAK (a-type) and PAO1 (b-type) prototype laboratory strains, 10/14 (71.4%) of clinical isolates expressed a-type and 4/14 (28.6%) expressed b-type flagellins. Among 4 cell lines surveyed, BEAS-2B cells were found to give the greatest difference between constitutive and flagellin-stimulated IL-8 production. All 17 flagellins stimulated IL-8 production by BEAS-2B cells (range, 700-4,000 pg/ml). However, no discernable differences in IL-8 production were evident when comparing a-type vs. b-type flagellins, or flagellins from laboratory vs. clinical strains or amongst the clinical strains.

28: The Journal of biological chemistry, 2010 Jun 30, 72(1)
Peroxisome Proliferator-activated receptor (PPAR) {delta} activators induce IL-8 expression in non-stimulated endothelial cells in a transcriptonal and posttranscriptional manner.

[Abstract]Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors that are implicated in the regulation of lipid and glucose homeostasis. PPAR agonists have been shown to control inflammatory processes, in part by inhibiting distinct pro-inflammatory genes (e.g., Il-1beta and IFN-gamma). IL-8 is a member of the pro-inflammatory chemokine family that are important for various functions, such as mediating the adhesion of eosinophilic granulocytes onto endothelial cells. The influence of PPARdelta activators on the expression of IL-8 in non-induced quiescent endothelial cells is unclear. Therefore, we explored the influence of PPARdelta activators on the expression of IL-8 in non-stimulated endothelial cells. PPARdelta agonists induce IL-8 expression in HUVECs. This induction is demonstrated at the level of both protein and mRNA expression. Transcriptional activation studies using IL-8 reporter gene constructs, and DNA binding assays revealed that PPARdelta agonists mediated their effects via an NFkappaB binding site. It is well known that IL-8 is also regulated by mRNA stability. To provide further evidence for this concept, we performed mRNA stability assays and found that PPARdelta agonists induce the mRNA stability of IL-8. In addition, we showed that PPARdelta agonists induce the phosphorylation of Erk1/Erk2 and p38, which are known to be involved in the increase of mRNA stability. The inhibition of these MAPK signaling pathways resulted in a significant suppression of the induced IL-8 expression and the reduced mRNA stability. Therefore, our data provide the first evidence that PPARdelta induces IL-8 expression in non-stimulated endothelial cells via transcriptional as well as posttranscriptional mechanisms.

29: Clinical immunology (Orlando, Fla.), 2010 Jun 28, 72(1)
Regulation of IL-8 production by complement-activated product, C5a, in vitro and in vivo during sepsis.

[Abstract]Excessive complement-activated product complement 5a (C5a) has been implicated in the pathogenesis of sepsis development. Herein, we employed in vitro and in vivo models of sepsis to investigate the functional relationship between overtly produced C5a and IL-8. Our data revealed that C5a could strongly amplify IL-8 expression from human whole blood cells induced by LPS and other types of TLR agonists. ERK1/2 and p38, but not JNK, were mainly participated in signaling pathways for IL-8 production. In the whole blood stimulated by Escherichiacoli, C5a levels were quickly elevated and blockage of C5a significantly decreased E. coli-elicited IL-8 production. In the mouse model of sepsis induced by cecal ligation and puncture (CLP), the markedly increased keratinocyte-derived cytokine (KC) could be strongly suppressed by blockage of C5a. These data suggest that excessive C5a functions as a critical inflammatory mediator to enhance IL-8 production mainly through MAPK signaling pathways.

30: Scandinavian journal of immunology, 2010 Jul, 72(1)
Chitinase Induce the Release of IL-8 in Human Airway Epithelial Cells, Via Ca(2+)-dependent PKC and ERK Pathways.

[Abstract]Abstract Chitinases are produced in significant quantities by hosts defending against infections with chitin-containing organisms. However, little is known about the immune response of exogenous chitinase in human epithelial cells. IL-8 has been suggested to have a role in the pathogenesis of the allergenic inflammation of bronchial asthma. We examined whether Streptomyces griseus (S. griseus) chitinase-induced IL-8 on airway epithelium and identified the involvement of intracellular signalling pathways. H292 cells were treated with S. griseus chitinase with different concentrations and times. The IL-8 levels were determined by specific human IL-8 enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase polymerase chain reaction. Using a series of pharmacological inhibitors, we examined the upstream signalling pathway responsible for IL-8 expression in response to S. griseus chitinase. Cells exposed to S. griseus chitinase showed higher level of IL-8 protein production and mRNA expression. Cells stimulated by S. griseus chitinase resulted in the activation of protein kinase C (PKC), extracellular signal-regulated kinase (ERK) and nuclear factor kappa-B (NF-kB) pathways. Inhibitors of Ca(2+)-dependent PKC (Ro-31-8220, calphostin C and Go6976) significantly abolished chitinase-induced expression of IL-8. However, Ca(2+)-independent PKC inhibitor (rottlerin) did not inhibit IL-8 expression. Through ERK inhibitor (U0126) and NF-kB inhibitor (caffeine acid phenethyl ester) treatment, it was proven that ERK and NF-kB regulated chitinase-induced IL-8 expression. We concluded that S. griseus chitinase-induced IL-8 expression was regulated by the activation of Ca(2+/-)-dependent PKC, ERK and NF-kB in human airway epithelial cells.

31: Journal of neuro-oncology, 2010 Jun 25, 30(9)
IL-8 is a mediator of NF-kappaB induced invasion by gliomas.

[Abstract]Glioblastoma (GBM) is the most common and deadly form of primary brain tumor with a median survival of eleven months, despite use of extensive chemotherapy, radiotherapy and surgery. We have previously shown that nuclear factor-kappa B (NF-kappaB) is aberrantly expressed in GBM tumors and primary cell lines derived from tumor tissue. Here we show that IL-8, a chemokine is also aberrantly expressed by GBM cell lines and expression of IL-8 is in large part, attributable to the aberrant activation of NF-kappaB. We hypothesized that invasiveness of GBM cells is driven at least in part by aberrantly expressed IL-8. In support of the hypothesis we found that treatment of glioma cells with an IL-8 neutralizing antibody markedly decreased their invasiveness compared to cells treated with control IgG or left untreated. Furthermore, downregulation of IL-8 protein production with use of IL-8 targeted siRNA also resulted in decreased invasion in matrigel. We next investigated the presence of IL-8 receptors by FACS analysis and found that GBM cells (U87, U251, D54 and LN229) only express CXCR1 but not CXCR2. Treatment of U87 cells with a blocking CXCR1 antibody reduced their invasion through matrigel. Finally, we found that addition of exogenous IL-8, following downregulation of NF-kappaB which results in loss of endogenous IL-8 production, incompletely restored tumor cell invasion. Our data indicate that IL-8 is necessary but not solely responsible for glioma cell invasion and mediates its effect in an autocrine manner.

32: Journal of acquired immune deficiency syndromes (1999), 2010 Jun 23, 30(9)
IL-8 Decreases HIV-1 Transcription in Peripheral Blood Lymphocytes and Ectocervical Tissue Explants.

[Abstract]IL-8 is enhanced in the peripheral blood and lymphoid tissue of HIV-infected individuals, suggesting that IL-8 is important in the pathogenesis of HIV-1 infection and progression to AIDS. Characterizing the mechanisms of IL-8 regulation of HIV-1 replication may be relevant in addressing the role of IL-8 as a therapeutic target in HIV-1 infection. We evaluated replication of primary R5-tropic HIV-1 in peripheral blood lymphocytes and ectocervical tissue explants infected in vitro in the presence of physiological concentrations of IL-8 found in the serum and genital tract secretions of HIV-infected individuals. To identify the specific stages of the viral life cycle targeted by IL-8, we performed real-time polymerase chain reaction to detect HIV-1 reverse transcription, integration, and transcription. Early during the infection, IL-8 decreased HIV-1 reverse transcription and viral integration. This effect was transient, as on day 4 after infection, we detected no differences on HIV-1 DNA or proviral DNA in peripheral blood lymphocyte. IL-8 decreased HIV-1 transcription in both lymphocytes and ectocervical tissue explants. The decrease in viral RNA expression was associated with reduced HIV-1 replication, as measured by viral p24 release in the culture supernatants. This is the first report to suggest that IL-8 decreases replication of primary R5-tropic HIV-1 by transcriptional mechanisms.

33: Molecular endocrinology (Baltimore, Md.), 2010 Jun 23, 30(9)
A Novel Isoform of Microphthalmia-Associated Transcription Factor Inhibits IL-8 Gene Expression in Human Cervical Stromal Cells.

[Abstract]Cervical ripening during pregnancy is a profound change in cervix structure and function characterized by increases in the proinflammatory cytokine IL-8 and dissolution of the cervical extracellular matrix. Relatively little is known about the molecular mechanisms that underlie these events. Here, we report identification of a novel isoform of micropthalmia-associated transcription factor in human cervical stromal cells (MiTF-CX) that is down-regulated 12-fold during cervical ripening and that represses expression of IL-8. Ectopic expression of MiTF-CX in human cervical stromal cells resulted in substantial suppression of endogenous IL-8 mRNA and protein expression, whereas expression of dominant negative MiTF-CX mutants with impaired DNA binding resulted in dramatic increases in IL-8 production. Gel shift, reporter gene, and chromatin immunoprecipitation assays revealed one strong binding site (E-box (-397) CACATG(-391)) in the human IL-8 promoter that was crucial for mediating transcriptional repression by MiTF-CX. Moreover, we show that MiTF-CX expression in the cervix was itself positively autoregulated via two E-box motifs within a 2.1-kb promoter fragment. We therefore propose that maintenance of cervical competency during pregnancy is an active process maintained through suppression of IL-8 by the transcription factor MiTF-CX. During cervical ripening, loss of MiTF-CX would result in significant up-regulation of IL-8 mRNA and protein synthesis, thereby leading to recruitment and activation of leukocytes within the cervix and dissolution of the extracellular matrix.

34: Schizophrenia research, 2010 Jun 6, 10(1)
Structural brain alterations in schizophrenia following fetal exposure to the inflammatory cytokine interleukin-8.

[Abstract]BACKGROUND: Maternal infection during pregnancy has been repeatedly associated with increased risk for schizophrenia. Nevertheless, most viruses do not cross the placenta; therefore, the damaging effects to the fetus appear to be related to maternal antiviral responses to infection (e.g. proinflammatory cytokines). Fetal exposure to the proinflammatory cytokine interleukin-8 (IL-8) has been significantly associated with risk of schizophrenia in offspring. This study sought to determine the association between fetal exposure to IL-8 and structural brain changes among schizophrenia cases and controls. METHODS: Subjects were 17 cases diagnosed with schizophrenia from the Developmental Insult and Brain Anomaly in Schizophrenia (DIBS) study. Psychiatric diagnoses were determined among offspring with semi-structured interviews and medical records review. IL-8 was determined from assays in archived prenatal sera and volumetric analyses of neuroanatomical regions were obtained from T1-weighted magnetic resonance imaging in adulthood. Eight controls were included for exploratory purposes. RESULTS: Among cases, fetal exposure to increases in IL-8 was associated with significant increases in ventricular cerebrospinal fluid, significant decreases in left entorhinal cortex volumes and significant decreases in right posterior cingulate volumes. Decreases that approached significance also were found in volumes of the right caudate, the putamen (bilaterally), and the right superior temporal gyrus. No significant associations were observed among controls. CONCLUSION: Fetal exposure to elevations in maternal IL-8 led to structural neuroanatomic alterations among cases in regions of the brain consistently implicated in schizophrenia research. In utero exposure to elevations in IL-8 may partially account for brain disturbances commonly found in schizophrenia.

35: International archives of medicine, 2010 Jun 15, 3(1)
Interleukin 8 (IL-8) - a universal biomarker?

[Abstract]ABSTRACT: Many clinical conditions including various types of cancers are complex and generally require invasive, laborious, expensive and time-consuming investigations for their diagnosis, treatment and follow-up. There is therefore a general need for exploring non-invasive markers in clinical medicine. Interleukin 8 (IL-8) is currently being applied in various sub specialties of medicine either for the purpose of rapid diagnosis or as a predictor of prognosis. Nevertheless, there is need for large-scale studies to substantiate accuracy and outcome. This article will summarize current evidence suggesting that Interleukin 8 (IL-8) may serve as a useful biomarker.

36: BMC microbiology, 2010 Jun 14, 10(1)
LPS-induced IL-8 activation in human intestinal epithelial cells is accompanied by specific histone H3 acetylation and methylation changes.

[Abstract]ABSTRACT: BACKGROUND: The release of LPS by bacteria stimulates both immune and specific epithelial cell types to release inflammatory mediators. It is known that LPS induces the release of IL-8 by intestinal mucosal cells. Because it is now emerging that bacteria may induce alteration of epigenetic patterns in host cells, we have investigated whether LPS-induced IL-8 activation in human intestinal epithelial cells involves changes of histone modifications and/or DNA methylation at IL-8 gene regulatory region. RESULTS: RT-PCR analysis showed that IL-8 mRNA levels rapidly increase after exposure of HT-29 cells to LPS. DNA demethylating agents had no effects on IL-8 expression, suggesting that DNA methylation was not involved in IL-8 gene regulation. Consistently we found that 5 CpG sites located around IL-8 transcription start site (-83, -7, +73, +119, +191) were unmethylated on both lower and upper strand either in LPS treated or in untreated HT-29 cells, as well as in normal intestinal mucosa. Conversely, pretreatment of HT-29 cells with deacetylase inhibitors strengthened the LPS-mediated IL-8 activation. Inhibitors of histone deacetylases could induce IL-8 mRNA expression also in the absence of LPS, suggesting that chromatin modifications could be involved in IL-8 gene regulation. Chromatin immunoprecipitation analyses showed that, concurrently with IL-8 activation, transient specific changes in H3 acetylation and H3K4, H3K9 and H3K27 methylation occurred at IL-8 gene promoter during LPS stimulation. Changes of H3-acetyl, H3K4me2 and H3K9me2 levels occurred early, transiently and corresponded to transcriptional activity, while changes of H3K27me3 levels at IL-8 gene occurred later and were long lasting. CONCLUSIONS: The results showed that specific chromatin modifications occurring at IL-8 gene, including histone H3 acetylation and methylation, mark LPS-mediated IL-8 activation in intestinal epithelial cells while it is unlikely that DNA methylation of IL-8 promoter is directly involved in IL-8 gene regulation in these cells.

37: Toxicology in vitro : an international journal published in association with BIBRA, 2010 Jun 8, 10(1)
Use of IL-8 release and p38 MAPK activation in THP-1 cells to identify allergens and to assess their potency in vitro.

[Abstract]The local lymph node assay (LLNA) has been developed to assess skin sensitization, and based on the EC3 value, it can also be used to evaluate allergen potency. Therefore, in the development of in vitro alternatives to the LLNA assay, one should not only consider the hazard identification but also the possibility to classify allergens relatively to their potency. We have recently described a selective release of interleukin-8 (IL-8) by chemical allergens in THP-1 cell line, and identified the activation of p38 mitogen-activated protein kinase (p38 MAPK) as a common pathway. Therefore, the purpose of this study was to expand the number of chemicals tested and to investigate whether IL-8 production and p38 MAPK activation can be used to classify allergens according to their potency. THP-1 cells were exposed to the contact allergens (p-benzoquinone, 2-aminophenol, isoeugenol, diethyl maleate, citral and imidazolidinyl urea), selected according to their potency in the LLNA, and to lactic acid and propylene glycol as non-sensitizers. p38 MAPK activation was evaluated 5-15 min after treatment by FACS analysis, while IL-8 release was assed by ELISA following 24 h of incubation. p38 MAPK was activated by all contact allergens, including the pro-apten isoeugenol, whereas IL-8 release was significantly increased after stimulation with all allergens tested, except for isoeugenol. The failure of isoeugenol may be due to decrease in the stability of IL-8 mRNA. Irritants exposure, as expected, failed to induce both p38 MAPK activation and IL-8 release. A significant correlation between IL-8 release and the LLNA EC(3) was found (Pearson correlation r= 0.743, p= 0.0036, n=12). On the contrary, the activation of p38 MAPK showed no significant correlation between LLNA data and vigor of p38 MAPK activation. Overall, data presented confirm our previous observations and reveal IL-8 as potential tool not only to identify sensitizers, with the exception of pro-haptens, but also to classify them according to their potency, while p38 MAPK activation allows the identification of all sensitizers, including pro-haptens, but was not useful for potency classification.

38: Virus research, 2010 Jun 9, 10(1)
Respiratory echovirus 30 and coxsackievirus B5 can induce production of RANTES, MCP-1 and IL-8 by human bronchial epithelial cells.

[Abstract]Human Enteroviruses (HEV) (picornaviridae) are considered as one the major viral causes of childhood acute respiratory wheezing illnesses including bronchiolitis and asthma exacerbation. To identify the mechanisms that can regulate the development of airway mucosa inflammation during HEV respiratory lower tract infection, we investigated the profile and the levels of mRNA and protein secretion for CC and CXC human chemokines by HEV-infected primary human bronchial epithelial cells (SAE cells) using RT-PCR array and Bio-Plex assay. Cultures of SAE cells were infected by reference and wild-type HEV respiratory strains, demonstrating a replicative and productive viral infection. We observed that the replicative infection of the SAE cells by reference and wild-type HEV strains induced specific dose and time-dependent increases in mRNA and protein secretion only for RANTES, MCP-1 and IL-8 and not for all other CC and CXC human chemokines tested. The protein secretion of these chemokines appeared to be significantly increased at 48 or 72h post-infection in cultures treated by low-doses of IFN-gamma comparatively to mock-infected cells (P<0.001), and was correlated to the viral replication activity. In conclusion, our findings demonstrated a selective production of RANTES, IL-8 and MCP-1 released by HEV-infected epithelial cells of the small bronchioles along with mechanisms of amplification mediated by IFN-gamma.

39: Retina (Philadelphia, Pa.), 2010 Jun 3, 115(22)
AQUEOUS HUMOR AND PLASMA LEVELS OF VASCULAR ENDOTHELIAL GROWTH FACTOR AND INTERLEUKIN-8 IN PATIENTS WITH CENTRAL SEROUS CHORIORETINOPATHY.

[Abstract]PURPOSE:: The purpose of this study was to determine aqueous vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) levels in patients with central serous chori-oretinopathy (CSC) before a single intravitreal bevacizumab injection. METHODS:: Twelve eyes with symptomatic CSC were included. Samples from patients with cataracts served as controls. The levels of VEGF and IL-8 concentrations were measured in aqueous humor and plasma by multiplex bead assays. RESULTS:: All patients with CSC showed an improvement in visual acuity and resolved neurosensory detachment after intravitreal bevacizumab injection. The aqueous humor levels of VEGF and IL-8 were not significantly increased in patients with CSC compared with the healthy control group (18.2 +/- 24.8 vs. 35.3 +/- 28.5 pg/mL, P > 0.05; 2.3 +/- 0.4 vs. 2.8 +/- 0.3 pg/mL, P > 0.05, respectively). The plasma levels of VEGF and IL-8 in patients with CSC were not different from those in the healthy control group. CONCLUSION:: Vascular endothelial growth factor and IL-8 were not increased in the aqueous humor and plasma of patients with CSC. The effect of intravitreal bevacizumab injection as a treatment for CSC must be fully understood, and the true effect of anti-VEGF treatment in patients with CSC remains to be elucidated.

40: Immunology, 2010 May 26, 40(6)
The role of endothelial interleukin-8/NADPH oxidase 1 axis in sepsis.

[Abstract]Summary Sepsis is a generalized inflammatory disease, caused by the hyperinflammatory response of the host, rather than by invading organisms. Endothelial cells play a crucial role in the pathogenesis of sepsis. In this study, we investigated the effects of interleukin-8 (IL-8), a known neutrophil chemoattractant, on lipopolysaccharide (LPS) -induced reactive oxygen species (ROS) production by endothelial cells, and its significance in the pathogenesis of LPS-mediated sepsis. The results revealed that IL-8 directly induced ROS production in human umbilical vein endothelial cells (HUVECs), and also mediated LPS-induced ROS production by HUVECs. Stimulation of HUVECs by LPS strongly enhanced tissue factor expression, a hallmark of severe sepsis, which was suppressed by IL-8 knockdown. We further discovered that NADPH oxidase (Nox) 1 expression in LPS-stimulated HUVECs was markedly repressed by IL-8 knockdown, and Nox1 knockdown reduced tissue factor expression, suggesting that the LPS/IL-8 signalling in endothelial cells was predominantly mediated by Nox1. In conclusion, LPS stimulation of endothelial cells causes activation of the IL-8-Nox1 axis, enhances the production of ROS, and ultimately contributes to the progression of severe sepsis.

41: Infection and immunity, 2010 Jun 1, 24(6)
Differential expression of IL-8 by PMNs of two closely related species, Ovis canadensis and Ovis aries, in response to Mannheimia haemolytica infection.

[Abstract]Pneumonic lesions and mortality caused by Mannheimia haemolytica in bighorn sheep (Ovis canadensis, BHS) are more severe than those in the related species, domestic sheep (Ovis aries, DS), both under natural and experimental conditions. Leukotoxin (Lkt) and lipopolysaccharide (LPS) are the most important virulence factors of this organism. One hallmark of pathogenesis of pneumonia is the influx of polymorphonuclear leukocytes (PMNs) into the lungs. Lkt-induced cytolysis of PMNs results in the release of cytotoxic compounds capable of damaging lung tissue. Interleukin-8 (IL-8) is a potent PMN chemoattractant. The objective of this study was to determine if there is differential expression of IL-8 by macrophages and PMNs of BHS and DS in response to M. haemolytica. Macrophages and PMNs of BHS and DS were stimulated with heat-killed M. haemolytica or LPS. IL-8 expression by the cells was measured by enzyme-linked immunosorbent assays and real-time RT-PCR. PMNs of BHS expressed several fold higher levels of IL-8 than those of DS upon stimulation. Lesional lung tissue of M. haemolytica-infected BHS contained significantly higher levels of IL-8 than nonlesional tissue. Bronchoalveolar lavage (BAL) fluid of infected BHS also contained higher levels of IL-8 than that of infected DS. Depletion of IL-8 reduced migration of PMNs toward BAL fluid by approximately 50%, indicating that IL-8 is integral to PMN recruitment to the lung during M. haemolytica infection. Excessive production of IL-8, enhanced recruitment of PMNs and their lysis by Lkt are likely responsible for the severity of lung lesions in M. haemolytica-infected BHS.

42: Molecular nutrition & food research, 2010 May 28, 40(6)
Amino acids stimulate Akt phosphorylation, and reduce IL-8 production and NF-kappaB activity in HepG2 liver cells.

[Abstract]Hepatic insulin resistance and inflammatory cytokine production contribute to the manifestation of the metabolic syndrome. As amino acids have been implicated in modulating insulin signaling and inflammation, we have investigated the effects of glutamine, leucine and proline on markers of inflammation and insulin sensitivity in HepG2 liver cells. Cells were incubated with IL-1beta (5 ng/mL) to stimulate IL-8 production. After 24 h, glutamine inhibited IL-8 production significantly (p<0.05) at 2, 5 and 10 mM (to 82, 73 and 72% of control), whereas leucine reduced IL-8 production significantly only at 10 mM (66%) and proline at 5 and 10 mM (71 and 52%). Glutamine, leucine and proline all reduced NF-kappaB activity after 3 h of IL-1beta stimulation at 2, 5 and 10 mM (p<0.001). Insulin-induced (1 nM) Akt phosphorylation was reduced in cells treated with tumour necrosis factor-alpha (10 ng/mL) for 24 h, but was partly restored by simultaneous incubation with glutamine, leucine and proline (25 mM). Phosphorylation of glycogen synthase kinase-3beta was unaffected by insulin stimulation and amino acid treatment. Our results indicate that glutamine, leucine and proline attenuate IL-8 production, probably through inhibition of NF-kappaB, and that they increase Akt phosphorylation in HepG2 cells.

43: Biochemical and biophysical research communications, 2010 May 25, 40(6)
Thromboxane A(2) increases endothelial permeability through upregulation of interleukin-8.

[Abstract]Thromboxane A(2) (TXA(2)), a major prostanoid formed from prostaglandin H(2) by thromboxane synthase, is involved in the pathogenesis of a variety of vascular diseases. In this study, we report that TXA(2) mimetic U46619 significantly increases the endothelial permeability both in vitro and in vivo. U46619 enhanced the expression and secretion of interleukin-8 (IL-8), a major inducer of vascular permeability, in endothelial cells. Promoter analysis showed that the U46619-induced expression of IL-8 was mainly regulated by nuclear factor-kappaB (NF-kappaB). U46619 induced the activation of NF-kappaB through IkappaB kinase (IKK) activation, IkappaB phosphorylation and NF-kappaB nuclear translocation. Furthermore, the inhibition of IL-8 or blockade of the IL-8 receptor attenuated the U46619-induced endothelial cell permeability by modulating the cell-cell junctions. Overall, these results suggest that U46619 promotes vascular permeability through the production of IL-8 via NF-kappaB activation in endothelial cells.

44: Planta medica, 2010 May 26, 40(6)
Green and Black Tea Inhibit Cytokine-Induced Il-8 Production and Secretion in AGS Gastric Cancer Cells via Inhibition of NF-kappaB Activity.

[Abstract]Consumption of tea is associated with a reduced risk for several gastrointestinal cancers. Inflammatory processes, such as secretion of IL-8 from the gastric epithelium in response to chronic chemokine or antigen exposure, serve both as a chemoattractant for white blood cells and a prerequisite for gastric carcinogenesis. In this study, the gastric adenocarcinoma cell line AGS was used to investigate the effect of green tea extract, black tea extract, and epigallocatechin gallate (EGCG), the most abundant catechin in tea, on cytokine-induced inflammation. AGS cells were stimulated with interleukin-1 beta (IL-1 beta) to initiate inflammation, followed by exposure to either tea extracts or EGCG. We found that both green and black tea extracts at concentrations of 20 and 2 microM total catechins, respectively, significantly (p < 0.05) inhibited IL-1 beta-induced IL-8 production and secretion to a similar extent. Treatment of AGS cells with EGCG (8 microM) produced similar reductions in IL-1 beta-induced IL-8 production and secretion. Inhibition of NF- kappaB activity was found to be responsible, in part, for these observed effects. Our findings demonstrate that both green and black tea extracts with distinctly different catechin profiles, are capable of disrupting the molecular link between inflammation and carcinogenesis via inhibition of NF- kappaB activity in AGS cells.

45: Scandinavian journal of immunology, 2010 Jun, 71(6)
IL-8 in cerebrospinal fluid from children with acute encephalopathy is higher than in that from children with febrile seizure.

[Abstract]We identify possible differences in the cytokine/chemokine profiles in cerebrospinal fluid (CSF) from children with encephalopathy and febrile seizure. Interleukin (IL)-1beta, 2, 4, 5, 6, 7, 8, 10, 12, 13, 17, interferon-gamma, tumour necrosis factor-alpha, granulocyte colony-stimulating factor, granulocyte monocyte colony-stimulating factor, monocyte chemoattractant protein-1 and macrophage inflammatory protein-1beta were measured simultaneously in CSF supernatants from children with encephalopathy (n = 8), febrile seizure (n = 16) and fever without neurological complications (n = 8). IL-8 in CSF from children with encephalopathy was significantly elevated compared to that in CSF from children with febrile seizure and fever without neurological complications. IL-8 in CSF was also higher than serum IL-8, suggesting that increased IL-8 was generated from glia cells or astrocytes, not by leakage from serum. Increased IL-8 in CSF in encephalopathy may protect against severe brain damage.

46: Surgery today, 2010 Jun, 40(6)
The role of serum interleukin-8 in hepatic resections.

[Abstract]PURPOSE: Interleukin-8 (IL-8) is a neutrophil chemotactic factor, which is associated with some inflammatory diseases and various types of surgical stress. The aim of this study was to investigate whether the early postoperative serum IL-8 level may potentially be a new indicator of a surgical stress in patients undergoing a hepatic resection. METHODS: The serum IL-8 levels were measured in 37 patients who underwent a hepatectomy. The serum IL-8 levels were serially measured using an enzyme-linked immunosorbent assay both before and after a hepatic resection. In addition, the correlation between the postoperative IL-8 value and several clinical variables were examined. RESULTS: The mean level of IL-8 significantly increased immediately after the operation (P < 0.01 vs before the operation) and decreased on the first postoperative day (POD 1, P < 0.05 vs after the operation). The early postoperative IL-8 levels positively correlated with the length of the procedure (r = 0.383; P < 0.05), the estimated blood loss (r = 0.483; P < 0.01) and the serum bilirubin level on POD 1 (r = 0.390; P < 0.05), and inversely correlated with the white blood cell counts (r = -0.388; P < 0.05) and lymphocyte counts on POD 1 (r = -0.424; P < 0.05). In a comparison of the postoperative IL-8 levels with the surgical factors, there was a significant difference in the extension of the resection (P < 0.05) and in blood transfusion. The patients with a fever of more than 38 degrees C showed higher levels of IL-8 immediately after the operation than those without fever (P < 0.01). CONCLUSIONS: The early postoperative serum IL-8 level was found to correlate with the degree of the severity of surgery in patients undergoing a hepatic resection, and it is also considered to be a new indicator of surgical stress and liver injury.

47: Journal of dairy science, 2010 Jun, 93(6)
Effect of nonfat dry milk and major whey components on interleukin-6 and interleukin-8 production in human intestinal epithelial-like Caco-2 cells.

[Abstract]Bovine nonfat dry milk (NDM) and major whey components (lactose, alpha-lactalbumin, and beta-lactoglobulin) were evaluated for their effects on IL-6 and IL-8 production in human intestinal-like Caco-2 cells unstimulated or stimulated with IL-1beta. All the whey components investigated and NDM induced IL-6 production by Caco-2 cells; the most significant increase was observed with beta-lactoglobulin. In the case of IL-1beta-stimulated cells, neither NDM nor the major whey components investigated contributed to the induction of IL-6 production after they were stimulated. Induction of IL-8 production by both alpha-lactalbumin and beta-lactoglobulin was higher than that by lactose and NDM; alpha-lactalbumin was a more potent inducer of IL-8 than beta-lactoglobulin and IL-1beta alone in both unstimulated and stimulated cells. In Caco-2 cells that were stimulated with IL1-beta, NDM and all the major whey components investigated had a synergistic effect on induction of IL-8 production, indicating that IL-8 induction was amplified by prior stimulation of cells by IL-1beta. This synergistic effect was not observed with IL-6. Our results suggest that immunomodulatory properties of milk components may be affected by other complex events in the gut.

48: Clinica chimica acta; international journal of clinical chemistry, 2010 May 17, 195(1)
Association of haplotypes in the IL8 gene with susceptibility to chronic periodontitis in a Brazilian population.

[Abstract]BACKGROUND: Interleukin 8 (IL-8) is a chemokine related to the initiation and amplification of acute and chronic inflammatory processes. Polymorphisms in the IL8 gene have been associated with inflammatory diseases. We investigated whether the -845(T/C) and -738(T/A) single nucleotide polymorphisms (SNPs) in the IL8 gene, as well as the haplotypes they form together with the previously investigated -353(A/T), are associated with susceptibility to chronic periodontitis. METHODS: DNA was extracted from buccal epithelial cells of 400 Brazilian individuals (control n=182, periodontitis n=218). SNPs were genotyped by the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method. Disease associations were analyzed by the chi(2) test, Exact Fisher test and Clump program. Haplotypes were reconstructed using the expectation-maximization algorithm and differences in haplotype distribution between the groups were analyzed to estimate genetic susceptibility for chronic periodontitis development. RESULTS: When analyzed individually, no SNPs showed different distributions between the control and chronic periodontitis groups. Although, nonsmokers carrying the TTA/CAT (OR=2.35, 95% CI=1.03-5.36) and TAT/CTA (OR=6.05, 95% CI=1.32-27.7) haplotypes were genetically susceptible to chronic periodontitis. The TTT/TAA haplotype was associated with protection against the development of periodontitis (for nonsmokers OR=0.22, 95% CI=0.10-0.46). CONCLUSION: Although none of the investigated SNPs in the IL8 gene were individually associated with periodontitis, some haplotypes showed significant association with susceptibility to, or protection against, chronic periodontitis in a Brazilian population.

49: Pediatrics international : official journal of the Japan Pediatric Society, 2010 May 6, 185(3)
Elastase, alpha-proteinase inhibitor, and interleukin-8 in pre-dialyzed and hemodialyzed patients with chronic kidney disease.

[Abstract]Abstract Background: Neutrophil elastase in complex with alpha(1)-proteinase inhibitor (NE-alpha(1)PI) and IL-8 may serve as indicators of neutrophil activation and inflammatory stage. The aim of the study was to evaluate NE-alpha(1)PI, alpha(1)-PI, and IL-8 levels in the blood of patients with chronic kidney disease (CKD) undergoing hemodialysis (HD) or treated conservatively (CT). The influence of a single HD session on the investigated parameters also was assessed. Material and methods: Blood samples were obtained from two groups of hemodialyzed patients, i.e. children/young adults (group HD1, n = 8) and adults (HD2, n = 13), as well as 13 CT patients and a group of healthy subjects (C). The proteins were measured by ELISA or radial immunodiffusion. Results: There were no significant differences in NE-alpha(1)PI, alpha(1)-PI, and IL-8 concentrations between the HD1 and HD2 patients. The levels of NE-alpha(1)PI were considerably higher than normal in both groups of HD patients (before and after the HD session) and in the CT patients. Higher titers of NE-alpha(1)PI (P < 0.05) and alpha(1)-PI (P < 0.01) were obtained in the adults during the course of HD. Increased NE-alpha(1)PI positively correlated with alpha(1)-PI. The serum concentration of IL-8 was significantly higher in the HD2 patients before and after dialysis than in the controls. Conclusions: The data indicate that in CKD patients neutrophils are highly activated both in the pre-dialyzed period and on regular hemodialysis. Contact with the dialysis membrane during HD causes a significant increase in blood NE-alpha(1)PI and alpha(1)-PI in adults, but not in children /young adults. NE-alpha(1)PI seems to be a much better indicator of an inflammatory state in CKD patients than free alpha(1)-PI and IL-8.

50: Clinical immunology (Orlando, Fla.), 2010 May 17, 195(1)
Short-term IL-1beta blockade reduces monocyte CD11b integrin expression in an IL-8 dependent fashion in patients with type 1 diabetes.

[Abstract]OBJECTIVE: Interleukin 1-beta (IL-1beta) is a major inflammatory cytokine. Blockade of the IL-1beta pathway is therapeutically efficacious in type 2 diabetes, but the mechanistic effects on the immune system are incompletely understood. RESEARCH DESIGN: We administered an IL-1 receptor antagonist, anakinra, to 7 type 1 diabetes patients in order to investigate the immunologic and metabolic effects of this drug. Mechanistic assays were performed before and after drug administration. RESULTS: A novel signature was observed, with reduced serum interleukin 8 (IL-8) levels and reduced CD11b integrin expression on monocytes associated with increased CXCR1 expression. CONCLUSIONS: This set of linked phenotypes suggests that blockade of the IL-1beta pathway results in the reduced ability of mononuclear cells to traffic to sites of inflammation. Mechanistic studies from large scale trials using IL-1 blockade in type 1 diabetes should focus on changes in monocyte trafficking and the IL-8 pathway.

51: Placenta, 2010 May 17, 195(1)
Role of Interleukin 8 in Uterine Natural Killer Cell Regulation of Extravillous Trophoblast Cell Invasion.

[Abstract]BACKGROUND: Extravillous trophoblast cell (EVT) invasion of maternal tissues is critical for successful pregnancy. Decidual factors, including uterine natural killer (uNK) and T cell derived cytokines play a role in regulating this process. Interleukin (IL) 8 has been implicated as a regulator of EVT invasion. HYPOTHESIS: uNK cell stimulation of EVT invasion is associated with IL-8 levels. METHODS: CD8+, total decidual and CD56(+) uNK cells (8-10 and 12-14 weeks gestational age) were cultured. IL-8 mRNA and protein levels were determined. IL-8 receptors (IL-8RA and IL-8RB) were localised in first trimester placental bed biopsies. The effect of IL-8 +/- IL-8 neutralising antibodies and CD8+ T cell or uNK cell supernatants +/- IL-8 neutralising antibodies on EVT invasion was assessed. EVT secreted levels of MMP-2, MMP-9, uPA, PAI-1 and PAI-2 were assessed by substrate zymography or Western Blot. RESULTS: High levels of IL-8 protein and mRNA were detected in all samples. IL-8RA and IL-8RB were expressed by EVT. Exogenous IL-8 stimulated EVT invasion in a paracrine manner. uNK cell supernatants, but not CD8+ cell supernatants, stimulated EVT invasion. IL-8 neutralising antibody partially abrogated this uNK cell stimulated invasion. IL-8 increased levels of secreted MMP-2, but did not alter any of the other proteases or protease inhibitors tested. CONCLUSION: uNK cell stimulation of EVT invasion is partially mediated by IL-8. Unstimulated CD8+ T cells do not alter EVT invasion despite secreting similar levels of IL-8 as uNK cells.

52: Journal of neuro-oncology, 2010 May 9, 185(3)
Knockdown of CypA inhibits interleukin-8 (IL-8) and IL-8-mediated proliferation and tumor growth of glioblastoma cells through down-regulated NF-kappaB.

[Abstract]Although cyclophilin A (CypA) has been reported to be over-expressed in cancer cells and solid tumors, its expression and role in glioblastomas have not been studied. Herein, we show that expression of CypA in human glioblastoma cell lines and tissues is significantly higher than in normal human astrocytes and normal counterparts of brain tissue. To determine the role of over-expressed CypA in glioblastoma, stable RNA interference (RNAi)-mediated knockdown of CypA (CypA KD) was performed in gliobastoma cell line U87vIII (U87MG . DeltaEGFR). CypA KD stable single clones decrease proliferation, infiltration, migration, and anchorage-independent growth in vitro and with slower growth in vivo as xenografts in immunodeficient nude mice. We have also observed that knockdown of CypA inhibits expression of interleukin-8 (IL-8), a tumorigenic and proangiogenic cytokine. Conversely, enforced expression of CypA in the CypA KD cell line, Ud-12, markedly enhanced IL-8 transcripts and restored Ud-12 proliferation, suggesting that CypA-mediated IL-8 production provides a growth advantage to glioblastoma cells. CypA knockdown-mediated inhibition of IL-8 is due to reduced activity of NF-kappaB, which is one of the major transcription factors regulating IL-8 expression. These results not only establish the relevance of CypA to glioblastoma growth in vitro and in vivo, but also suggest that small interfering RNA-based CypA knockdown could be an effective therapeutic approach against glioblastomas.

53: Journal of periodontology, 2010 May 3, 185(3)
DNA methylation status of the IL8 gene promoter in aggressive periodontitis.

[Abstract]Background: Studies evaluating the methylation status of cytokines genes may have relevance for inflammatory diseases in which the expression of some cytokines is altered, such as periodontitis. The aim of the present study was to observe the DNA methylation status in the Interleukin 8 (IL8) gene promoter in cells of the oral epithelium of subjects affected by generalized aggressive periodontitis (AgP) and to compare it with those of control subjects. Methods: Genomic DNA from epithelial oral cells of 37 generalized AgP patients and 37 controls were purified and modified by sodium bisulphite. Modified DNA was submitted by methylation-specific polymerase chain reaction (MSP), electrophoresed on 10% polyacrylamide gels and stained with SYBR Gold. Results: Subjects who presented generalized AgP have a higher frequency of hypomethylation of the IL8 gene promoter in oral epithelium cells than those controls (86.5% in the generalized AgP group versus 62% in the control group; P = 0.016; chi(2) test). Conclusion: It was found a marked hypomethylated status in the oral epithelial cells of the subjects who presented generalized AgP, comparing to the controls, in the promoter region of the IL8 gene. This hypomethylated status may reflect a generalized condition of oral epithelial cells, including gingival epithelium, since gingival epithelial cells were also collected during mouth wash.

54: ChemMedChem, 2010 May 5, 185(3)
Metronidazole-Deoxybenzoin Derivatives as Anti-Helicobacter pylori Agents with Potent Inhibitory Activity against HPE-Induced Interleukin-8.

[Abstract]A series of new metronidazole-deoxybenzoin derivatives were synthesized and evaluated for their antimicrobial activity against Helicobacter pylori. Highly selective anti-H. pylori activity was also observed in synthesized compounds. Compound 34 exhibited the most potent activity, similar to the positive control amoxicillin. Furthermore, compounds 17 and 34 were able to significantly decrease H. pylori water extract (HPE)-induced production of interleukin-8 (IL-8) in gastric mucosal cells, which did not show any effect on the cell viability.

55: The Journal of biological chemistry, 2010 May 4, 185(3)
Generation of cyclic ADP-Ribose and nicotinic acid adenine dinucleotide phosphate by CD38 for Ca2+ signaling in interleukin-8-treated lymphokine-activated killer cells.

[Abstract]We have previously demonstrated that cyclic ADP-ribose (cADPR) is a calcium signaling messenger in interleukin 8 (IL8)-induced lymphokine-activated killer (LAK) cells. In this study, we examined the possibility that IL8 activates CD38 to produce another messenger, nicotinic acid adenine dinucleotide phosphate (NAADP), in LAK cells, and we showed that IL8 induced NAADP formation following cADPR production. These calcium signaling messengers were not produced when LAK cells prepared from CD38 knockout mice were treated with IL8, indicating that the synthesis of both NAADP and cADPR is catalyzed by CD38 in LAK cells. Application of cADPR to LAK cells induced NAADP production, whereas NAADP failed to increase intracellular cADPR level, confirming that the production of cADPR precedes that of NAADP in IL8-treated LAK cells. Moreover, NAADP increased intracellular Ca2+ signaling as well as cell migration, which was completely blocked by bafilomycin A1, suggesting that NAADP is generated in lysosome-related organelles following cADPR production. IL8 or exogenous cADPR, but not NAADP, increased intracellular cAMP level. cGMP analog, 8-pCPT-cGMP, 8-(4-chlorophenylthio)-guanosine 3, 5-cyclic monophosphate, increased both cADPR and NAADP production whereas cAMP analog, 8-pCPT-cAMP, 8-(4-chlorophenylthio)-adenosine-3, 5-cyclic monophosphate, increased only NAADP production, suggesting that cAMP is essential for IL8-induced NAADP formation. Furthermore, activation of Rap1, a downstream molecule of Epac, was required for IL8-induced NAADP formation in LAK cells. Taken together, our data suggest that IL8-induced NAADP production is mediated by CD38 activation through the actions of cAMP/Epac/PKA/Rap1 in LAK cells, and that NAADP plays a key role in Ca2+ signaling of IL8-induced LAK cell migration.

56: Infection and immunity, 2010 May 3, 185(3)
Shiga toxin 2 and flagellin from Shiga toxigenic Escherichia coli super-induce IL-8 through synergistic effects on host SAPKinase activation.

[Abstract]Shiga toxins expressed in the intestinal lumen during infection with Shiga toxigenic Escherichia coli must translocate across the epithelium and enter the systemic circulation to cause systemic (pathological) effects, including haemolytic uraemic syndrome. The trans-epithelial migration of polymorphonuclear leukocytes in response to chemokine expression by intestinal epithelial cells is thought to promote uptake of Stx from the intestinal lumen by compromising the epithelial barrier. In the present study, we investigate the hypothesis that flagellin acts in conjuction with Shiga toxin to augment this chemokine expression. We investigate the relative contributions of nuclear factor-kappaB and mitogen activated protein kinase signalling to transcription and translation of interleukin-8. Using reporter gene constructs, we show that flagellin-mediated interleukin-8 gene transcription is heavily dependent on both NF-kappaB and ERK1/2 activation. In contrast, inhibition of p38 has no detectable effect on interleukin-8 gene transcription, even though flagellin-mediated activation of host p38 is critical for maximal interleukin-8 protein expression. Inhibition of MAP kinase-interacting kinase 1 suggests that p38 signalling affects the post-transcriptional regulation of interleukin-8 protein expression induced by flagellin. Co-treatment with Stx2 and flagellin results in both a synergistic up-regulation of JNKs, p38 activation, and a super-induction of interleukin-8 mRNA. This synergism was also evident at the protein level, with increased interleukin-8 protein detectable following co-treatment with flagellin and Stx2. We propose that flagellin, in conjunction with Shiga toxin, synergistically up-regulates stress-activated protein kinases, resulting in super-induction of interleukin-8, and ultimately, absorption of Stx into the systemic circulation.

57: FEBS letters, 2010 Jul 2, 584(13)
Rho-kinase mediates lysophosphatidic acid-induced IL-8 and MCP-1 production via p38 and JNK pathways in human endothelial cells.

[Abstract]Lysophosphatidic acid (LPA), an inflammatory mediator that is elevated in multiple inflammatory diseases, is a potent activator of Rho kinase (ROCK) signaling and of chemokine production in endothelial cells. In this study, LPA activated ROCK, p38, JNK and NF-kappaB pathways and induced interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) mRNA and protein expression in human endothelial cells. We mapped signaling events downstream of ROCK, driving chemokine production. In summary, MCP-1 production was partly regulated by ROCK acting upstream of p38 and JNK and mediated downstream by NF-kappaB. IL-8 production was largely driven by ROCK through p38 and JNK activation, but with no involvement of NF-kappaB.

58: Cytotechnology, 2010 Apr 28, 25(5)
Granzyme A and thrombin differentially promote the release of interleukin-8 from alveolar epithelial A549 cells.

[Abstract]Some of extracellular serine proteases with trypsin-like specificity of cleavage have been known to increase the release of inflammatory mediators from various cell types. For instance, two well-known trypsin-like serine proteases circulating in blood, granzyme A (GrA) and thrombin, have been found to promote interleukin (IL)-8 release from an alveolar epithelial A549 cell line. However, the mechanisms by which the proteases promote IL-8 release from the cells are not fully understood. In the present study, using A549 cells we found that (1) thrombin promoted IL-8 release from the cells via a mechanism partially involving activation of protease-activated receptor-1, a G-protein coupled receptor, whereas a recombinant form of GrA (rGrA) did it via a mechanism that does not involve the receptor activation; that (2) unlike rGrA, thrombin did not cause detachment and microtubule disruption of the cells; and that (3) the release of IL-8 induced by rGrA was inhibited in the presence of taxol, a microtubule-stabilizing reagent, whereas that induced by thrombin was not. These findings suggest that rGrA and thrombin promote the release of IL-8 from A549 cells through distinct mechanisms.

59: Journal of periodontal research, 2010 Apr 19, 25(5)
Oxidized low-density lipoprotein increases interleukin-8 production in human gingival epithelial cell line Ca9-22.

[Abstract]Suzuki K, Sakiyama Y, Usui M, Obama T, Kato R, Itabe H, Yamamoto M. Oxidized low-density lipoprotein increases interleukin-8 production in human gingival epithelial cell line Ca9-22. J Periodont Res 2010; doi: 10.1111/j.1600-0765.2009.01263.x. (c) 2010 John Wiley & Sons A/S Background and Objective: Recent epidemiological studies have shown a correlation between periodontitis and hyperlipidemia. We have found high levels of oxidized low-density lipoprotein (OxLDL) in the gingival crevicular fluid of dental patients. In the present study, we tried to examine the possible role of OxLDL in periodontal inflammation in vitro. Material and Methods: Cells of the human gingival epithelial cell line Ca9-22 were cultured in media containing OxLDL, and the amounts of interleukin-8 (IL-8) and prostaglandin E(2) (PGE(2)) produced were measured using ELISAs. Results: Production of IL-8 by Ca9-22 cells was significantly increased when the cells were treated with OxLDL, but not with native LDL or acetylated LDL. Production of PGE(2) by Ca9-22 cells was enhanced by co-incubation with OxLDL and interleukin-1beta (IL-1beta). Scavenger receptor inhibitors, fucoidan and dextran sulfate, inhibited the OxLDL-induced IL-8 and PGE(2) production in the presence of IL-1beta. The p(38) MAPK inhibitors SB203580 and SB202190 and the ERK inhibitor PD98059 inhibited the OxLDL-induced IL-8 production. Among oxidized lipids and chemically modified LDL, 7-ketocholesterol enhanced IL-8 production. Conclusion: This is the first report to show that OxLDL enhances IL-8 production in epithelial cells.

60: Journal of neuroimmunology, 2010 Apr 16, 25(5)
IL8 and CXCL13 are potent chemokines for the recruitment of human neural precursor cells across brain endothelial cells.

[Abstract]It has been recently shown that systemically injected neural precursor cells (NPCs) could cross brain endothelium and favor functional recovery in animal models of multiple sclerosis (MS). Here we show that human NPCs express receptors of the chemokines IL8 and CXCL13 (CXCR1 and CXCR5, respectively) and migrate across brain endothelial cells in vitro, in response to these chemokines. Considering that these chemokines have been found overexpressed in MS in active, but not inactive areas of demyelination, our data suggest that systemically injected human NPCs may be considered for targeting active areas of demyelination in therapeutic approaches of MS.

61: Molecular nutrition & food research, 2010 Apr 15, 44(4)
Carbonyl compounds methylglyoxal and glyoxal affect interleukin-8 secretion in intestinal cells by superoxide anion generation and activation of MAPK p38.

[Abstract]The carbonyl compounds methylglyoxal (MG) and glyoxal (GL) are reactive intermediates of glucose degradation pathways and capable of inducing cellular damage. Although immune-stimulating activity has been investigated in endothelial cells, little is known about the signaling pathways of cytokine induction of these compounds in the intestine. Hence, we investigated the impact of mitogen-activated protein kinases (MAPK) and nuclear factor kappa B (NF-kappaB) on IL-8 production by human intestinal cells (Caco-2 and HT-29) after stimulation by MG and GL. Both compounds induced a dose-dependent enhancement of IL-8 secretion in human intestinal cells. MAPK p38 and extracellular signal-regulated kinase (ERK) were phosphorylated in these cells after having been stimulated by MG and GL. Furthermore, inhibitors of MAPK p38 (SB 203580 and 239063), ERK1/2 (PD 98059) and NF-kappaB activation (SM-7368 and SC-514) reduced IL-8 secretion. The most important mechanism by which MG and GL induced IL-8 secretion was the generation of superoxide anions which was confirmed by the inhibition of the cytosolic NADPH oxidase with diphenyl iodonium (DPI) or by application of superoxide dismutase (SOD). Our data suggest that multiple pathways were simultaneously activated; however, superoxide dependent MAPK p38 activation seems to be the most dominant pathway for IL-8 secretion in intestinal cells.

62: Critical care medicine, 2010 Apr 8, 34(5)
Plasma interleukin-8 is not an effective risk stratification tool for adults with vasopressor-dependent septic shock.

[Abstract]OBJECTIVE:: Plasma interleukin-8 levels of <220 pg/mL have an excellent negative predictive value (94% to 95%) for death at 28 days in children with septic shock and thus may be useful for risk stratification in clinical trial enrollment in this population. Whether plasma interleukin-8 would have similar use in adults with septic shock is unknown. DESIGN, SETTING, AND PATIENTS:: Analysis of plasma interleukin-8 levels in 192 adults with vasopressor-dependent septic shock enrolled in clinical trials of acute lung injury conducted by the Acute Respiratory Distress Syndrome Network. INTERVENTIONS:: XXXXX. MEASUREMENTS AND MAIN RESULTS:: Plasma interleukin-8 levels >/=220 pg/mL were significantly associated with death at 28 days in this cohort (odds ratio, 2.92; 95% confidence interval, 1.42 to 5.99; p = .001). However, in contrast to the findings in pediatric septic shock, a plasma interleukin-8 cutoff <220 pg/mL had a negative predictive value for death of only 74% (95% confidence interval, 66% to 81%) in adults with septic shock. Receiver operating characteristic analysis found an area under the curve of 0.59 for plasma interleukin-8, indicating that plasma interleukin-8 is a poor predictor of mortality in this group. In adults aged <40 yrs, a plasma interleukin-8 cutoff <220 pg/mL had a negative predictive value of 92%. CONCLUSIONS:: In contrast to similar pediatric patients, plasma interleukin-8levels are not an effective risk stratification tool in older adults with septic shock. Future studies of biomarkers for risk stratification in critically ill subjects will need to be replicated in multiple different populations before being applied in screening for clinical trials.

63: Journal of the European Academy of Dermatology and Venereology : JEADV, 2010 Apr 8, 34(5)
Influence of high dose tumescent local anaesthesia with prilocaine on systemic interleukin (IL)-6, IL-8 and tumour necrosis factor-alpha.

[Abstract]Abstract Background and objective Tumescent local anaesthesia (TLA) with high prilocaine doses leads to formation of methemoglobin (MHb) which is known to be a potent activator of pro-inflammatory endothelial cell response in vitro. As TLA is widely used for large dermatological resections, the aim of this study was to investigate the effects of high prilocaine doses on the systemic inflammatory response in vivo and its clinical relevance. Methods This prospective study examines the influence of MHb on serum interleukin (IL)-6, IL-8 and tumour necrosis tumour necrosis (TNF)-alpha levels up to 72 h after application of TLA with prilocaine in doses higher than 600 mg. Results A total of 30 patients received prilocaine in a median dose of 1500 mg (range: 880-4160 mg) for large resections. Peak prilocaine serum concentration was reached 4 h (0.72 +/- 0.07 mug/mL), the maximum concentration of MHb (7.43 +/- 0.87%) and IL-6 (28.4 +/- 4.1 U/L) 12 h after TLA application. TNF-alpha and IL-8 release were not found significantly increased. Three patients developed MHb concentrations >15%. Conclusions This clinical study shows for the first time that a high prilocaine serum concentration leads in vivo to elevated systemic levels of IL-6 but not of IL-8 and TNF-alpha because of initial high MHb levels. Because of possible and unpredictable high MHb concentrations, TLA should only be performed with prilocaine in doses of 2.5 mg/kg. In general, new solutions of TLA are necessary to achieve adequate anaesthesia for large dermatological resections to decrease the risk of methemoglobinaemia.

64: Blood, 2010 Jun 3, 115(22)
Toll-like receptor-induced reactivity and strongly potentiated IL-8 production in granulocytes mobilized for transfusion purposes.

[Abstract]Transfusion of granulocytes from granulocyte-colony stimulating factor (G-CSF)/dexamethasone (dexa)-treated donors can be beneficial for neutropenic recipients that are refractory to antimicrobial therapy. G-CSF/dexa treatment not only increases the number of circulating neutrophils but also affects their gene expression. Because of the intended transfusion of these granulocytes into patients who are severely ill, it is of importance to establish to what extent mobilization affects the cellular behavior of neutrophils. Here, we studied the effects of mobilization on Toll-like receptor (TLR)-mediated responses. Mobilized granulocytes displayed increased gene and protein expression of TLR2, TLR4, TLR5, and TLR8. Although mobilized granulocytes displayed normal priming of nicotinamide adenine dinucleotide phosphate oxidase activity and a slight increase in adhesion in response to TLR stimulation, these cells produced massive amounts of interleukin-8 (IL-8), in particular to TLR2 and TLR8 stimulation. The increase in IL-8 release occurred despite reduced IL-8 mRNA levels in the donor granulocytes after in vivo G-CSF/dexa treatment, indicating that the enhanced TLR-induced IL-8 production was largely determined by posttranscriptional regulation. In summary, granulocytes mobilized for transfusion purposes show enhanced TLR responsiveness in cytokine production, which is anticipated to be beneficial for the function of these cells on transfusion into patients.

65: Journal of periodontal research, 2010 Aug 1, 45(4)
Significance of elevated gingival crevicular fluid tumor necrosis factor-alpha and interleukin-8 levels in chronic hemodialysis patients with periodontal disease.

[Abstract]Da? A, Firat ET, Kadiro?lu AK, Kale E, Yilmaz ME. Significance of elevated gingival crevicular fluid tumor necrosis factor-alpha and interleukin-8 levels in chronic hemodialysis patients with periodontal disease. J Periodont Res 2010; 45: 445-450. (c) 2010 John Wiley & Sons A/S Background and Objective: The prevalence of chronic renal disease in industrialized countries is increasing, and chronic renal disease and periodontitis can have significant, reciprocal effects. The aim of this study was to evaluate the associations between specific clinical parameters and the levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) in the gingival crevicular fluid of hemodialysis (HD) patients with periodontal disease. Material and Methods: Forty-three HD patients and 43 systemically healthy subjects were enrolled in this study. Plaque index (PI), gingival index (GI) and pocket depth were used to determine periodontal status. Venous blood samples were obtained from each patient in the morning before the dialysis session and analyzed to determine the levels of inflammatory, biochemical and hematological parameters. Gingival crevicular fluid was collected from all subjects, and the levels of TNF-alpha and IL-8 were determined in the gingival crevicular fluid samples. Results: The following results were obtained from HD patients and controls: TNF-alpha (pg/mL), 31.40 +/- 1.46 and 3.06 +/- 0.15 (p < 0.001); IL-8 (pg/mL), 90.98 +/- 94.03 and 35.05 +/- 16.86 (p < 0.001); PI, 1.69 +/- 1.02 and 0.04 +/- 0.02 (p < 0.001); GI, 0.82 +/- 0.06 and 0.04 +/- 0.02 (p < 0.001); and pocket depth, 2.23 +/- 0.63 and 1.51 +/- 0.05 (p < 0.001), respectively. In addition, there were positive correlations between TNF-alpha and PI (r = 0.642, p < 0.001), between TNF-alpha and GI (r = 0.565, p < 0.001), between TNF-alpha and pocket depth (r = 0.522, p < 0.001), between IL-8 and PI (r = 0.402, p = 0.002), between IL-8 and GI (r = 0.396, p = 0.002), and between IL-8 and pocket depth (r = 0.326, p = 0.012). There were negative correlations between albumin and PI (r = -0.491, p < 0.001), albumin and GI (r = -0.406, p < 0.001), albumin and pocket depth (r = -0.464, p < 0.001) and albumin and CRP (r = -0.467, p = 0.002) and between the gingival crevicular fluid levels of TNF-alpha and IL-8, TNF-alpha and hemoglobin (r = -0.745, p < 0.001; r = -0.285, p < 0.05) (respectively). Conclusion: The levels of TNF-alpha and IL-8 in gingival crevicular fluid were significantly higher in HD patients than in controls. There were strong, positive correlations between clinical periodontal parameters and the levels of inflammatory cytokines in gingival crevicular fluid from the HD patients.

66: Journal of periodontal research, 2010 Aug 1, 45(4)
Interleukin-1 and interleukin-8 in nicotine- and lipopolysaccharide-exposed gingival keratinocyte cultures.

[Abstract]Johnson GK, Guthmiller JM, Joly S, Organ CC, Dawson DV. Interleukin-1 and interleukin-8 in nicotine- and lipopolysaccharide-exposed gingival keratinocyte cultures. J Periodont Res 2010; 45: 583-588. (c) 2010 John Wiley & Sons A/S Background and Objective: Tobacco use is associated with increased periodontal destruction in both cigarette smokers and smokeless tobacco users. Gingival keratinocytes are the first cells in contact with microbial and tobacco components and play a key role in the innate immune response to these agents. The objective of this study was to evaluate the effect of nicotine and bacterial lipopolysaccharide (LPS) alone and in combination on gingival keratinocyte production of interleukin-1alpha (IL-1alpha) and interleukin-8 (IL-8). Material and Methods: Gingival keratinocyte cultures were established from 10 healthy, non-tobacco-using subjects. The cells were stimulated for 24 h with 1 mum or 1 mm nicotine and/or 10 mug/mL Escherichia coli or Porphyromonas gingivalis LPS. Interleukin-1alpha and IL-8 proteins were quantified using ELISAs. Results: Compared with untreated cultures, 1 mm nicotine stimulated production of IL-1alpha (p < 0.001); E. coli and P. gingivalis LPS increased IL-8 production (p = 0.0014 and p = 0.0232, respectively). A combination of nicotine and LPS produced the highest cytokine quantities. Amounts of IL-1alpha and IL-8 following 1 mm nicotine and LPS exposure were significantly greater than in untreated cultures (p < 0.001). Interleukin-8 was also responsive to 0.1 mum nicotine combined with E. coli or P. gingivalis LPS compared with control cultures (p < 0.0001 and p = 0.0029, respectively). Both cytokines tended to be elevated following the combined treatment relative to nicotine or LPS treatment alone. Conclusion: These results demonstrate that nicotine and LPS differentially regulate IL-1 and IL-8 production by gingival keratinocytes. Combined treatment tended to elevate cytokine production further, which may have implications for the progression of periodontitis in tobacco users.

67: Biochimica et biophysica acta, 2010 Mar 18, 139(4)
Galactose specific adhesin of enteroaggregative E.coli induces IL-8 secretion via activation of MAPK and STAT-3 in INT-407 cells.

[Abstract]BACKGROUND: Enteroaggregative Escherichia coli (EAEC) is one of the most common bacterial pathogen associated with the etiology of persistent diarrhea. A characteristic feature of EAEC-pathogenesis is the induction of profound inflammatory response in the intestinal epithelium. The present study was designed to investigate the underlying mechanism of inflammatory responses induced by a novel galactose specific adhesin of T7 strain of EAEC (EAEC-T7) in human intestinal epithelial cell line (INT-407). METHODS: INT-407 cells were stimulated with the adhesin in absence and presence of anti-adhesin (IgG(AD))/ D-galactose / H7 / staurosporin (inhibitor of PKC) / PD098059 (inhibitor of MEK) / SB203580 (inhibitor of p38-MAPkinase) / AG490 (inhibitor of JAK (-2,-3)/STAT-3 pathway). The expression of activated Raf-1, MEK-1, ERK1/2, JNK, p38-MAPK and STAT-3 was analyzed by Western immunoblot. Release of Interleukin-8 (IL-8) was measured by ELISA. RESULTS: The adhesin was found to induce activation of Raf-1, MEK-1, ERK1/2, p38-MAPK and STAT-3, which was reduced in presence of IgG(AD) / D-galactose. The activation of Raf-1 was found to be attenuated in presence of H7/staurosporin. The expression of phosphorylated STAT-3 was downregulated in presence of AG490 and PD098059. Further, the adhesin induced IL-8 secretion was reduced in presence of the inhibitors of MEK (PD098059), p38-MAPK (SB203580) and JAK (-2,-3)/STAT-3 pathway (AG490). CONCLUSIONS: We propose that STAT-3 activation is quintessential for the galactose specific adhesin induced IL-8 secretion by INT-407 cells and must occur in concert with the activation of ERK1/2. GENERAL SIGNIFICANCE: Our contribution regarding the galactose specific adhesin mediated signaling leads to an improved understanding of the EAEC- pathogenesis and may provide novel therapeutic approaches to combat EAEC- infection.

68: Chemico-biological interactions, 2010 May 14, 185(3)
Mycophenolic acid inhibits the phosphorylation of NF-kappaB and JNKs and causes a decrease in IL-8 release in H2O2-treated human renal proximal tubular cells.

[Abstract]Ischaemia-reperfusion injury is a common occurrence in renal transplantation and may affect the long-term survival of the allograft. Oxidative stress may play a crucial role in this, with reactive oxygen species formed during reperfusion causing direct cellular damage as well as activating pro-inflammatory pathways. A human proximal tubule cell line (HK-2) was subjected to hydrogen peroxide (H(2)O(2)) stress that resulted in phosphorylation of c-jun N-terminal kinases (JNKs) and the transcription factor NF-kappaB at Ser276, both of which have been associated with inflammation. Interleukin (IL)-8 production also increased upon H(2)O(2) stimulation. Pre-incubation of the cells with mycophenolic acid (MPA) resulted in reduced phosphorylation of both JNKs and NF-kappaB, and reduced IL-8 release in H(2)O(2)-stimulated HK-2 cells. MPA also reduced the H(2)O(2)-induced phosphorylation of p38 MAP (mitogen-activated protein) kinase, the extracellular-signal regulated kinase 1/2 (ERK1/2), Akt kinase and the transcription factor CREB (cyclic AMP response element binding protein). In rat kidneys subjected to ischaemia-reperfusion, an increase in both pJNK1/2 and pNF-kappaB was observed, which was reduced in kidneys obtained from mycophenolate mofetil (MMF)-treated rats. These results suggest that MPA may inhibit pro-inflammatory responses in the kidney by inhibiting activation of pro-inflammatory molecules in both the kidney and human renal proximal tubular cells subjected to oxidative stress.

69: Molecular biology reports, 2010 Mar 19, 337(1-2)
Interleukin-8-251A/T polymorphism and Helicobacter pylori infection influence risk for the development of gastric cardiac adenocarcinoma in a high-incidence area of China.

[Abstract]Polymorphisms in cytokine genes may contribute to increased susceptibility to different cancers. The aim of this paper is to investigate the association of IL-8-251A/T polymorphism and Helicobacter pylori (H. pylori) infection with the risk of developing gastric cardiac adenocarcinoma (GCA) in the south of Taihang Mountain, a high-incidence area of esophageal cancer in China. The IL-8-251 A/T polymorphism was genotyped in 519 cases of GCA and 504 healthy controls. The H. pylori infection in GCA patients and controls was detected by rapid urease test (RUT), histopathology or (14)C-urea breath test ((14)C-UBT). The results showed that family history of upper gastrointestinal cancer (UGIC) and H. pylori infection significantly increased the risk of developing GCA. The overall genotype and allelotype distributions of IL-8 promoter SNPs in GCA patients were significantly different from those in healthy controls. Compared with TT genotype, AA genotype significantly elevated the risk of developing GCA. The stratification analysis revealed that, compared with the TT genotype, the AA genotype significantly elevated the risk of developing GCA in both positive family history of UGIC and H. pylori infection subgroups. This study provides evidence to support a relationship of increased susceptibility to GCA in individuals of the south Taihang Mountain region with IL-8 251 AA genotype, especially for those individuals who have family history of UGIC or H. pylori infection.

70: Journal of inflammation (London, England), 2010 Mar 18, 7(1)
Cytosolic group IVa phospholipase A2 mediates IL-8/CXCL8-induced transmigration of human polymorphonuclear leukocytes in vitro.

[Abstract]ABSTRACT: BACKGROUND: Cytosolic gIVaPLA2 is a critical enzyme in the generation of arachidonate metabolites and in induction of beta 2-integrin adhesion in granulocytes. We hypothesized that gIVaPLA2 activation also is an essential downstream step for post adhesive migration of PMN in vitro. METHODS: Migration of PMNs caused by IL-8/CXCL8 was assessed using a transwell migration chamber. PMNs were pretreated with two structurally unrelated inhibitors of gIVaPLA2, arachidonyl trifluoromethylketone (TFMK) or pyrrophenone, prior to IL-8/CXCL8 exposure. The fraction of migrated PMNs present in the lower chamber was measured as total myeloperoxidase content. GIVaPLA2 enzyme activity was analyzed using [14C-PAPC] as specific substrate. F-actin polymerization and cell structure were examined after rhodamine-phalloidin staining. RESULTS: IL-8/CXCL8-induced migration of PMNs was elicited in concentration- and time-dependent manner. Time-related phosphorylation and translocation of cytosolic gIVaPLA2 to the nucleus was observed for PMNs stimulated with IL-8/CXCL8 in concentration sufficient to cause upstream phosphorylation of MAPKs (ERK-1/2 and p38) and Akt/PKB. Inhibition of gIVaPLA2 corresponded to the magnitude of blockade of PMN migration. Neither AA nor LTB4 secretion was elicited following IL-8/CXCL8 activation. In unstimulated PMNs, F-actin was located diffusely in the cytosol; however, a clear polarized morphology with F-actin-rich ruffles around the edges of the cell was observed after activation with IL-8/CXCL8. Inhibition of gIVaPLA2 blocked change in cell shape and migration caused by IL-8/CXCL8 but did not cause F-actin polymerization or translocation of cytosolic F-actin to inner leaflet of the PMN membrane. CONCLUSION: We demonstrate that IL-8/CXCL8 causes a) phosphorylation and translocation of cytosolic gIVaPLA2 to the nucleus, b) change in cell shape, c) polymerization of F-actin, and d) migration of PMN in vitro. Inhibition of gIVaPLA2 blocks the deformability and subsequent migration of PMNs caused by IL-8/CXCL8. Our data suggest that activation of gIVaPLA2 is an essential step in PMN migration in vitro.

71: Journal of enzyme inhibition and medicinal chemistry, 2010 Mar 17, 7(1)
Novel depsides as potential anti-inflammatory agents with potent inhibitory activity against Escherichia coli-induced interleukin-8 production.

[Abstract]Sixteen novel depsides were synthesized for the first time. Their chemical structures were clearly determined by (1)H NMR, ESI mass spectra, and elemental analyses. All the compounds were assayed for antibacterial activities against three Gram-positive bacterial strains (Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 6538, and Streptococcus faecalis ATCC 9790) and three Gram-negative bacterial strains (Escherichia coli ATCC 35218, Pseudomonas aeruginosa ATCC 13525, and Enterobacter cloacae ATCC 13047) by the MTT method. Compound 2-(2-methoxy-2-oxoethyl)phenyl 5-bromonicotinate (5) exhibited significant antibacterial activities against E. coli ATCC 35218 with an MIC of 0.78 mug/mL, which was superior to the positive control kanamycin B. In addition, compound 5 showed potent inhibitory activity against E. coli-induced interleukin-8 production.

72: The Journal of investigative dermatology, 2010 Apr, 130(4)
EGFR (Trans)activation Mediates IL-8 and Distinct Human Antimicrobial Peptide and Protein Production following Skin Injury.

[Abstract]Antimicrobial peptides and proteins (AMPs) are tools of the innate immune system employed at injury sites to protect the host from invading microbes and to promote wound repair. In this issue, Roup�� et al. characterize epidermal innate immune responses induced by skin injury and the involvement of EGFR for distinct AMPs and IL-8 induction.

73: Actas dermo-sifiliograficas, 2010 Mar, 101(2)
Production of interleukin-8 by circulating CLA+ T cells with skin tropism in patients with psoriasis and in healthy controls.

[Abstract]BACKGROUND: Psoriasis is an immune-mediated disease typically associated with cutaneous neutrophilic infiltration and Munro microabscesses. Interleukin (IL)-8 is one of the main neutrophil-attracting chemokines. Although keratinocytes have traditionally been considered to be the principal source of IL-8 in psoriasis, we present data that suggest that cutaneous lymphocyte associated antigen (CLA) + T lymphocytes synthesize this cytokine. MATERIAL AND METHODS: Six patients with psoriasis and 6 healthy controls were studied. Immunomagnetic separation was used to isolate CLA+ and CLA- T lymphocytes and IL-8 and interferon (IFN)-gamma production was quantified for each cell subpopulation using enzyme-linked immunosorbent assay. Finally, gene expression of IL-8 was analyzed by reverse transcriptase-polymerase chain reaction. RESULTS: CLA+ and CLA- T lymphocytes from patients with psoriasis and from controls showed a significantly increased production of IFN-gamma when activated, whereas only activated CLA+ T lymphocytes (from patients and controls) synthesized IL-8. The higher level of expression of IL-8 and IFN-gamma by CLA+T lymphocytes in comparison to CLA- cells was confirmed. DISCUSSION: Previous studies have confirmed IL-8 production by T lymphocytes in inflammatory skin diseases with neutrophil-rich infiltrates, such as acute generalized exanthematous pustulosis, Beh?et disease, and pustular psoriasis. We have confirmed the role of the subset of T lymphocytes with skin tropism (CLA+) in IL-8 production in nonpustular psoriasis.

74: International journal of oral and maxillofacial surgery, 2010 Mar 4, 25(4)
Proinflammatory cytokines (IL-1beta and TNF-alpha) and chemokines (IL-8 and MIP-1alpha) as markers of peri-implant tissue condition.

[Abstract]Analysis of peri-implant crevicular fluid (PICF) offers a non-invasive means of studying the host response in peri-implant disease and may provide an early indication of patients at risk for active disease. This study examined the PICF levels of interleukin-1beta (IL-1beta), tumour necrosis factor alpha (TNF-alpha), interleukin-8 (IL-8) and macrophage inflammatory protein-1alpha (MIP-1alpha) in patients with non-manifesting inflammation, early and late stages of mucositis. The study group comprised 90 adult healthy volunteers with endosseal titanium implants inserted. Samples were taken from peri-implant sulcus using a filter paper technique. Implant tissues were categorized clinically as healthy, early mucositis or advanced mucositis. Clinical manifestations were determined by: gingival index and bleeding on probing, plaque index and radiographic analyses. Cytokine concentrations were assesed using commercially available enzyme-linked immunosorbent assay kits. Patients from the control group (healthy patients) have significantly lower concentrations of IL-1beta, TNF-alpha, IL-8 and MIP-1alpha in PICF compared with both groups with mucositis. Positive correlation was noted in the control group between IL-1beta and TNF-alpha and between MIP-1alpha and IL-8 in the group with early mucositis. The results suggest that cytokines could be prognostic markers of implant failure.

75: Cytokine, 2010 Mar 3, 25(4)
The -251A>T polymorphism of interleukin-8 is associated with longer mechanical ventilation and hospital staying after coronary surgery.

[Abstract]Background: Cardiac surgery is associated with inflammatory responses that are known to affect its outcome. The present study was designed to define whether post-operative release of interleukin (IL)-6, 8 and tumor necrosis factor-alpha (TNF-alpha) is related to the presence of a certain allele in functional polymorphism and its relationship to clinical outcome after off-pump coronary artery bypass (OPCAB). Methods: One hundred and forty-five patients undergoing first time elective OPCAB were genotyped for IL-6(-174G>C), IL-8(-251A>T) and TNF-alpha(-308G>A) polymorphisms using polymerase chain reaction (PCR) and gene sequencing. Cytokine levels were measured in plasma samples taken before the operation and 4, 24 and 72h postoperatively by suspension array system. Results: Levels of IL-6 and IL-8 increased significantly after OPCAB. Patients with IL-6-174GG and IL-8-251AA genotypes had higher post-operative circulating levels of IL-6 and IL-8, respectively. Logistic regression showed that IL-8-251AA genotype was an independent risk factor of ventilation time more than 1day (OR=11.80, 95% CI: 1.87-74.48) and hospital staying more than 14days (OR=38.00, 95% CI: 4.15-347.87) after surgery. Conclusions: OPCAB results in post-operative inflammatory responses. Genetic backgrounds alter the extent of inflammatory response and might relate to clinical outcome of OPCAB.

76: Blood cells, molecules & diseases, 2010 Mar 1, 79(5)
Effect of interleukin-8 and RANTES on the Gardos channel activity in sickle human red blood cells: Role of the Duffy antigen receptor for chemokines.

[Abstract]We investigated the effects of the chemokines IL-8 and RANTES on the activity of the Gardos channel (GC) of sickle red blood cells (SSRBCs). SSRBCs expressing the Duffy antigen receptor for chemokines (DARC) incubated under oxygenated conditions exhibit GC activation. The deoxygenation-stimulated K(+) loss via the GC is activated by the chemokines in the Duffy-positive SSRBCs. The percentage of cells with high density is 17 times higher in the Duffy-positive group. These findings are consistent with a greater susceptibility of Duffy-positive SSRBCs to inflammatory chemokines leading to GC activation and cellular dehydration and suggest a coupling, promoted by the sickling process, between DARC and the GC.

77: International journal of molecular medicine, 2010 Apr, 25(4)
Involvement of capsular polysaccharide via a TLR2/NF-kappaB pathway in Vibrio vulnificus-induced IL-8 secretion of human intestinal epithelial cells.

[Abstract]In a previous study, we reported that a wbpP gene mutation in Vibrio vulnificus was significantly impaired in its ability to synthesize surface capsular polysaccharide (CPS). In this study, we evaluated the functions of the V. vulnificus capsular polysaccharide on interleukin (IL)-8 production, as well as its underlying mechanisms in human intestinal epithelial cells. The CPS-defective wbpP mutant induced significantly lower levels of IL-8 production, IL-8 gene promoter activation and NF-kappaB activity in INT-407 cells than was noted with the wild-type or wbpP-complemented V. vulnificus. The expression levels of Toll-like receptor (TLR)2 mRNA and protein were also found to be lower in INT-407 cells infected with the CPS-defective wbpP mutant than in those cells infected with the wild-type or the wbpP-complemented strains. Additionally, the treatment of INT-407 cells with anti-TLR2 antibody proved to significantly block IL-8 production and NF-kappaB minimal promoter activity induced by the wild-type or the wbpP-complemented strains. Furthermore, purified V. vulnificus CPS was found to significantly induce IL-8 production and NF-kappaB activation, both of which were inhibited upon the addition of the anti-TLR2 antibody. Taken together, these results demonstrate that V. vulnificus capsular polysaccharide is involved in the induction of IL-8 production of human intestinal epithelial cells via a TLR2/NF-kappaB-dependent pathway.

78: Journal of immunology (Baltimore, Md. : 1950), 2010 Mar 1, 16(3)
Proinflammatory Cytokine IL-1{beta} Stimulates IL-8 Synthesis in Mast Cells via a Leukotriene B4 Receptor 2-Linked Pathway, Contributing to Angiogenesis.

[Abstract]Recent studies have suggested that mast cells have critical roles in angiogenesis. However, the detailed mechanism by which mast cells contribute to angiogenesis is not yet clearly understood, especially in response to proinflammatory cytokines. In this study, we showed that the proinflammatory cytokine IL-1beta induces the synthesis of IL-8, a potent angiogenic factor, in human mast cells via the leukotriene B(4) receptor (BLT)2. We also characterized the BLT2 downstream signaling pathway and determined that BLT2-mediated IL-8 synthesis involves the upregulation of Nox1, a member of the NADPH oxidase family, Nox1-dependent reactive oxygen species generation and the subsequent activation of the redox-sensitive transcription factor NF-kappaB. For instance, knockdown of BLT2 and Nox1 with specific small interfering RNA, treatment with a specific BLT2 antagonist, LY255283, or treatment with a potential Nox inhibitor, diphenylene iodonium, suppressed IL-1beta-induced IL-8 synthesis. We found that the conditioned media collected from IL-1beta-treated human mast cell line HMC-1 had significantly enhanced angiogenic activity that could be dramatically attenuated by either small interfering RNA knockdown of BLT2 or treatment with neutralizing Ab to IL-8. Finally, the experiments were repeated using human primary cord blood-derived mast cells, and the results were clearly reproduced. Taken together, our results suggest that BLT2-Nox1-reactive oxygen species-dependent pathway plays a role in promoting the secretion of IL-8 from human mast cells in response to the proinflammatory cytokine IL-1beta, thus contributing to angiogenesis.

79: Environmental health perspectives, 2010 Mar 1, 34(3)
Phosphorylation of p65 Is Required for Zinc Oxide Nanoparticle-induced Interleukin 8 Expression in Human Bronchial Epithelial Cells.

[Abstract]BACKGROUND: Exposure to zinc oxide (ZnO) in environmental and occupational settings causes acute pulmonary responses through the induction of pro-inflammatory mediators such as interleukin-8 (IL-8). OBJECTIVE: We investigated the effect of ZnO nanoparticles on IL-8 expression and the underlying mechanisms in human bronchial epithelial cells. METHODS: IL-8 mRNA and protein expression in primary human bronchial epithelial cells and the human bronchial epithelial cell line, BEAS-2B, was determined with RT-PCR and ELISA, respectively. Transcriptional activity of IL-8 promoter and NFkappaB in ZnO-treated BEAS-2B cells was measured using transient gene transfection of the luciferase reporter construct with or without p65 constructs. IkappaBalpha phosphorylation and degradation, and p65 phosphorylation were detected using immunoblotting. Binding of p65 to the IL-8 promoter was examined using chromatin immunoprecipitation assay. RESULTS: ZnO exposure (2-8 mug/ml) increased IL-8 mRNA and protein expression. Transcription inhibition with actinomycin D blocked ZnO-induced IL-8 expression, which was consistent with the observation that ZnO exposure increased IL-8 promoter reporter activity. Further study demonstrated that the kappaB-binding site in the IL-8 promoter was required for ZnO-induced IL-8 transcriptional activation. ZnO stimulation modestly elevated IkappaBalpha phosphorylation and degradation. Moreover, ZnO exposure also increased the binding of p65 to the IL-8 promoter and p65 phosphorylation at serines 276 and 536. Overexpression of p65 constructs mutated at serines 276 or 536 significantly reduced ZnO-induced increase in IL-8 promoter reporter activity. . CONCLUSION: p65 phosphorylation and IkappaBalpha phosphorylation and degradation are the primary mechanisms involved in ZnO nanoparticle-induced IL-8 expression in human bronchial epithelial cells.

80: Biochimica et biophysica acta, 2010 Feb 23, 23(3)
Selenium supplementation attenuates procollagen-1 and interleukin-8 production in fat-loaded human C3A hepatoblastoma cells treated with TGFbeta1.

[Abstract]BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is associated with obesity, insulin resistance and hepatic steatosis. Non-alcoholic steatohepatitis (NASH) is a serious consequence of NAFLD where chronic tissue damage and inflammation result in fibrosis which may progress to cirrhosis. Transforming growth factor beta1 (TGFbeta1), proinflammatory cytokines and oxidative stress are thought to play crucial roles in the pathogenesis of these conditions. The contributions of individual liver cell types to fibrogenesis remain controversial and the influence of selenium status has not been investigated. METHODS: In this study we have used a cell culture model of fat-loading using oleate-treated human hepatoblastoma (C3A) cells to investigate how fat-loading and selenium status might influence the production of collagen in response to TGFbeta1. The secretion of inflammatory cytokines was also investigated, together with the epithelial character of the treated cells. RESULTS: We found that in response to treatment with TGFbeta1, C3A cells produced mRNA encoding the pro-alphaI chain of procollagen I, secreted procollagen I peptide, up-regulated production of the proinflammatory cytokine interleukin-8 (IL-8) and the mesenchymal marker vimentin, and down-regulated albumin production. Most of these responses were considerably enhanced when cells were fat-loaded with oleate and were attenuated by selenium addition at a dose that optimised the expression of thioredoxin reductase and glutathione peroxidase. CONCLUSIONS: Our data establish that both fat-loading and suboptimal selenium status enhance collagen and IL-8 production by C3A hepatocytes in response to TGFbeta1, possibly as part of an epithelial to mesenchymal transition. GENERAL SIGNIFICANCE: These findings suggest that the hepatocyte may be an important contributor to the pathogenesis of fibrosis associated with NAFLD.

81: Experimental eye research, 2010 Feb 23, 23(3)
Induction of interleukin-8 Gene Expression and Protein Secretion by C-reactive protein in ARPE-19 Cells.

[Abstract]C-reactive protein (CRP) is an acute phase reactant and its level rises rapidly during inflammation. Recent studies have suggested the potential involvement of CRP in the pathogenesis of age-related macular degeneration (AMD). To delineate the functional roles of CRP in inflammatory response by the ocular posterior segments, the effects of CRP on ARPE-19, an immortalized human retinal pigment epithelia (hRPE) cell line, were investigated in the present study. Treatment of ARPE-19 cells with CRP resulted in enhanced NF-kB nuclear translocation and dose-dependent transient induction of IL-8 mRNA synthesis and protein secretion. Stimulated expression of VEGF, but not MCP-1 by CRP was also observed. The induced IL-8 expression was transient and peaked at 12 h post stimulation. In the presence of inhibitors for NF-kB, p38, MEK and JNK, the CRP-induced IL-8 production was abolished by 99.5+/-2.3, 97.8+/-2.1, 55.3+/-2.5 and 37.3+/-1.3%, respectively. Neutralization of Fc gamma receptors by anti-CD32 and CD64 antibodies produced 39.9+/-1.6 and 59.5+/-2.6% reduction, respectively, of CRP-stimulated IL-8 secretion, whereas that by anti-CD16 antibody had no effect. This study suggested that the pro-inflammatory effects of CRP in ARPE-19 cells may contribute to the inflammatory retinal diseases by induction of pro-inflammatory cytokines such as IL-8. This induction is mediated by NF-kB and multiple MAPK pathways through Fc gamma receptors.

82: American journal of physiology. Cell physiology, 2010 Feb 24, 23(3)
ROCK2 Associates with Lectin-like Oxidized-LDL Receptor-1 (LOX-1) and Mediates OxLDL-induced IL8 Production.

[Abstract]Oxidatively modified low density lipoprotein (OxLDL) is a contributing factor of endothelial dysfunction, an early cellular event during atherogenesis. In endothelial cells, OxLDL has been shown to stimulate proinflammatory responses, increase lipid accumulation, and induce the expression of adhesion and extracellular matrix degrading molecules. The primary receptor for OxLDL on endothelial cells has been identified as a member of the scavenger receptor family called Lectin-like Oxidized-LDL Receptor-1 (LOX-1). A number of studies on LOX-1 have implicated its role in multiple cardiovascular diseases including atherosclerosis. In order to better understand the molecular mechanisms underlying the role of LOX-1 in endothelial cells, we identified interacting proteins in an affinity-purified LOX-1 receptor complex from human aortic endothelial HAECT cells by mass spectrometry. Two molecules involved in Rho signaling pathway, ARHGEF1 and ROCK2, were identified and their associations with LOX-1 were confirmed in reciprocal immunoprecipitation studies. Particularly, ROCK2 was found to dynamically associate with LOX-1 in the presence of OxLDL. In addition, OxLDL treatment stimulated ROCK2 catalytic activity, and ROCK2 inhibition attenuated NF-kappaB activation and IL-8 production resulting from OxLDL activation of LOX-1. In summary, a functional proteomics approach has enabled us to identify novel LOX-1 interactors that potentially contribute to the cellular and signaling functions of LOX-1.

83: Molecular biology reports, 2010 Feb 21, 23(3)
Effects of 15-deoxy-(12,14)-prostaglandin J (2) on the production of IL-8 and the expression of Toll-like receptor 2 in human primary keratinocytes stimulated with lipopolysaccharide.

[Abstract]15-deoxy-(12,14)-prostaglandin J(2) (15d-PGJ(2)) is an anti-inflammatory prostaglandin that plays a role in promoting the resolution of inflammation. We investigated the effects of 15d-PGJ(2) on the production of IL-8 and on the expression of Toll-like receptors (TLRs) 2 in human primary keratinocytes stimulated with lipopolysaccharide (LPS). Cell proliferation was analyzed using the MTT assay, TLR2 and -4 mRNA expression was detected by RT-PCR, and IL-8 production and NF-kappaB p65 activities were determined by ELISA. LPS and 15d-PGJ(2) did not influence the proliferation rate at low concentrations (0.5 and 2.0 muM) in keratinocytes, and showed toxicity at high concentrations (5.0 muM). LPS, compared with control, induced the expression of TLR2 mRNA, increased IL-8 production, and enhanced NF-kappaB activity. 15d-PGJ(2) decreased TLR2 mRNA, increased IL-8 production, and suppressed NF-kappaB activity. Costimulation with LPS and 15d-PGJ(2), compared with LPS stimulation alone, decreased TLR2 mRNA (1.8-fold), increased IL-8 production (1.8-fold at 0.5 muM and 3.7-fold at 2.0 muM), and inhibited NF-kappaB activity (3.3-fold at 0.5 muM and 5.1-fold at 2.0 muM). TLR4 mRNA was not expressed in primary keratinocytes. These results suggest that 15d-PGJ(2) suppresses TLR2 expression and that it up-regulates the production of IL-8 by inhibiting the NF-kappaB signaling pathway in primary keratinocytes. Thus, 15d-PGJ(2) can have both anti- and pro-inflammatory effects, and 15d-PGJ(2)-mediated IL-8 up-regulation is related to the mitogen-activated protein kinase (MAPK) and NF-kappaB signaling pathways.

84: Toxicology letters, 2010 Feb 18, 23(3)
Involvement of p120 in LPS-induced NF-kappaB activation and IL-8 production in human bronchial epithelial cells.

[Abstract]P120-catenin (p120), a prototypic member of a subfamily of Armadillo repeat domain (Arm domain) proteins, not only participates in cell-cell adhesion, but also mediates inflammatory responses in the skin. In the present study, we demonstrated the effect of p120 on lipopolysaccharide (LPS)-induced inflammatory responses in human bronchial epithelial cells (BECs). We first confirmed that p120 expression was significantly reduced after LPS stimulation in BECs, the p65 subunit of nuclear factor-kappaB (NF-kappaB) nuclear translocation was promoted and NF-kappaB activity was rapidly induced. Moreover, the expression level of interleukin-8 (IL-8) increased after LPS treatment. Over-expression of p120 attenuated LPS-stimulated NF-kappaB reporter gene expression and IL-8 mRNA expression and protein synthesis. On the contrary, transfection with p120 small interfering RNA (siRNA) significantly elevated LPS-stimulated NF-kappaB transcriptional activity, p65 nuclear translocation and IL-8 expression. Collectively, theses results indicate an anti-inflammatory effect of p120 in BECs, through its modulation of NF-kappaB signaling.

85: The Journal of pediatrics, 2010 Feb 18, 23(3)
Elevated Serum IL-8 Levels in Ataxia Telangiectasia.

[Abstract]Serum interleukin (IL)-8 levels were measured in 50 patients with ataxia telangiectasia (A-T) and 22 without A-T. In a cross-sectional study, the geometric mean of IL-8 level was significantly higher in the patients with A-T (P <.0001). Elevated serum IL-8 levels in patients with A-T suggest that systemic inflammation may contribute to the disease phenotype.

86: Free radical research, 2010 Feb 19, 23(3)
High cerebrospinal fluid antioxidants and interleukin 8 are protective of hypoxic brain damage in newborns.

[Abstract]Abstract The objective was to explain the discrepancy in the development of hypoxic ischemic brain injury (HIE) in some asphyxiated newborns rather than others. Forty newborns were classified according to their cerebrospinal neuron-specific-enolase (CSF-NSE) levels on their 5th-day of life; group 1 with low-NSE (n = 25). The remaining 15 newborns had high-NSE and were further divided into a group with no HIE (n = 10, group 2) and another with HIE (n = 5, group 3). CSF-NSE, totalhydroperoxide (TH), biological-antioxidant-potentials (BAPs), 12 cytokines and Erythropoietin (EPO) were measured. The TH/BAP gave the oxidative-stress-index (OSI). The BAPs of serial dilutions of three types of EPO were tested. CSF-NSE and TH and mean OSIs were higher in group 3. IL-8 and mean BAPs were higher in group 2 than in group 1. EPO was less detected in group 3. Serial EPO dilutions correlated with their BAPs. Compensatory antioxidants and IL-8 elevation could be protective of perinatal asphyxic brain injury. Antioxidative effect of EPO could be neuroprotective.

87: Journal of acquired immune deficiency syndromes (1999), 2010 Feb 15, 316(4)
Resistance to Simian HIV Infection Is Associated With High Plasma Interleukin-8, RANTES and Eotaxin in a Macaque Model of Repeated Virus Challenges.

[Abstract]Animal models for research on susceptibility to HIV are currently not available. Here we explore whether a macaque model of repeated low-dose rectal or vaginal virus challenges could be employed. We tested the hypothesis that susceptibility to Simian HIV is not merely stochastic in this model but rather is associated with identifiable host factors. Forty macaques required a median of 3.5 SHIVSF162P3 challenges for infection. We studied the association of their susceptibility with 13 predisposing plasma cytokines/chemokines (RANTES, Eotaxin, monocyte chemoattractant protein (MCP)-1, IL-7, MIP-1beta, TNF-alpha, MIP-1alpha, granulocyte colony-stimulating factor, IL-8, interferon-gamma, IL-17, IL-1beta, IL-6). Higher plasma RANTES, IL-8, and Eotaxin were associated with lower susceptibility, that is, higher resistance to infection. In a group of macaques with low IL-8 and RANTES, a median 3 exposures were required to infect; whereas, when either IL-8 or RANTES were high, a median 12 exposures were required. Thus, susceptibility was associated with identifiable discrete host factors and was not stochastic. In addition, the macaque model identified key human resistance factors (RANTES, Eotaxin), but also revealed a novel association with resistance (IL-8). Future direct evaluation of these or other factors in the animal model may be beneficial for developing new immunomodulation strategies for HIV prevention.

88: The Journal of pharmacology and experimental therapeutics, 2010 Feb 17, 23(3)
Butrin, Isobutrin and Butein from Medicinal Plant Butea monosperma Selectively Inhibit NF-{kappa}B in Activated Human Mast Cells: Suppression of TNF-{alpha}, IL-6 and IL-8.

[Abstract]Activation of mast cells in rheumatoid synovial tissue has often been associated with tumor necrosis factor (TNF)-alpha, interleukin (IL)-6 and IL-8 production and disease pathogenesis by adjacent cell types. Butea monosperma (BM), is a well known medicinal plant in India and the tropics. The aim of this study was to examine whether a standardized extract of BM flower (BME) could inhibit inflammatory reactions in human mast cells using activated HMC-1 cells as a model. Four previously characterized polyphenols- butrin, isobutrin, isocoreopsin and butein- were isolated from BME by preparative TLC and their purity and molecular weights were determined by LC/MS analysis. Our results showed that butrin, isobutrin and butein significantly reduced the PMACI-induced inflammatory gene expression and production of TNF-alpha, IL-6 and IL-8 in HMC-1 cells by inhibiting the activation of NF-kappaB. In addition, isobutrin was most potent in suppressing the NF-kappaB p65 activation by inhibiting IkappaBalpha-degradation, whereas butrin and butein were relatively less effective. In-vitro kinase activity assay revealed that isobutrin was a potent inhibitor of IKK activity. Ours is the first report identifying the molecular basis of the reported anti-inflammatory effects of BME and its constituents butrin, isobutrin and butein. The novel pharmacological actions of these polyphenolic compounds indicate potential therapeutic value for the treatment of inflammatory and other diseases in which activated mast cells play a role.

89: International journal of colorectal disease, 2010 Feb 17, 23(3)
Differential expression of the chemokines GRO-2, GRO-3, and interleukin-8 in colon cancer and their impact on metastatic disease and survival.

[Abstract]BACKGROUND AND AIM: Chemotactic cytokines play a role in angiogenesis and attraction of immune cells. However, their contribution to tumor formation remains incompletely understood. In a previous transcriptome study, we identified a family of structurally related chemokines of the CXC-family to be specifically up-regulated in colorectal cancer. The aim of the present study was to investigate the regulation of their expression in colon cancer cells and to test the hypothesis that altered CXC-chemokine expression is related to critical clinical parameters, such as survival or metastasis formation. MATERIALS AND METHODS: Expression levels of interleukin-8 (CXCL-8) and growth-related oncogenes 2 and 3 (GRO-2/CXCL-2 and GRO-3/CXCL-3) were quantified using qRT-PCR in 97 patients with completely resected colon carcinoma and correlated with clinical parameters. Moreover, 16 samples of normal mucosa, nine samples of benign adenoma, and 11 samples of liver metastasis were analyzed. Next, the regulation of chemokine expression in response to various stimuli was tested in colon cancer cell lines (HT29, HCT116, CaCO2). RESULTS: Expression of GRO-2, GRO-3, and IL-8 was significantly increased in colon cancer as compared to normal colon tissue. Expression of GRO-2 and GRO-3 was already enhanced in premalignant adenomas, and GRO-3 was significantly down-regulated in liver metastasis as compared to the primary tumor. Importantly, expression of GRO-3 was significantly higher in patients with local versus systemic disease. Moreover, IL-8 expression was significantly associated to overall post-operative survival. Finally, all chemokines were strongly induced by IL-1alpha in the colon cancer cell lines tested, indicating a potential link to inflammatory processes. CONCLUSION: In accordance with earlier findings, we report here a significantly increased expression of GRO-2, GRO-3, and IL-8 in colon carcinoma as compared to normal tissue. Furthermore, GRO-3 was related to metastasis formation, and IL-8 was associated with survival, suggesting a potential predictive power of these markers.

90: Cell biology international, 2010 Feb 16, 23(3)
Effect of vascular endothelial growth factor (VEGF) on expressions of interleukin-8 (IL-8), IL-1ss and their receptors in bovine theca cells.

[Abstract]Cytokines such as vascular endothelial growth factor (VEGF) and interleukins are involved in follicular development in the mammalian ovary. The aim of the present study is to examine the transcript of the IL-8 and IL-1 differs during follicular development and the relationships between IL-8, IL-1 and VEGF in the theca cells is still unknown. We first examined the gene expression of IL-8, IL-1ss, CXCR1, and IL-1R1 in the theca cells of pre-selection (PRF) and post-selection follicles (POF) from the bovine ovary. Expression of IL-8 and CXCR1 genes were observed in POF, whereas expression of IL-1ss and IL-1R1 genes was observed in both follicles. Secondary, we examined the effects of VEGF on the expression of IL-8, IL-1ss and their receptors genes in cultured bovine theca cells. Messenger RNA (mRNA) expression was quantified by using real-time PCR methods. VEGF stimulate the expression of IL-8 and CXCR1 mRNA. However, VEGF down regulate the expression of CXCR2 mRNA during culture period. Expression of IL-1ss and IL-1R1 mRNA was induced in the cultured theca cells at 48h. Our data demonstrates that VEGF stimulated the expression of the IL-8, and CXCR1 genes and that CXCR2 expression was suppressed by VEGF, suggesting a follicle stage-dependent expression pattern for IL-8 system. Furthermore, our results suggest that the transcription system for CXCR genes may have different pathways by VEGF stimulation in bovine theca cells. Taken together, our data suggested that VEGF is associated with the IL system in the theca cells in bovine ovary.

91: Infection and immunity, 2010 Feb 9, 243(1)
The K5 capsule of Escherichia coli strain Nissle 1917 is important in stimulating expression of TLR5, CD14, MyD88 and TRIF together with the induction of IL-8 expression via the MAPK-pathway in epithelial cells.

[Abstract]Escherichia coli strain Nissle 1917, that has been widely used as a probiotic for the treatment of inflammatory bowel disorders, expresses a K5 capsule, expression of which is often associated with extra-intestinal and urinary tract isolates of E. coli. Previously it had been shown that the expression of a K5 capsule by Nissle 1917 was important in mediating interactions with epithelial cells and the extent of chemokine expression. In this paper we show that infection with Nissle 1917 induces expression of Toll Like Receptor (TLR) 4 and TLR5 in Caco-2 cells and that maximal induction of TLR5 required the K5 capsule. In addition, purified K5 polysaccharide was capable of inducing expression of TLR5 and mCD14 and potentiated the activity of both TLR4 and TLR5 agonists to increase the pro-inflammatory response. Infection with Nissle 1917 also increased expression of the adaptor molecules MyD88 and TRIF that was K5 capsule dependent. By Western blot analysis it was possible to show induction of interleukin-8 by Nissle 1917 was predominantly through the MAP kinase pathway and that expression of the K5 capsule was important for activation of the MAP-kinase pathway. This paper provides new information on the function of the K5 capsule in mediating interactions between Nissle 1917 and epithelial cells and the mechanisms that underlie the probiotic properties of Nissle 1917.

92: Pediatrics international : official journal of the Japan Pediatric Society, 2010 Feb 3, 243(1)
The association of development of chronic lung disease of newborns with neonatal colonization of Ureaplasma and Interleukin-8 level of cord blood.

[Abstract]Abstract Aims: To investigate the association of Chronic Lung Disease (CLD), neonatal Ureaplasma colonization, and the Interleukin-8 (IL-8) level of cord blood in preterm infants. Methods: In 77 infants of <32 weeks, the relationship among the IL-8 level of cord blood, neonatal colonization of Ureaplasma, histological chorioamnionitis (CAM), and development of CLD was studied. Results: Five infants died and 29 infants developed CLD. The CLD group had significantly lower gestation (mean +/- SD: 26.6 +/- 1.8 weeks) compared with the infants without CLD (28.9 +/- 1.9 p < 0.0001). Logistic analysis showed that the development of CLD was associated with gestational age (odds ratio 0.5, 95%CI 0.4-0.8) and Ureaplasma colonization (4.1, 1.2-14.4). And Ureaplasma colonization was associated with CAM (6.5, 1.8-23.5), no respiratory distress syndrome (6.2, 1.3-30.5), and development of CLD (4.0, 1.1-15.3). Elevated IL-8 level of cord blood of 100pg/ml and higher was associated with females and the isolation of microorganisms (49.4, 4.6-525). Conclusion: The development of CLD defined by oxygen requirement at 36 weeks was associated with neonatal Ureaplasma colonization but not with IL-8 of cord blood. The elevated IL-8 of cord blood was associated with neonatal microorganisms isolation.

93: Reproduction (Cambridge, England), 2010 Feb 4, 316(4)
Interleukin-8 (CXCL8) stimulates trophoblast cell migration and invasion by increasing levels of MMP-2 and MMP-9 and integrins {alpha}5 and {beta}1.

[Abstract]Interleukin-8 (IL-8/CXCL8) is present in decidua and trophoblast, which also expresses the IL-8 receptors, CXCR1 and CXCR2. IL-8 was shown to stimulate trophoblast migration. MMP-2, MMP-9, and integrins alpha5beta1 and alpha1beta1 were found to play important roles in trophoblast invasion. We hypothesized that IL-8 would increase this cell migration and invasion by HTR-8/SVneo cells, through the activity of metalloproteinases (MMPs) and integrins. Isolated first trimester of pregnancy cytotrophoblast (CTB) and HTR-8/SVneo cell line were used. Migration was studied by monolayer wounding test and invasion by Matrigel invasion test. The effects of IL-8 on MMPs and integrin subunit expression were determined on HTR-8/SVneo cells by gelatin zymography and Western blot respectively. The results obtained showed that exogenous IL-8 stimulated HTR-8/SVneo cell migration and invasion. MMP-2 and MMP-9 levels were stimulated to 182% (p<0.01) and 134% (p<0.01) respectively. Integrin alpha5 expression was increased to 119% (p<0.05) and beta1 to 173% (p<0.001) of control values. The data obtained show for the first time the sensitivity of the HTR-8/SVneo cells, in addition to isolated first trimester cytotrophoblast, to IL-8. Exogenous IL-8/CXCL8 increased trophoblast cell migration and invasion, which may be partly attributable to stimulation of MMP-2 and MMP-9 levels and an increase in integrins. HTR-8/SVneo cell viability and proliferation were also increased.

94: Pancreatology : official journal of the International Association of Pancreatology (IAP) ... [et al.], 2010 Jan 21, 9(6)
Diagnostic Accuracy of Interleukin-6 and Interleukin-8 in Predicting Severe Acute Pancreatitis: A Meta-Analysis.

[Abstract]Objectives: The early identification of patients at risk for severe acute pancreatitis (SAP) is crucial. Serum markers of disease severity have been assessed including interleukin (IL)-6 and IL-8; however, their predictive accuracy has varied significantly across studies. We conducted a meta-analysis to assess the accuracy of IL-6 and IL-8 at predicting SAP. Methods: We identified relevant published articles and calculated pooled sensitivities, specificities and likelihood ratios using the random-effect model. We included values for days 1, 2 and 3 of presentation for IL-6 and for days 1 and 2 for IL-8. We also constructed summary receiver-operating curves and assessed the area under the curve (AUC) and the diagnostic odds ratios (DORs) as measures of diagnostic accuracy. Results: For IL-6, we included 7 reports for day 1 and 4 reports for days 2 and 3. For IL-8, we analyzed 5 studies for day 1 and 4 for day 2. The pooled IL-6 sensitivities ranged between 81.0 and 83.6% and specificities between 75.6 and 85.3% with positive likelihood ratios of 3.43, 4.90 and 4.40 for days 1, 2 and 3, respectively. The IL-8 pooled sensitivities were 65.8 and 70.9% with specificities of 66.5 and 91.3% for days 1 and 2 with positive likelihood ratios of 1.96 and 8.15. The IL-6 AUCs were 0.75, 0.88 and 0.85 for days 1, 2 and 3. The IL-8 AUCs were 0.73 and 0.91 for days 1 and 2. The DOR for IL-6 was higher than that of IL-8 on day 1. Conclusion: IL-6 and IL-8 seem to perform at an acceptable level in predicting SAP. Larger confirmatory studies formally comparing this performance with that of more commonly used markers are needed. and IAP.

95: Carcinogenesis, 2010 Jan 27, 51(2)
Phospholipase D signaling pathway is involved in lung cancer-derived IL-8 increased osteoclastogenesis.

[Abstract]Bone is a frequent target of lung cancer metastasis, which is associated with significant morbidity and a dismal prognosis. This study analyzed the soluble factors secreted by lung cancer cells which are responsible for increasing osteoclast differentiation. Addition of recombinant human interleukin-8 (rhIL-8), present in large amounts in A549-condition medium (CM) and NCI-H460-CM, mimicked the inductive effect of A549-CM and NCI-H460-CM on osteoclastogenesis. In contrast, depletion of IL-8 from A549-CM and NCI-H460-CM decreased the osteoclastogenesis-inductive properties of A549-CM and NCI-H460-CM. Induction of osteoclast differentiation by lung cancer-derived CM and rhIL-8 was associated with increased phospholipase D (PLD) activation, and the activations of protein kinase C (PKC)alpha /betaII, ERK1/2 and AKT/the mammalian target of rapamycin (mTOR). Blocking PLD by a specific inhibitor significantly decreased osteoclast formation by inhibiting PKCs activation and subsequently attenuating the phosphorylation of ERK1/2. PLD inhibitor also completely decreased AKT and mTOR phosphorylation, whereas phosphatidyl-inositol-3-kinase (PI3K) inhibitor only partially decreased mTOR phosphorylation, suggesting that mTOR activation by PLD is through both a PI3K/AKT-dependent and independent manner. In addition, blocking AKT and ERK1/2 by a specific inhibitor also suppressed lung cancer-derived CM and rhIL-8-induced osteoclast differentiation. Moreover, treatment of PBMCs with sera from invasive lung cancer patients increased the formation of osteoclasts. Our study suggests that IL-8 or IL-8-mediated PLD/PKC/ERK1/2 or PLD/AKT signaling is an attractive therapeutic target for osteolytic bone metastases in lung cancer patients.

96: Cancer research, 2010 Jan 26, 51(2)
Interleukin-8 Mediates Resistance to Antiangiogenic Agent Sunitinib in Renal Cell Carcinoma.

[Abstract]The broad spectrum kinase inhibitor sunitinib is a first-line therapy for advanced clear cell renal cell carcinoma (ccRCC), a deadly form of kidney cancer. Unfortunately, most patients develop sunitinib resistance and progressive disease after about 1 year of treatment. In this study, we evaluated the mechanisms of resistance to sunitinib to identify the potential tactics to overcome it. Xenograft models were generated that mimicked clinical resistance to sunitinib. Higher microvessel density was found in sunitinib-resistant tumors, indicating that an escape from antiangiogenesis occurred. Notably, escape coincided with increased secretion of interleukin-8 (IL-8) from tumors into the plasma, and coadministration of an IL-8 neutralizing antibody resensitized tumors to sunitinib treatment. In patients who were refractory to sunitinib treatment, IL-8 expression was elevated in ccRCC tumors, supporting the concept that IL-8 levels might predict clinical response to sunitinib. Our results reveal IL-8 as an important contributor to sunitinib resistance in ccRCC and a candidate therapeutic target to reverse acquired or intrinsic resistance to sunitinib in this malignancy. Cancer Res; 70(3); 1063-71.

97: Immunology, 2010 Jan 22, 51(2)
Evaluation of localized and systemic immune responses in cutaneous leishmaniasis caused by Leishmania tropica: interleukin-8, monocyte chemotactic protein-1 and nitric oxide are major regulatory factors.

[Abstract]Summary We have established Leishmania tropica as the causative agent of cutaneous leishmaniasis (CL) in the region of India where the disease is endemic. The association between localized and circulating levels of immune-determinants in CL patients was evaluated. Reverse transcription-polymerase chain reaction analysis revealed up-regulation of interferon-gamma (IFN-gamma), interleukin (IL)-1beta, IL-8, tumour necrosis factor-alpha (TNF-alpha), IL-10 and IL-4 in dermal lesions at the pretreatment stage (n = 31) compared with healthy controls (P < 0.001) and a significant down-regulation after treatment (n = 14, P < 0.05). The results indicated that an unfavourable clinical outcome in CL was not related to an inadequate T helper 1 (Th1) cell response, but rather to impairment in multiple immune functions. Comparative assessment of treatment regimes with rifampicin (RFM) or sodium antimony gluconate (SAG) revealed tissue cytokine levels to be significantly reduced after treatment with RFM (P < 0.005), while no significant decrease was evident in the levels of IFN-gamma, TNF-alpha and IL-10 (P > 0.05) as a result of treatment with SAG. Increased transcripts of monocyte chemoattractant protein-1 (MCP-1) (P < 0.001) and inducible nitric oxide synthase (iNOS) (P < 0.05) were evident before treatment in tissue lesions and remained high after treatment. Immunohistochemistry demonstrated strong expression of myeloperoxidase (MPO) and IL-8, and moderate expression of iNOS in dermal lesions. The expression levels of IL-8, MCP-1 and nitric oxide (NO) were high in patient sera before treatment, as determined using cytokine bead array and enzyme-linked immunosorbent assay (ELISA). At the post-treatment stage, the serum IL-8 levels had decreased; however, the levels of MCP-1 and NO remained high. These data suggest that IL-8 is an effector immune-determinant in the progression of CL, whereas NO facilitates the parasite killing by macrophages via MCP-1-mediated stimulation.

98: The Journal of pharmacology and experimental therapeutics, 2010 Jan 20, 51(2)
MAP-kinase phosphatase-1 negatively regulates the expression of interleukin-6, interleukin-8 and cyclooxygenase-2 in A549 human lung epithelial cells.

[Abstract]Mitogen-activated protein kinase phosphatase-1 (MKP-1) is a protein phosphatase that regulates the activity of p38 MAP kinase and JNK and, to lesser extent, p42/44 ERK. Studies with MKP-1(-/-) mice show that MKP-1 is a regulating factor suppressing excessive cytokine production and inflammatory response. The data on the role of MKP-1 in the regulation of inflammatory gene expression in human cells is much more limited. In the present study, we investigated the effect of MKP-1 on the expression of IL-6, IL-8 and COX-2 in response to stimulation with cytokines (TNF, IL-1beta and IFNgamma; 10 ng/ml each) in A549 human lung epithelial cells. Cytokines enhanced p38 and JNK phosphorylation and MKP-1 expression. p38 MAP kinase inhibitors SB202190 and BIRB 796 inhibited cytokine-induced phosphorylation of p38 substrate MK2 and expression of IL-6, IL-8 and COX-2. An aminopyridine-based compound JNK inhibitor VIII inhibited phosphorylation of a JNK substrate c-Jun, but did not have any effect on IL-6, IL-8 or COX-2 expression. Down-regulation of MKP-1 with siRNA enhanced p38 and JNK phosphorylation, and increased IL-6, IL-8 and COX-2 expression in A549 cells. In conclusion, cytokine-induced MKP-1 expression was found to negatively regulate p38 phosphorylation and the expression of IL-6, IL-8 and COX-2 in human pulmonary epithelial cells. Our results suggest that MKP-1 is an important negative regulator of inflammatory gene expression in human pulmonary epithelial cells, and compounds that enhance MKP-1 may have anti-inflammatory effects and control inflammatory response in the human lung.

99: Pediatric nephrology (Berlin, Germany), 2010 Jan 19, 51(2)
Urinary levels of interleukin-6 and interleukin-8 in patients with vesicoureteral reflux and renal parenchymal scar.

[Abstract]The objective of this study was to assess the urine levels of interleukin-6 (IL-6) and interleukin-8 (IL-8) as noninvasive markers of vesicoureteral reflux (VUR) and renal parenchymal scarring (RPS) in children in the absence of a recent urinary tract infection (UTI) episode. Urine concentrations of IL-6 and IL-8 in 114 children aged 1 month to 16 years were evaluated. The children were divided into four groups: group 1, 26 children with VUR and RPS; group 2, 27 children with VUR without RPS; group 3, 34 children with RPS without VUR, group 4, 27 children without VUR and RPS, as the control group. After the first assessment, the children were divided into four larger groups for comparison purposes: group A (groups 1+2), 53 children with VUR; group B (groups 3+4), 61 children without VUR; group C (groups 1+3), 60 children with RPS; group D (groups 2+4), 54 children without RPS. Urinary IL-6 and IL-8 concentrations were determined. To avoid dilution effects and to the standardize samples, urinary levels of IL-6 and IL-8 were expressed as the ratio of cytokine to urinary creatinine (pg/mg). The median urine IL-6/creatinine was significantly higher in patients with VUR than in those without VUR (5.72 vs. 3.73). In patients with VUR, there was a significant but rather weak correlation between IL-6/creatinine concentrations and the reflux grade (p < 0.05, R = 0.305). The median urine IL-8/creatinine was significantly higher in patients with RPS than in those without RPS (43.12 vs. 16.36). In patients with RPS, there was a significant but rather weak correlation between IL-8/creatinine concentrations and the renal scar grade (p < 0.05, R = 0.251). The results of this study provide preliminary evidence that children with VUR have a high urine IL-6 concentration, whereas children with RPS have a high urine IL-8 concentration.

100: Journal of clinical immunology, 2010 Jan 14, 126(2)
Interleukin-32gamma Enhances the Production of IL-6 and IL-8 in Fibroblast-Like Synoviocytes Via Erk1/2 Activation.

[Abstract]INTRODUCTION: In the present study, we examined the effect of the pro-inflammatory cytokine IL-32gamma, the most biologically active isoform, and its related molecules in fibroblast-like synoviocytes (FLS). MATERIALS AND METHODS: FLS were isolated from synovial tissues of rheumatoid arthritis (RA) patients. The secretion and expression of IL-6 and IL-8 were examined by ELISA and real-time PCR, and the activation of signaling molecules was evaluated by Western blot, electrophoretic mobility shift assay (EMSA), real-time PCR, and siRNA transfection. RESULTS: By IL-32gamma stimulation in RA FLS, the expressions of IL-6 and IL-8 were increased significantly, and the phosphorylated Erk1/2 and AP-1 were expressed prominently in Western blot and EMSA. In the Erk1/2 inhibited cells, IL-32gamma stimulation did not increase the mRNA expression of IL-6 and IL-8. CONCLUSION: Our results suggest that IL-32gamma stimulation can induce the production of IL-6 and IL-8 from RA FLS via Erk1/2 activation.

101: The FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2010 Jan 11, 126(2)
Proteinase-activated receptors induce interleukin-8 expression by intestinal epithelial cells through ERK/RSK90 activation and histone acetylation.

[Abstract]Proteinase-activated receptors (PARs) are involved in both inflammation and tumorigenesis in epithelial cells. Interleukin (IL)-8 is a potent chemoattractant and is also involved in angiogenesis. The molecular mechanism whereby PARs induce epithelial IL-8 expression is not known. In HT-29 colonic epithelial cells, PAR1 or PAR2 agonists stimulated the expression of IL-8 through a NF-kappaB-dependent pathway without inducing IkappaB degradation and disassociation of IkappaB from NF-kappaB. Further studies revealed that PAR activation induced the phosphorylation of p65 at Ser-276 in the nucleus, which increased the recruitment of histone acetyltransferase (HAT) p300 to p50. Inhibition of ERK activation completely blocked PAR-induced IL-8 expression, phosphorylation of p65 and HAT activity. We also demonstrated that RSK p90 was the downstream kinase that mediated ERK-induced nuclear p65 phosphorylation. In conclusion, activation of either PAR1 or PAR2 stimulated the transcriptional up-regulation of IL-8 in HT-29 colonic epithelial cells through a pathway that involved ERK/RSK p90, NF-kappaB phosphorylation, and HAT activity. These studies provide evidence of a new role for serine proteinases and PARs in the regulation of gene expression in colonic inflammation and tumorigenesis.-Wang, H., Moreau, F., Hirota, C. L., MacNaughton, W. K. Proteinase-activated receptors induce interleukin-8 expression by intestinal epithelial cells through ERK/RSK90 activation and histone acetylation.

102: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology, 2010 Jan, 26(1)
[Relationship of IL-6 and IL-8 secretion in epithelial ovarian cancer cell lines with their sensitivity to tamoxifen as well as MAPK, Akt and estrogen receptor phosphorylation.]

[Abstract]AIM: To investigate the relationship of IL-6 and IL-8 secretion in four epithelial ovarian cancer cell lines (A2780, CAOV-3, SKOV-3 and ES-2) with their sensitivity to tamoxifen (TAM) as well as MAPK, Akt and estrogen receptor (ER) phosphorylation, and to explore the mechanism of endocrine therapy resistance caused by IL-6 and IL-8 in ovarian cancer cells. METHODS: Reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA) were performed to analyze the expression of IL-6 and IL-8. MTT assay was carried out to examine the response of ovarian cancer cells to TAM. Western blot was used to detect phosphorylated MAPK, Akt and ER. RESULTS: Except A2780 cells, three other ovarian cancer cells constitutively expressed IL-6 and IL-8. The mRNA levels of IL-6 and IL-8 correlated with their protein levels in four ovarian cancer cells. The four ovarian cancer cells showed different response to TAM. A2780 cells was the most responsive, whereas CAOV-3, SKOV-3 and ES-2 cells were TAM-resistant to a different degree. There was a notable difference in phosphorylated MAPK, Akt and ER (serine 118 and 167) among the four ovarian cancer cells. CONCLUSION: Autocrine production of IL-6 and IL-8 in epithelial ovarian cancer cell lines is inversely associated with cell response to TAM, and positively associated with phosphorylated MAPK, Akt and ER.

103: Comparative hepatology, 2010 Jan 5, 9(1)
Serum levels of soluble Fas, soluble tumor necrosis factor-receptor II, interleukin-2 receptor and interleukin-8 as early predictors of hepatocellular carcinoma in Egyptian patients with hepatitis C virus genotype-4.

[Abstract]ABSTRACT: BACKGROUND: Liver disease progression from chronic hepatitis C virus (HCV) infection to hepatocellular carcinoma (HCC) is associated with an imbalance between T-helper 1 and T-helper 2 cytokines. Evaluation of cytokines as possible candidate biomarkers for prediction of HCC was performed using soluble Fas(sFas), soluble tumor necrosis factor receptor-II (sTNFR-II), interleukin-2 receptor (IL-2R) and interleukin-8 (IL-8). RESULTS: The following patients were recruited: 79 with HCV infection, 30 with HCC, 32 with chronic liver disease associated with elevated liver enzyme levels (with or without cirrhosis) in addition to 17 with chronic HCV with persistent normal alanine aminotransferase levels (PNALT). Nine normal persons negative either for HCV or for hepatitis B virus were included as a control group. All persons were tested for sFas, sTNFR-II, IL-2R and IL-8 in their serum by quantitative ELISA. HCC patients had higher levels of liver enzymes but lower log-HCV titer when compared to the other groups. HCC patients had also significantly higher levels of sFas, sTNFR-II and IL-2R and significantly lower levels of IL-8 when compared to the other groups. Exclusion of HCC among patients having PNALT could be predicted with 90 % sensitivity and 70.6 % specificity when sTNFR-II is [greater than or equal to] 389 pg/ml or IL-8 is < 290 pg/ml. CONCLUSIONS: Serum TNFR-II, IL-2Ralpha and IL-8, may be used as combined markers in HCV-infected cases for patients at high risk of developing HCC; further studies, however, are mandatory to check these findings before their application at the population level.

104: Cytokine, 2009 Dec 29, 30(6)
Cytokines in bronchoalveolar lavage fluid and serum of lung cancer patients during radiotherapy - Association of interleukin-8 and VEGF with survival.

[Abstract]Radiotherapy (RT) produces oxidative stress and local inflammation. This study aimed at clarifying the role of different cytokines (VEGF, TNFalpha, IL-1beta, IL-6, IL-8, IL-12 and IL-18) in bronchoalveolar lavage (BAL) fluid and in the serum of lung cancer patients at baseline and during RT. Bronchoscopy and BAL were performed on 36 lung cancer patients and 36 controls for diagnosis; patients receiving RT had a second bronchoscopy during RT. Serum samples were obtained during RT and three months after RT. In this study lung cancer patients had higher levels of serum and BAL fluid IL-6 and serum IL-8 compared to controls (p<0.001, p=0.039 and p=0.030, respectively). RT caused a significant increase of BAL fluid IL-6 (p=0.037). There were no significant associations between baseline cytokine levels and adverse events or response to treatment. Higher baseline serum and BAL fluid IL-8 and serum VEGF levels (p=0.036, p=0.027 and p=0.014, respectively) were associated with shorter survival. This study shows that lung cancer is associated with upregulation of IL-6 and IL-8. The increase of BAL fluid IL-6 during RT might be attributed to enhanced RT-related oxidative stress or increased cell death. Serum and BAL fluid IL-8 and serum VEGF might have a prognostic role in survival of lung cancer.

105: International journal of oncology, 2010 Feb, 36(2)
Xanthohumol, a prenylated chalcone derived from hops, inhibits proliferation, migration and interleukin-8 expression of hepatocellular carcinoma cells.

[Abstract]Xanthohumol, the major prenylated chalcone found in hops, is well known to exert anti-cancer effects, but information regarding the impact on hepatocellular carcinoma (HCC) cells and potential adverse effects on non-tumorous hepatocytes is limited. Here, we show that xanthohumol at a concentration of 25 microM induced apoptosis in two HCC cell lines (HepG2 and Huh7). Furthermore, xanthohumol repressed proliferation and migration, as well as TNF induced NF-kappaB activity and interleukin-8 expression in both cell lines at even lower concentrations. In contrast, xanthohumol concentrations up to 100 microM did not affect viability of primary human hepatocytes in vitro. In summary, our data showed that xanthohumol can ameliorate different pro-tumorigenic mechanisms known to promote HCC progression, indicating its potential as promising therapeutic agent that selectively affects cancer cells.

106: Cellular microbiology, 2009 Dec 21, 30(6)
HIF-1alpha mediates the induction of IL8 and VEGF expression on infection with Afa/Dr Diffusely Adhering E. coli and promotes EMT-like behaviour.

[Abstract]Abstract Microbes regulate a large panel of intracellular signalling events that can promote inflammation and/or enhance tumour progression. Indeed, it has been shown that infection of human intestinal cells with the Afa/Dr diffusely adhering E. coli C1845 strain induces expression of pro-angiogenic and pro-inflammatory genes. Here, we demonstrate that exposure of cryptic-like intestinal epithelial cells to C1845 bacteria induces HIF-1alpha protein levels. This effect depends on the binding of F1845 adhesin to the membrane associated DAF receptor that initiates signalling cascades promoting translational mechanisms. Indeed, inhibition of MAPK and PI-3K decreases HIF-1alpha protein levels and blocks C1845-induced phosphorylation of the ribosomal S6 protein. Using RNA interference we show that bacteria-induced HIF-1alpha regulates the expression of IL8, VEGF and Twist1, thereby pointing to a role for HIF-1 in angiogenesis and inflammation. In addition, infection correlates with a loss of E-cadherin and cytokeratin 18 and a rise in fibronectin, suggesting that bacteria may induce an epithelial to mesenchymal transition-like phenotype. Since HIF-1alpha silencing results in reversion of bacteria-induced EMT markers, we speculate that HIF-1alpha plays a key role linking bacterial infection to angiogenesis, inflammation and some aspects of cancer initiation.

107: Neuro endocrinology letters, 2009 Dec 30, 30(6)
Value of amniotic fluid interleukin-8 for the prediction of histological chorioamnionitis in preterm premature rupture of membranes.

[Abstract]Objective: To determine whether amniotic fluid levels of interleukin-8 (IL-8) are of value in the antenatal diagnosis of acute histological chorioamnionitis (HCA) in preterm premature rupture of membranes (PPROM). Setting: Department of Obstetrics and Gynaecology, Charles University, Medical School and University Hradec Kralove, Czech Republic Methods: We compared amniotic fluid IL-8 levels in twenty-nine pregnant women with preterm premature rupture of membranes between 24th and 36th gestational weeks with presence and absence acute histological chorioamnionitis or/and microbial invasion in the amniotic cavity using nonparametric tests (Mann-Whitney test), given the non-normal distribution of analyte. Comparisons of proportions were performed with Shapiro-Wilk normality test. Results: Patients with HCA had a significantly higher median amniotic fluid IL-8 concentration than patients without the histological signs of chorioamnionitis (1867 pg/mL, 826- 5577 versus 1045 pg/mL, 60-4133, p= 0.013). Patients with MIAC had a significantly higher median amniotic fluid level than patients without invasion (1888 pg/mL, 519-5577 versus 1225 pg/mL, 60-2766, p= 0,017). Women with HCA and MIAC had a significantly higher median amniotic fluid IL-8 level than women without histological signs of chorioamnionitis and microbial invasion (3117 pg/mL, 826-5577 versus 1468 pg/mL, 394-2766, p=0,034) Conclusion: HCA or/and MIAC are associated with a significant increase of amniotic fluid interleukin-8 levels. Amniotic fluid IL-8 seems to be a marker of intraamniotic inflammation.

108: International archives of allergy and immunology, 2009 Dec 16, 152(2)
Regulation of CXCR/IL-8 in Human Airway Epithelial Cells.

[Abstract]Background: Severe asthma is characterized by neutrophilic inflammation and high levels of interleukin (IL)-8. Airway epithelial cells play a pivotal role in the pathogenesis and chronicity of asthma. The objective of this work was to determine whether CXC receptors were involved in human small airway epithelial cell (SAEC) activity by incubating them with IL-8; the investigation also included a proteomic approach. Methods: IL-6 and intercellular adhesion molecule-1 (ICAM-1) were assessed by ELISA and flow cytometry, respectively. CXCR-1 and CXCR-2 receptor mRNA and protein expressions were analyzed by RT-PCR, immunocytochemistry and flow cytometry. Cells were incubated with different concentrations (0-100 ng/ml) of IL-8. The involvement of both receptors was assessed using specific antibodies. Results: Only the CXCR-1 receptor was expressed in SAECs. IL-8 (50 ng/ml, 12 h) induced the release of IL-6 and had no effect on ICAM-1. Supernatants analyzed by surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF MS) showed very weak differences in peptide profiles. Interestingly, 4,820-m/z peptide release was detected in the presence of IL-8 and abolished by CXCR-1 antibody. Discussion: The present study illustrated the fact that IL-8 mediated by CXCR-1 increased IL-6. We also highlight the usefulness of SELDI ProteinChip technology to confirm the potential variation of peptide profile. Moreover, we were able to detect the 4,820-m/z peptide secreted in vitro by human airway epithelial cells induced by IL-8 via CXCR-1 receptor. Determination of the protein secretion profile in response to inflammatory stimuli could be an important therapeutic strategy in severe asthma.

109: Human reproduction (Oxford, England), 2009 Dec 13, 152(2)
Signal mechanisms of vascular endothelial growth factor and interleukin-8 in ovarian hyperstimulation syndrome: dopamine targets their common pathways.

[Abstract]BACKGROUND Ovarian hyperstimulation syndrome (OHSS) is a serious complication of ovarian stimulation with massive ascites, pleural effusion and hemoconcentration. The pathophysiological signal mechanisms of OHSS are still unclear and merit further investigation. METHODS Various angiogenic cytokines of follicular fluid and ascites of patients with risk of OHSS were measured, and examined for inducing endothelial permeability. These include vascular endothelial growth factor (VEGF), interleukin (IL)-6, IL-8, basic fibroblast growth factor, tumor necrosis factor-alpha, IL-1alpha, IL-1beta and platelet-derived growth factor. We explore the molecular signal pathways of major contributing cytokines in granulosa-lutein cells and endothelial cells possibly involved in OHSS. RESULTS Neutralizing antibodies of VEGF or IL-8 significantly decreased follicular fluid- and ascites-induced endothelial permeability. Human chorionic gonadotrophin induced VEGF secretion of granulosa-lutein cells through the Sp1 and CREB dependent pathways. IL-8 activated CXCR1/2 of endothelial cells leading to VEGF receptor (VEGFR)-2 transactivation. Both VEGF and IL-8 of follicular fluid enhanced endothelial permeability via VEGFR-2-mediated Rho/Rock activation, actin polymerization and phosphorylations of VE-cadherin and occludin, resulting in opening of adherens junctions and tight junctions. Dopamine (2 microM) inhibited follicular fluid-induced VEGFR-2 signals and endothelial permeability, without diminishing migration and tube formation. CONCLUSIONS Our results suggest that VEGF and IL-8 secreted from corpora luteae may play major roles in OHSS. Delineation of signal pathways would be helpful for treatment. Dopamine may block VEGF- and IL-8-induced endothelial permeability by inhibiting common VEGFR-2 dependent signals.

110: The Journal of general virology, 2009 Dec 9, 284(50)
Human papillomavirus 5 and 8 E6 downregulate interleukin-8 secretion in primary human keratinocytes.

[Abstract]Human papillomaviruses (HPVs) of genus beta appear to be involved in the early stages of skin cancer development because both the prevalence and viral load are higher in precancerous actinic keratoses than in skin cancers. IL-8 is an inflammatory cytokine, which serves to alert the surrounding tissue after UV-induced damage. We examined the effects of the E2, E6 and E7 proteins of HPV8 and the E6 proteins of various HPV genotypes on IL-8 secretion from primary keratinocytes. HPV5 and HPV8 E6 showed the highest downregulation of basal IL-8 secretion. HPV8-E6 also negatively modulated IL-8 mRNA expression and protein secretion upon UVB irradiation. The downregulation of IL-8 in actinic keratoses may weaken the response to UV-induced damage and thus favour the accumulation of UVB induced mutations.

111: Journal of leukocyte biology, 2009 Dec 9, 284(50)
The murine IL-8 homologues KC, MIP-2, and LIX are found in endothelial cytoplasmic granules but not in Weibel-Palade bodies.

[Abstract]Rapid translocation of P-selectin from WPB to the surface of endothelial cells is crucial for early neutrophil recruitment to acute inflammatory lesions. Likewise, the chemokine CXCL8/IL-8 is sorted to WPB in human endothelial cells, but little is known about its functional importance in lack of a suitable animal model. Here, we explored the distribution of the functional IL-8 homologues CXCL1/KC, CXCL2/MIP-2, and CXCL5-6/LIX in resting and inflamed murine vessels by confocal microscopy and paired immunostaining with markers of WPB, discovering that these chemokines did not localize to WPB but displayed a granular pattern in a subset in healthy skin compatible with sorting to the type 2 endothelial compartment for regulated secretion. Moreover, all chemokines colocalized with VWF and P-selectin in platelets, suggesting that their storage in platelet alpha-granules might represent an alternative source of rapidly available, neutrophil-recruiting chemokines. In conclusion, WPB appear not to be involved in regulated secretion of chemokines in the mouse, and instead, the possible existence of type 2 granules and the role of platelets in rapid leukocyte adhesion deserve further attention.

112: Cytokine, 2009 Dec 14, 152(2)
The expression of IL-8 and IL-8 receptors in pancreatic adenocarcinomas and pancreatic neuroendocrine tumours.

[Abstract]BACKGROUND: Inflammatory mediators influence tumour progression. IL-8 has been shown to have pro-angiogenic, mitogenic and motogenic effects and several studies have demonstrated the expression of IL-8 by various human pancreatic cancer cell lines. Methods: The expression of IL-8 and IL-8 receptors was studied in 52 pancreatic adenocarcinomas and 52 pancreatic neuroendocrine tumours using immunohistochemistry. The expression of IL-8 and IL-8 receptors was also assessed in eight pancreatic adenocarcinomas and seven neuroendocrine tumours in comparison to normal pancreatic tissue using real time quantitative PCR (qRT-PCR). Results: Immunohistochemical analysis of the expression of IL-8, IL-8RA and IL-8RB in 52 pancreatic adenocarcinomas demonstrated expression in 25%, 75% and 79% of pancreatic adenocarcinomas, respectively. There was no statistically significant correlation between expression and tumour grade and stage for any of the three antigens. IL-8, IL-8RA and IL-8RB expression was detected in 21%, 63% and 92% of 52 pancreatic neuroendocrine tumours. There was no statistically significant correlation between expression and tumour grade for any of the three antigens. Using qRT-PCR, the expression of each of IL-8, IL-8RA and IL-8RB mRNA was increased in 75% of pancreatic adenocarcinomas. IL-8, IL-8RA and IL-8RB mRNA expression was also increased in 57%, 43% and 29% of pancreatic neuroendocrine tumours. Quantitatively, there was a significant increase in expression level of IL-8 in tumours of both types in comparison to normal pancreatic tissue (38.5-fold in adenocarcinomas and 43.9-fold in neuroendocrine tumours). There was also increased expression of IL-8RA in both tumour types, with higher levels in adenocarcinomas, 2.7-fold and neuroendocrine tumours, 1.7-fold. IL-8RB was slightly increased in adenocarcinomas in comparison to normal pancreas (1.4-fold), but the expression was decreased in neuroendocrine tumours compared with normal pancreas (0.9-fold). Conclusion: This is the first study to show that IL-8 and IL-8 receptors are upregulated in both pancreatic adenocarcinomas and neuroendocrine tumours, and indicate this signalling pathway may modulate tumour behaviour through autocrine and/or paracrine loops.

113: Translational research : the journal of laboratory and clinical medicine, 2010 Jan, 155(1)
Systemic and airway inflammation in sleep apnea and obesity: the role of ICAM-1 and IL-8.

[Abstract]The recurrent hypoxic stress that characterizes obstructive sleep apnea (OSA) seems to play a role in the increased adherence of neutrophils to endothelial cells as well as in the resulting migration of the former to the inflamed area. Intercellular adhesion molecule 1 (ICAM-1) and interleukin (IL)-8 are markers widely used in OSA studies to investigate inflammation. The aim of this study was to measure ICAM-1 and IL-8 levels in the breath condensate and in the plasma and inflammatory cells in the induced sputum of 12 obese OSA (OO) patients, 10 nonobese OSA (NOO) patients, 10 obese non-OSA (ONO) subjects, and 8 healthy subjects (HS) using a specific enzyme immunoassay (EIA) kit. A significant increase in both plasma and exhaled IL-8 and ICAM concentrations and percentage neutrophils was observed in the induced sputum of obese OSA patients, non-obese OSA patients, and obese non-OSA subjects compared with healthy subjects. However, although these inflammatory markers were found to follow an upward trend in obese OSA patients no difference was observed in both either non-obese OSA patients and obese non-OSA subjects. Finally, a significant positive correlation was found to occur among IL-8, ICAM-1, and sputum neutrophils, as well as across the apnea-hypopnoea index (AHI), TST 90%, body mass index (BMI), and neck circumference. The data obtained confirm the occurrence of an ICAM- and IL-8-mediated neutrophilic airway inflammation in both OSA and obese patients. The degree of inflammation, which seems to worsen in cases of comorbidity (OSA and obesity), is likely to be responsible for the increased risk of developing cardiovascular events observed in these subjects, and therefore, it deserves to be elucidated even more.

114: Journal of leukocyte biology, 2009 Dec 7, 284(50)
Integrin-mediated inhibition of interleukin-8 secretion from human neutrophils by collagen type I.

[Abstract]The function of neutrophils in the inflammatory response is modulated by contact with ECM proteins. We have now investigated the effect of collagen type I on secretion of the cytokine IL-8 by human neutrophils in vitro. Collagen type I inhibited the secretion of IL-8 from neutrophils maintained under basal conditions or stimulated with fMLF. This effect was accompanied by down-regulation of IL-8 mRNA, and it appeared to be specific to collagen type I among ECM proteins, in that it was not observed with fibronectin or laminin. The inhibitory effect of collagen type I on IL-8 secretion was dependent on collagen concentration and cell density. It was also abolished in the presence of antibodies to integrin alpha2beta1 but was not affected by antibodies to integrin alpha5beta1 or beta4. Our results thus suggest that collagen type I inhibits the secretion of IL-8 by human neutrophils in a selective manner and that this effect is mediated by the interaction of collagen with integrin alpha2beta1.

115: Veterinary immunology and immunopathology, 2009 Nov 18, 284(47)
Stimulated expression of TNF-alpha and IL-8, but not of lingual antimicrobial peptide reflects the concentration of pathogens contacting bovine mammary epithelial cells.

[Abstract]We examined if and how mammary epithelial cells (MECs) calibrate and confine the intensity of an inflammatory response elicited by different concentrations of mastitis pathogens. Therefore we quantified in primary bovine MEC the effect of different E. coli pathogen concentrations upon the abundance of mRNA molecules encoding factors of immune defence. Induced synthesis of the mRNAs encoding tumor necrosis factor alpha and interleukin 8 both clearly correlated with the E. coli dose 1h after stimulation. Also the decay rate of those mRNAs reflected the pathogen load. The higher the concentration of E. coli, the faster and stronger was the up regulation and also the subsequent degradation of those particular mRNA species. Modulation of the mRNA concentration of tristetraprolin, a factor crucially involved in the mRNA degradation, followed the same pattern. In contrast, extent and kinetics of increasing the mRNA concentrations of serum amyloid A3 and lingual antimicrobial peptide were almost independent of the pathogen dose. We show that MEC perceive the information about the different pathogen concentrations and convert this signal into a calibrated synthesis of pro-inflammatory cytokines. We suggest that selective degradation of the mRNA molecules encoding those inflammatory cytokines contributes significantly to prevent an overshooting immune response in the udder.

116: Experimental cell research, 2009 Nov 24, 47(2-3)
Protein kinase C zeta regulates interleukin-8-mediated stromal-derived factor-1 expression and migration of human mesenchymal stromal cells.

[Abstract]Mesenchymal stromal cells (MSCs) are bone marrow-derived cells with multipotent differentiation capability that are mobilized into the circulation in response to injury and localize to areas of tissue damage including solid tumors. They have the capacity to adopt a phenotype similar to carcinoma-associated fibroblasts (CAFs) and, like CAFs, promote tumor growth. The molecular communication between tumor cells and MSCs has not been well defined. However, MSCs have increased expression of the chemokine stromal-derived factor 1 (SDF-1) when exposed to conditioned medium from tumor cells. Additionally, SDF-1 has been shown to be important in the promotion of tumor growth by CAFs. These data suggest that the SDF-1 signaling axis is a key feature of the tumor microenvironment. In this report, we demonstrate that interleukin 8 (IL-8) induces an increase in SDF-1 expression by MSCs. The increase in SDF-1 expression in response to IL-8 is mediated by the activation of the protein kinase C (PKC) zeta isoform. In a functional assay, activation of PKC is required for in vitro MSC migration in response to tumor conditioned medium. These results indicate that IL-8-mediated SDF-1 production by MSCs requires PKC zeta activation. This signaling pathway provides insight into possible molecular targets for cancer therapy aimed at disrupting the interaction between components of the tumor microenvironment.

117: Biochemical and biophysical research communications, 2009 Nov 20, 6(4)
Bacteria-Derived Hydrogen Sulfide Promotes IL-8 Production from Epithelial Cells.

[Abstract]Hydrogen sulfide (H(2)S), a volatile sulfur compound, is implicated as a cause of inflammation, especially when it is produced by bacteria colonizing gastrointestinal organs. However, it is unclear if H(2)S produced by periodontal pathogens affects the inflammatory responses mediated by oral/gingival epithelial cells. Therefore, the aims of this study were 1) to compare the in vitro production of H(2)S among 14 strains of oral bacteria and 2) to evaluate the effects of H(2)S on inflammatory response induced in host oral/gingival epithelial cells. P. gingivalis (Pg) produced the most H(2)S in culture, which, in turn, resulted in the promotion of proinflammatory cytokine IL-8 from both gingival and oral epithelial cells. The up-regulation of IL-8 expression was reproduced by the exogenously applied H(2)S. Furthermore, the mutant strains of Pg that do not produce major soluble virulent factors, i.e. gingipains, still showed the production of H(2)S, as well as the promotion of epithelial IL-8 production, which was abrogated by H(2)S scavenging reagents. These results demonstrated that Pg produces a concentration of H2S capable of up-regulating IL-8 expression induced in gingival and oral epithelial cells, revealing a possible mechanism that may promote the inflammation in periodontal disease.

118: Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research, 2009 Nov 24, 6(4)
Association of Interleukin-8 Gene Polymorphism With Cachexia From Patients With Gastric Cancer.

[Abstract]Interleukin (IL)-8 has been implicated in a wide range of diseases. The polymorphism of IL-8 gene, which may affect the production level of the cytokine, may be associated with cancer cachexia. To test this hypothesis, we investigated the potential influence of the polymorphisms of the IL-8 gene, -251 A/T and +781 C/T, on susceptibility to cachexia from patients with gastric cancer in a Chinese population, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). A significantly increased frequency of +781 T allele was noted in patients with cachexia (OR = 1.765, 95% CI: 1.192-2.615, P = 0.004). The +781 TT genotypes were observed to be associated with a significantly increased risk of cachexia (OR = 2.156, 95% CI: 1.056-4.400, P = 0.033), and the difference was enhanced beyond the level of statistical significance when logistic regression was applied (OR = 3.500, 95% CI: 1.406-8.710, P = 0.007). Haplotype analysis revealed that A(251)T(781) haplotype (defined by SNPs at positions -251 and +781) was associated with a significantly increased risk of cachexia as compared with the T(251)C(781) haplotype (OR = 1.69; 95% CI: 1.08-2.62; P = 0.022). These results suggest that the genetic polymorphisms of proinflammatory cytokine IL-8 may contribute to the pathogenesis of cachexia in gastric cancer patients.

119: Toxicology and applied pharmacology, 2009 Nov 12, 200(10)
Differential transcriptional regulation of IL-8 expression by human airway epithelial cells exposed to diesel exhaust particles.

[Abstract]Exposure to Diesel Exhaust Particles (DEP) induces inflammatory signaling characterized by MAP kinase-mediated activation of NFkB and AP-1 in vitro and in bronchial biopsies obtained from human subjects exposed to DEP. NFkB and AP-1 activation results in the upregulation of genes involved in promoting inflammation in airway epithelial cells, a principal target of inhaled DEP. IL-8 is a proinflammatory chemokine expressed by the airway epithelium in response to environmental pollutants. The mechanism by which DEP exposure induces IL-8 expression is not well understood. In the current study, we sought to determine whether DEP with varying organic content induces IL-8 expression in lung epithelial cells, as well as, to develop a method to rapidly evaluate the upstream mechanism(s) by which DEP induces IL-8 expression. Exposure to DEP with varying organic content differentially induced IL-8 expression and IL-8 promoter activity human airway epithelial cells. Mutational analysis of the IL-8 promoter was also performed using recombinant human cell lines expressing reporters linked to the mutated promoters. Treatment with a low organic-containing DEP stimulated IL-8 expression by a mechanism that is predominantly NFkB-dependent. In contrast, exposure to high organic-containing DEP induced IL-8 expression independently of NFkB through a mechanism that requires AP-1 activity. Our study reveals that exposure to DEP of varying organic content induces proinflammatory gene expression through multiple specific mechanisms in human airway epithelial cells. The approaches used in the present study demonstrate the utility of a promoter-reporter assay ensemble for identifying transcriptional pathways activated by pollutant exposure.

120: Endocrinology, 2009 Nov 11, 200(10)
Lysophosphatidic Acid Up-Regulates Expression of Growth-Regulated Oncogene-{alpha}, Interleukin-8, and Monocyte Chemoattractant Protein-1 in Human First-Trimester Trophoblasts: Possible Roles in Angiogenesis and Immune Regulation.

[Abstract]The serum lysophospholipase D activity and production of lysophosphatidic acid (LPA) increase in women with pregnancy. The effects of LPA on human placenta tissue remained unclear. We investigate the expression of LPA receptors and function of LPA in human first-trimester trophoblasts. Normal villous trophoblasts were obtained from termination of first-trimester gestation. We examined the expression of LPA receptors in primary culture of trophoblasts and the tissue. The effects of LPA on the expressions of chemokines of trophoblasts were examined using RT-PCR and enzyme immunoassay. We delineate signal pathways of LPA-inducing relevant chemokines in trophoblasts. The secretory chemokines were tested for angiogenic function using human endometrial microvascular endothelial cells and for immunological chemotaxis using decidual natural killer cells and THP-1 monocytes. The results revealed the expression of LPA1 receptors in trophoblast cells. LPA enhanced growth-regulated oncogene (GRO)-alpha, IL-8 and monocyte chemoattractant protein (MCP)-1 expressions in a time- and dose-dependent manner. Mechanistic dissection disclosed that LPA functioned mainly via the LPA1 receptor, Gi protein, various signal mediators of ERK, protein kinase C, p38, Akt, and c-Jun N-terminal kinase, and nuclear factor-kappaB pathways to secrete these chemokines. LPA-induced IL-8 protein secretion of trophoblasts enhanced permeability, migration, proliferation, and capillary tube formation of human endometrial microvascular endothelial cells. LPA-induced GRO-alpha and MCP-1 incited chemotaxis of natural killer cells and monocytes. We demonstrate that LPA mediates trophoblast cells to produce GRO-alpha, IL-8, and MCP-1 via LPA1 receptors and nuclear factor-kappaB-dependent signal pathways. Through LPA-induced chemokine production, human first-trimester trophoblast cells may regulate angiogenesis and innate immune system in early pregnancy.

121: Molecular and cellular biochemistry, 2009 Oct 24, 68(1)
Enteroaggregative Escherichia coli infection induces IL-8 production via activation of mitogen-activated protein kinases and the transcription factors NF-kappaB and AP-1 in INT-407 cells.

[Abstract]Enteroaggregative Escherichia coli (EAEC) is emerging as a cause of acute and persistent diarrhea in developing countries. An important feature of EAEC pathogenesis is the induction of profound inflammatory response in the intestinal epithelium. In this article, we have shown that EAEC-induced activation of mitogen-activated protein kinases (MAPK) (ERK-1/2, JNK and p38MAPK) in cultured human intestinal epithelial cells (INT-407) leads to the induction of DNA-binding activity of NF-kappaB and AP-1, resulting in IL-8 production. Plasmid-cured EAEC could also activate the MAPK and the transcription factors leading to IL-8 secretion, but to a lesser extent than that of wild-type EAEC. Further, pretreatment of these cells with the highly specific MEK inhibitor (PD 098059), the JNK inhibitor (SP 600125), and the p38MAPK inhibitor (SB 203580) resulted in inhibition of the IL-8 secretion by EAEC (wild type as well as plasmid cured)-infected INT-407 cells. These findings demonstrate that the inflammatory response induced by EAEC may be due to the specific stimulation of MAPK signaling pathways leading to nuclear responses. To our knowledge, this is the first article regarding the detailed mechanism of IL-8 secretion from the EAEC-infected human intestinal epithelial cell line.

122: Experimental dermatology, 2009 Nov 2, 200(10)
Increased mast cell expression of PAR-2 in skin inflammatory diseases and release of IL-8 upon PAR-2 activation.

[Abstract]Please cite this paper as: Increased mast cell expression of PAR-2 in skin inflammatory diseases and release of IL-8 upon PAR-2 activation. Experimental Dermatology 2009.Abstract: Mast cells are increasingly present in the lesional skin of chronic skin inflammatory diseases including psoriasis and basal cell carcinoma (BCC). It has previously been shown that proteinase-activated receptor (PAR)-2 is expressed by mast cells, and tryptase is a potent activator of this receptor. In this study, skin biopsies from both healthy-looking and lesional skin of patients with psoriasis and superficial spreading BCC were collected and the expression of PAR-2 immunoreactivity in tryptase-positive mast cells was analysed. PAR-2 expression was confirmed in vitro in different mast cell populations. Cord-blood derived mast cells (CBMC) were stimulated with a PAR-2 activating peptide, 2-furoyl-LIGRLO-NH(2). Consequently, IL-8 and histamine production was analysed in the supernatants. We observed a significant increase in the percentage of mast cells expressing PAR-2 in the lesional skin of psoriasis and BCC patients compared with the healthy-looking skin. HMC-1.2, LAD-2 and CBMC mast cells all expressed PAR-2 both intracellularly and on the cell surface. CBMC activation with the PAR-2 activating peptide resulted in an increased secretion of IL-8, but no histamine release was observed. Furthermore, both PAR-2 and IL-8 were co-localized to the same tryptase-positive mast cells in the lesional BCC skin. These results show that mast cells express increased levels of PAR-2 in chronic skin inflammation. Also, mast cells can be activated by a PAR-2 agonist to secrete IL-8, a chemokine which can contribute to the progress of inflammation.

123: Respiration; international review of thoracic diseases, 2009 Nov 3, 200(10)
Ibuprofen Modulates NF-kappaB Activity but Not IL-8 Production in Cystic Fibrosis Respiratory Epithelial Cells.

[Abstract]Background: High-dose ibuprofen is clinically effective in cystic fibrosis (CF); however, its molecular mechanisms are poorly understood. Objective: To test the hypothesis that clinically relevant concentrations of ibuprofen suppress activation of nuclear factor (NF)-kappaB and thus down-regulate stimulated interleukin (IL)-8 production in CF respiratory epithelial cells. Methods: The majority of experiments were conducted in CFTE29o- cells (F508del-mutated CF transmembrane regulator, CFTR). Key experiments were confirmed in CFBE41o- cells (F508del-mutated CFTR) and 1HAEo- cells (wild-type CFTR). NF-kappaB and IL-8 were stimulated with tumour necrosis factor (TNF)-alpha or IL-1beta. NF-kappaB and IL-8 suppression by ibuprofen (480 muM) was compared to dexamethasone (5 nM). Results: Both TNF-alpha and IL-1beta activated NF-kappaB and stimulated IL-8 production. Both ibuprofen and dexamethasone demonstrated comparably modest suppression of NF-kappaB transcriptional activity. However, ibuprofen had no effect on stimulated IL-8 mRNA and protein. By contrast, dexamethasone significantly down-regulated stimulated IL-8 mRNA and protein. Conclusions: The present data do not support the hypothesis that ibuprofen down-regulates IL-8 production in response to TNF-alpha and IL-1beta in CF respiratory epithelium. Suppression of NF-kappaB transcriptional activity does not discriminate between anti-inflammatory drugs with or without effects on IL-8 production. We speculate that NF-kappaB-independent mechanisms may be responsible for anti-IL-8 effects of dexamethasone.

124: Biological & pharmaceutical bulletin, 2009 Nov, 32(11)
Effect of resveratrol on helicobacter pylori-induced interleukin-8 secretion, reactive oxygen species generation and morphological changes in human gastric epithelial cells.

[Abstract]Inflammatory cytokine interleukin-8 (IL-8) and reactive oxygen species (ROS) overexpressed in the gastric mucosa when exposed to Helicobacter pylori, defined as a class I carcinogen. Moreover, infection with H. pylori leads to morphological changes in co-cultured cells known as hummingbird phenomenon along with increased motility. Resveratrol, a highly abundant polyphenol in red grapes, has shown anti-inflammatory, anti-cancer, cardioprotective and neuroprotective activities. However, the effect of resveratrol in H. pylori-infected cells has not been investigated. The present study was, therefore, aimed to evaluate the effect of resveratrol on the induction of IL-8, ROS and hummingbird morphology in H. pylori-infected gastric epithelial cells. The non-toxic concentration of resveratrol for both H. pylori and epithelial cells was determined by brucella broth dilution method and DNA fragmentation assay. The non-toxic resveratrol (< or =100 microM) treatment did not demonstrate any inhibitory effect against H. pylori adhesion to gastric epithelial cells. However, preincubation of the cells with 75 and 100 muM of resveratrol significantly (p<0.05 and p<0.01 respectively) inhibited the secretion of IL-8 from H. pylori-infected cells. In addition, resveratrol pretreatment at 1-100 muM suppressed H. pylori-induced ROS generation in a concentration dependent manner. Moreover, H. pylori-initiated morphological changes were markedly blocked by resveratrol. Hence, resveratrol can be considered as a potential candidate against various H. pylori related gastric pathogenic processes.

125: Biochemical pharmacology, 2009 Oct 26, 49(11)
IL-8 production by macrophages is synergistically enhanced when cigarette smoke is combined with TNF-(

[Abstract]Macrophages are key inflammatory cells in chronic obstructive pulmonary disease (COPD). The pathophysiology of cigarette smoke-induced lung emphysema is complex but there is a clear role for reactive oxygen species (ROS, such as peroxynitrite), tumor necrosis factor (TNF-() and interleukin (IL)-8. We investigated whether TNF-( or cigarette smoke medium (CSM) alone or in combination induces the production of IL-8 by human macrophages or monocyte lymphoma U937. CSM and TNF-( induces a dose- and time-dependent increase in IL-8 production. Interestingly, when sub-threshold concentrations of CSM and TNF-( were co-incubated, a 1500% increase in IL-8 production was observed compared to either of the compounds alone. Similar results were obtained with TNF-( and the peroxynitrite donor SIN-1. Moreover, the overproduction of IL-8 was associated with an enhanced increase in the translocation of NF-(B and an enhanced decrease in glutathione levels. Pre-incubation of the cells with antioxidants, such as N-acetyl-L-cysteine, prevented the overproduction of IL-8 and activation of NF-(B. In conclusion, CSM exposure of macrophages up-regulates the expression and the production of IL-8 via reactive oxygen species and NF-(B activation. Moreover, CSM dramatically enhances the production of IL-8 in combination with TNF-(. Based upon the strong synergistic action, a combination therapy directed against ROS and TNF-( could be a new approach to stop the progression in lung damage during emphysema.

126: Experimental biology and medicine (Maywood, N.J.), 2009 Nov, 234(11)
The IL-8 production by streptococcal pyrogenic exotoxin B.

[Abstract]We have previously identified alpha(v)beta(3) and Fas as receptors for the streptococcal pyrogenic exotoxin B (SPE B), and G308S, a mutant of SPE B with RSD motif, which interacts with Fas only. This study aims to evaluate how SPE B interacts with cells to induce the production of IL-8. Our results showed that following exposure to SPE B or G308S, the levels of IL-8 protein and mRNA were increased and the increase was inhibited by the addition of anti-Fas antibody, suggesting that the increased production of IL-8 by SPE B is mediated through Fas receptor. In the presence of G308S, the association of FADD and procaspase 8, and activation of NF-kappaB were also detected. The application of siRNA of FADD and of procaspase 8 could inhibit the NF-kappaB activity. The proteolytic activity of caspase 8 was required for the NF-kappaB activity. Further studies showed that G308S could increase the phosphorylation of ERK and the translocation of NF-kappaB into the nucleus, and the inhibition of ERK phosphorylation decreased the IL-8 production, mRNA expression and activation of NF-kappaB. In addition, siRNA of procaspase 8 could inhibit the G308S-induced cleavage of MEKK1, binding of MEKK1 to caspase 8, activation of ERK and the NF-kappaB activity. Taken together, the production of IL-8 by SPE B in A549 cells is mediated by Fas, and followed by the activation of FADD, caspase 8, MEKK1, ERK and NF-kappaB.

127: Journal of clinical neuroscience : official journal of the Neurosurgical Society of Australasia, 2009 Oct 19, 28(42)
Detection of MCP-1 and IL-8 in the serum and cerebrospinal fluid of a child with Miller Fisher syndrome.

[Abstract]We describe an 11-year-old female patient who presented with a 7-day history of diplopia and difficulty walking. On examination she had ataxia, areflexia and ophthalmoplegia, and a diagnosis of Miller Fisher syndrome (MFS) was made after the exclusion of other conditions. Monocyte chemotactic protein-1 (MCP-1) and interleukin (IL)-8 chemokine levels in the cerebrospinal fluid (CSF) were significantly increased in the acute phase. We believe MFS in this patient was due to both peripheral nervous system dysfunction, and central nervous system (CNS) involvement. The cause of MFS in this patient was suggested by the localized chemokine production in the CSF. The high expression of MCP-1 and IL-8 chemokines suggest that macrophages and T cells may stimulate inflammation of the CNS in MFS.

128: Inflammation, 2009 Oct 20, 28(42)
Association of Interleukin-8 Gene Polymorphisms and Haplotypes with Oral Lichen Planus in a Chinese Population.

[Abstract]Interleukin-8 (IL-8), a CXC chemokine with multiple biological functions, plays an important role in the pathogenesis of oral lichen planus (OLP). The aim of this study was to investigate the association of single nucleotide polymorphisms (SNPs) of IL-8 gene with OLP in a Chinese population. Four SNPs of the IL-8 gene at positions -845 T/C (rs2227532), -738 T/A, -251 A/T (rs4073) and +781 C/T (rs2227306) were analyzed in 109 patients with OLP and 101 normal controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The data revealed that the -251 AA genotype and -251 A allele frequency was significantly lower in the erosive OLP (eOLP) group than in the control group (P = 0.012 and P = 0.031, respectively). Haplotype analysis revealed that the -251 A/+781 C haplotype frequency was lower in the eOLP group than in the control group (P = 0.029) while the -251 T/+781 C haplotype frequency was higher in the eOLP patients than in the healthy controls (P = 0.028). The study suggests that the IL-8 polymorphisms may be associated with the severity of OLP in this Chinese cohort.

129: The Journal of biological chemistry, 2009 Oct 16, 28(42)
Targeted knockdown of EGR-1 inhibits IL-8 production and IL-8 mediated invasion of prostate cancer cells through suppressing EGR-1/NF-kappaB synergy.

[Abstract]IL-8 produced by prostate cancer cells may be responsible for the androgen-independent growth of advanced prostate cancers. Accumulating evidence from microarray analyses and animal genetic models highlights the central involvement of the transcription factor early growth response-1 (EGR-1) in prostate carcinoma progression. It is unknown, however, whether knockdown of EGR-1 inhibits IL-8 production and IL-8-mediated tumor metastasis. Here we show that EGR-1 knockdown by a specific shRNA-Egr1 inhibited gene transcription and production of IL-8 by human prostate cancer cell line DU145. Conversely, enforced expression of EGR-1 in EGR-1-lacking PC3 prostate cancer cells markedly enhanced IL-8 transcription and secretion. By using a wild-type and a series of mutant IL-8 promoter luciferase constructs, we found that NF-kappaB binding site is required for EGR-1 regulation of IL-8. Furthermore, silencing EGR-1 suppressed a synergistically functional interaction between EGR-1 and NF-kappaB. Consequently, knockdown of EGR-1 inhibited IL-8 mediated tumor colony formation and invasion. Thus, targeted knockdown of EGR-1 could be an effective therapeutic approach against prostate cancer.

130: The Journal of infectious diseases, 2009 Oct 16, 28(42)
Physiologic Cold Shock Increases Adherence of Moraxella catarrhalis to and Secretion of Interleukin 8 in Human Upper Respiratory Tract Epithelial Cells.

[Abstract]Moraxella catarrhalis, a major nasopharyngeal pathogen of the human respiratory tract, is exposed to rapid and prolonged downshifts of environmental temperature when humans breathe cold air. In the present study, we show that a 26 degrees C cold shock up-regulates the expression of UspA1, a major adhesin and putative virulence factor of M. catarrhalis, by prolonging messenger RNA half-life. Cold shock promotes M. catarrhalis adherence to upper respiratory tract cells via enhanced binding to fibronectin, an extracellular matrix component that mediates bacterial attachment. Exposure of M. catarrhalis to 26 degrees C increases the outer membrane protein-mediated release of the proinflammatory cytokine interleukin 8 in pharyngeal epithelial cells. Furthermore, cold shock at 26 degrees C enhances the binding of salivary immunoglobulin A on the surface of M. catarrhalis. These data indicate that cold shock at a physiologically relevant temperature of 26 degrees C affects the nasopharyngeal host-pathogen interaction and may contribute to M. catarrhalis virulence.

131: Arteriosclerosis, thrombosis, and vascular biology, 2009 Oct 15, 28(42)
Adenosine Modulates HIF-1{alpha}, VEGF, IL-8, and Foam Cell Formation in a Human Model of Hypoxic Foam Cells.

[Abstract]OBJECTIVE: Foam cell (FC) formation by oxidized low-density lipoprotein (oxLDL) accumulation in macrophages is crucial for development of atherosclerosis. Hypoxia has been demonstrated in atherosclerosis and hypoxia-inducible factor-1 (HIF-1) has been shown to promote intraplaque angiogenesis and FC development. As hypoxia induces HIF-1alpha stabilization and adenosine (ado) accumulation, we investigated whether this nucleoside regulates HIF-1alpha in FCs. METHODS AND RESULTS: Ado, under hypoxia, stimulates HIF-1alpha accumulation by activating all adenosine receptors (ARs). HIF-1alpha modulation involved extracellular signal-regulated kinase 1/2 (ERK 1/2), p38 mitogen-activated protein kinase (p38 MAPK), and protein kinase B (Akt) phosphorylation in the case of A1, A2A, A2B, and ERK 1/2 phosphorylation in the case of A3 receptors. Ado, through the activation of A3 and A2B receptors, stimulates vascular endothelial growth factor (VEGF) secretion in a HIF-1alpha-dependent way. Furthermore, ado, through the A2B subtype, induces an increase of Interleukin-8 (IL-8) secretion in a ERK 1/2, p38, and Akt kinase-dependent but not HIF-1alpha-mediated way. Finally, ado stimulates FC formation, and this effect is strongly reduced by A3 and A2B blockers and by HIF-1alpha silencing. CONCLUSIONS: This study provides the first evidence that A3, A2B, or mixed A3/A2B antagonists may be useful to block important steps in the atherosclerotic plaque development ado-induced.

132: Proceedings of the National Academy of Sciences of the United States of America, 2009 Oct 6, 106(40)
Cell surface-bound IL-1alpha is an upstream regulator of the senescence-associated IL-6/IL-8 cytokine network.

[Abstract]Inflammation underlies most age-related diseases, including cancer, but the etiology is poorly understood. One proposed factor is the presence of senescent cells, which increase with age. The senescence response arrests the proliferation of potentially oncogenic cells, and most senescent cells secrete high levels of proinflammatory cytokines and other proteins. The complex senescence-associated secretory phenotype is likely regulated at multiple levels, most of which are unknown. We show that cell surface-bound IL-1alpha is essential for signaling the senescence-associated secretion of IL-6 and IL-8, 2 proinflammatory cytokines that also reinforce the senescence growth arrest. Senescent human fibroblasts expressed high levels of IL-1alpha mRNA, intracellular protein, and cell surface-associated protein, but secreted very little protein. An IL-1 receptor (IL1R) antagonist, neutralizing IL-1alpha antibodies, and IL-1alpha depletion by RNA interference all markedly reduced senescence-associated IL-6/IL-8 secretion. Depletion of the key IL-1R signaling component IRAK1 also suppressed this secretion, and IL-1alpha neutralizing antibodies prevented IRAK1 degradation, indicating engagement of the IL-1R signaling pathway. Furthermore, IL-1alpha depletion reduced the DNA binding activity of NF-kappaB and C/EBPbeta, which stimulate IL-6/IL-8 transcription. IL-1alpha was a general regulator of senescence-associated IL-6/IL-8 secretion because IL-1alpha blockade reduced IL-6/IL-8 secretion whether cells senesced owing to DNA damage, replicative exhaustion, oncogenic RAS, or chromatin relaxation. Furthermore, conditioned medium from IL-1alpha-depleted senescent cells markedly reduced the IL-6/IL-8-dependent invasiveness of metastatic cancer cells, indicating that IL-1alpha regulates the biological effects of these cytokines. Thus, cell surface IL-1alpha is an essential cell-autonomous regulator of the senescence-associated IL-6/IL-8 cytokine network.

133: The Journal of biological chemistry, 2009 Nov 20, 284(47)
Phosphoglucose Isomerase/Autocrine Motility Factor Promotes Melanoma Cell Migration through ERK Activation Dependent on Autocrine Production of Interleukin-8.

[Abstract]It is well known that phosphoglucose isomerase/autocrine motility factor (AMF) promotes cell migration in an autocrine manner in various tumor cells. However, it remains unclear whether certain cytokines modulate the effects of AMF on tumor cell migration. Because interleukin (IL)-8, a proinflammatory cytokine, is produced by melanoma cells and has been correlated with melanoma migration, the migratory ability of melanoma cells induced by AMF may also involve induction of IL-8 expression. In the present study, we assessed whether AMF promotes melanoma cell migration through autocrine production of IL-8. We found that AMF stimulation increased IL-8 production through up-regulation of IL-8 mRNA transcription, especially in biologically early stage melanoma cells. AMF-induced migration of these cells was inhibited by a specific neutralizing antibody against IL-8. The IL-8 production induced by AMF was mediated by the ERK1/2 pathways. These findings suggest that melanoma migration induced by AMF is mediated by autocrine production of IL-8 as a novel downstream modulator of the AMF signaling pathway.

134: Journal of biomedical science, 2009, 16(40)
Difference in the regulation of IL-8 expression induced by uropathogenic E. coli between two kinds of urinary tract epithelial cells.

[Abstract]Bacterial adherence to epithelial cells is a key virulence trait of pathogenic bacteria. The type 1 fimbriae and the P-fimbriae of uropathogenic Escherichia coli (UPEC) have both been described to be important for the establishment of urinary tract infections (UTI). To explore the interactions between the host and bacterium responsible for the different environments of UPEC invasion, we examined the effect of pH and osmolarity on UPEC strain J96 fimbrial expression, and subsequent J96-induced interleukin-8 (IL-8) expression in different uroepithelial cells. The J96 strain grown in high pH with low osmolarity condition was favorable for the expression of type 1 fimbriae; whereas J96 grown in low pH with high osmolarity condition was beneficial for P fimbriae expression. Type 1 fimbriated J96 specifically invaded bladder 5637 epithelial cells and induced IL-8 expression. On the contrary, P fimbriated J96 invaded renal 786-O epithelial cells and induced IL-8 expression effectively. Type 1 fimbriated J96-induced IL-8 induction involved the p38, as well as ERK, JNK pathways, which leads to AP-1-mediated gene expression. P fimbriated J96-induced augmentation of IL-8 expression mainly involved p38-mediated AP-1 and NF-kappaB transcriptional activation. These results indicate that different expression of fimbriae in J96 trigger differential IL-8 gene regulation pathways in different uroepithelial cells.

135: International journal of oncology, 2009 Nov, 35(5)
Cepharanthine inhibits angiogenesis and tumorigenicity of human oral squamous cell carcinoma cells by suppressing expression of vascular endothelial growth factor and interleukin-8.

[Abstract]Cepharanthine is a biscoclaurine alkaloid extracted from Stephania cepharantha Hayata, which is widely used for the treatment of many acute and chronic diseases, and can exert antitumor effects on several human cancer cell lines. However, little is known about the detailed mechanisms of the antitumor activity of Cepharanthine. In the present study, we determined whether Cepharanthine could suppress angiogenesis and growth of human oral squamous cell carcinoma (OSCC) cells in vitro and in vivo. Cepharanthine significantly inhibited expression of two major pro-angiogenic molecules, vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8), in cultured cells and in cells implanted into the subcutaneous tissue of nude mice. Also, Cepharanthine inhibited the nuclear factor-kappaB (NF-kappaB) activity in human OSCC cells in vitro and in vivo. The decreased expression of VEGF and IL-8 correlated with decreased tumor cell growth and decreased vascularization in vitro and in vivo. These findings suggest that Cepharanthine can suppress angiogenesis and growth of OSCC cells by inhibiting expression of VEGF and IL-8 involved in the blockade of NF-kappaB activity.

136: Molecular immunology, 2009 Dec, 47(2-3)
Induction of IL-8 expression by bacterial flagellin is mediated through lipid raft formation and intracellular TLR5 activation in A549 cells.

[Abstract]We investigated the mechanism for the induction of a chemokine, IL-8, by bacterial flagellins in the human alveolar type II epithelial cell line, A549. Bacterial flagellin induced expression of IL-8 mRNA and protein in dose- and time-dependent manners. IL-8 expression was inhibited by nystatin (a lipid rafts inhibitor) but not by chlorpromazine (a clathrin-coated pits inhibitor). Interestingly, Toll-like receptor 5 (TLR5) recognizing flagellins was found in the intracellular compartment of A549 but rarely on the cell surface. Flagellin-induced IL-8 expression appears to be mediated through TLR5 as determined by in vitro transient transfection experiment in HEK-293 cells expressing TLR5 using a reporter gene construct containing IL-8 promoter. IL-8 expression was attenuated by inhibitors for protein kinase C (PKC) and mitogen-activated protein (MAP) kinases. Furthermore, NF-kappaB and NF-IL6 transcription factors played an important role in the flagellin-induced IL-8 gene expression in A549 cells. Collectively, these results suggest that flagellin-induced IL-8 expression requires formation of lipid rafts, intracellular TLR activation, and subsequent activation of PKC and MAP kinases leading to the activation of the transcription factors NF-kappaB and NF-IL6 in human alveolar type II epithelial cells.

137: Digestive diseases and sciences, 2010 Jul, 55(7)
Polymorphism of IL-8 in 251 allele and gastric cancer susceptibility: a meta-analysis.

[Abstract]BACKGROUND: The relationship of gastric cancer to the presence of interleukin-8 (IL-8) 251 T/A has been reported with conflicting results. AIM: To further explore the association of IL-8 251 allele polymorphism with gastric cancer susceptibility. METHODS: We performed an extensive search of relevant studies and carried out a meta-analysis, including ten studies with 2,195 gastric cancer cases and 3,505 controls, to obtain a more precise estimate. RESULTS: The combined results based on all studies showed that the IL-8 251 allele AA genotype was a risk factor for gastric cancer [AA versus TT: odds ratio (OR) = 1.363, 95% confidence interval (CI): 1.199-1.527]. In subgroup analysis, a clear effect of AA in IL-8 251 allele was shown in Asians (AA versus TT: OR = 1.593, 95% CI: 1.013-2.173) but not in Caucasians or Mexicans. When stratified by Lauren classification, we found that the IL-8 251 allele TA and AA polymorphism was significantly associated with the diffuse type of gastric cancer (TA versus TT: OR = 1.448, 95% CI: 1.177-1.720; AA versus TT: OR = 1.586, 95% CI: 1.128-2.044). The IL-8 251 AA genotype was found to be a risk factor for cardiac gastric cancer (AA versus TT: OR = 1.840, 95% CI: 1.112-2.568) but not for noncardiac gastric cancer. CONCLUSIONS: This meta-analysis suggested that IL-8 251 allele A>T polymorphism might be a risk factor for gastric cancer.

138: Rheumatology international, 2010 Jul, 30(9)
Effects of (-)-epigallocatechin-3-gallate on cyclooxygenase 2, PGE(2), and IL-8 expression induced by IL-1beta in human synovial fibroblasts.

[Abstract]The objective of this study was to examine the effects of (-)-epigallocatechin-3-gallate (EGCG) on cyclooxygenase 2 (COX-2), prostaglandin E(2) (PGE(2)), and interleukin 8 (IL-8) expression induced by IL-1beta in human synovial fibroblasts. Cells were enzymatically isolated from synovial tissue taken from patients undergoing joint replacement surgery for osteoarthritis. Reverse transcriptase-polymerase chain reaction, immunocytochemistry, and western blotting were used to assess the COX-2 gene and protein expression with the associated mechanisms. PGE(2) and IL-8 secretion into the culture medium was assayed by enzyme-linked immunosorbent assay. COX-2 upregulation in synovial fibroblasts induced by IL-1beta was significantly suppressed by EGCG in a dose-dependent manner. PGE(2) and IL-8 secretion was also induced by IL-1beta stimulation and significantly suppressed by EGCG. The mechanism was associated with the phosphorylation of IKKbeta. EGCG may inhibit the expression of inflammatory mediators, such as COX-2, PGE(2), and IL-8, induced by IL-1beta in human synovial fibroblasts. EGCG may be of value in the treatment of synovial inflammation.

139: Cytokine, 2009 Dec, 48(3)
Increased circulating IL-8 is associated with reduced IGF-1 and related to poor metabolic control in adolescents with type 1 diabetes mellitus.

[Abstract]Background: A dysregulated growth hormone (GH)/insulin-like growth factor 1 (IGF-1) axis is well-recognized in children and adolescents with type 1 diabetes mellitus (T1DM). Decreased IGF-1 levels can also be found in chronic inflammatory diseases, while hyperglycemia promotes inflammatory cytokine production. Therefore, inflammatory cytokines may link poor metabolic control with GH/IGF-1 axis changes. This study examined the relationship between serum inflammatory cytokines and IGF-1 in adolescents (age 13-18) with TIDM in chronic poor (n=17) or favorable (n=19) glucose control. Poor control (PC) was defined as 3, consistent HbA1C>9% during the previous 2years, while favorable control (FC) was consistent levels of HbA1C<9%. Results: HbA1C (FC: 7.5+/-0.6%; PC: 10.5+/-0.9%, p<0.001) and interleukin (IL)-8 (FC: 3.7+/-4.0pg/ml; PC: 7.4+/-4.3pg/ml, p=0.01) were increased and IGF-1 (FC: 536.5+/-164.3ng/ml; PC: 408.9+/-157.1ng/ml, p=0.03) was decreased in patients with poor control compared to patients with favorable control. Moreover, IL-8 was inversely correlated with IGF-1 (r=-0.40, p=0.03) and positively correlated with HbA1C (r=0.36, p=0.03). Conclusions: In adolescents with T1DM and chronic, poor glucose control, increased serum IL-8 is associated with reduced IGF-1 suggesting a pro-inflammatory milieu that may contribute to alterations in the GH/IGF-1 axis.

140: International immunopharmacology, 2009 Dec, 9(13-14)
Serum levels of IL-8 and IL-6 in the long term pulmonary complications induced by sulfur mustard: Sardasht-Iran Cohort Study.

[Abstract]Sulfur mustard (SM) is a blistering chemical agent which has short and long term toxicity against many organs. The respiratory tract is one of the main targets, and is the most disabling long term complication of SM. Inflammatory mediators especially IL-8 and IL-6 play the primary role in the various chronic pulmonary diseases. Sardasht-Iran Cohort Study (SICS) was designed to evaluate immunological and molecular parameters in SM exposed people 20 years after exposure. In the present study, the association of the serum levels of IL-8, IL-6, C reactive protein (CRP) and rheumatoid factor (RF) with long term pulmonary involvement was evaluated. There were 348 exposed and 120 control participants. The clinical evaluations were done for all subjects and Spirometry was performed according to American Thoracic Society Criteria. Severity of pulmonary involvement was assessed by Global Initiative for chronic Obstructive Lung Disease (GOLD) classification. The serum levels of IL-8, IL-6 and RF were assessed by ELISA assay. CRP was assessed by photometric method. The serum levels of IL-8 and IL-6 significantly decreased in the SM exposed participants compared to the control group. There were no significant associations between the serum levels of IL-8 and pulmonary symptoms (chronic cough, sputum, hemoptysis, and dyspnea), pulmonary findings (crackles, rales, and wheezing) as well as spirometry parameters. IL-6 was associated with wheezing and CRP was associated with wheezing and rales in SM exposed group. We concluded the serum levels of these inflammatory mediators probably do not have any major role in pathogenesis and persistence of pulmonary complications and do not reflect the degree of severity of pulmonary involvement following SM exposure.

141: Biochemical and biophysical research communications, 2009 Nov 20, 389(3)
Mechanical stretch enhances IL-8 production in pulmonary microvascular endothelial cells.

[Abstract]In patients with acute respiratory distress syndrome, mechanical over-distension of the lung by a large tidal volume causes further damage and inflammation, called ventilator-induced lung injury (VILI), however, it is unclear how mechanical stretch affects the cellular functions or morphology in human pulmonary microvascular endothelial cells (HPMVECs). IL-8 has been proposed to play an important role in the progression of VILI by activating neutrophils. We demonstrated that HPMVECs exposed to cyclic uni-axial stretch produce IL-8 protein with p38 activation in strain- and time-dependent manners. The IL-8 synthesis was not regulated by other signal transduction pathways such as ERK1/2, JNK, or stretch-activated Ca(2+) channels. Moreover, cyclic stretch enhanced IL-6 and monocyte chemoattractant protein-1 production and reoriented cell perpendicularly to the stretch axis accompanied by actin polymerization. Taken together, IL-8 production by HPMVECs due to excessive mechanical stretch may activate neutrophilic inflammation, which leads to VILI.

142: Journal of dermatological science, 2009 Nov, 56(2)
Nicotinamide inhibits Propionibacterium acnes-induced IL-8 production in keratinocytes through the NF-kappaB and MAPK pathways.

[Abstract]BACKGROUND: Propionibacterium acnes (P. acnes) has been implicated in the inflammatory phase of acne vulgaris. It has been shown to activate interleukin-8 (IL-8) secretion by interacting with Toll-like receptor 2 (TLR-2) on the surface of keratinocytes. Nicotinamide has been shown to be an effective treatment for skin inflammation in various conditions, including acne vulgaris. OBJECTIVE: To investigate the molecular mechanisms underlying the anti-inflammatory properties of nicotinamide in keratinocytes stimulated by P. acnes. METHODS: HaCaT cells and primary keratinocyte cell lines were stimulated by P. acnes in the presence of nicotinamide. IL-8 production was monitored by ELISA on the cell culture supernatant and by qRT-PCR on total RNA extract. A luciferase reporter system assay was used to assess nicotinamide activity with the IL-8 promoter in transfected keratinocytes. We used western blotting to analyze the effect of nicotinamide on activation of the NF-kappaB and MAPK pathways. RESULTS: Nicotinamide significantly decreased IL-8 production in a dose-dependent manner, decreasing both mRNA and protein levels for this chemokine in immortalized HaCaT cells and primary keratinocytes. P. acnes-induced IL-8 promoter activation seemed to be downregulated by nicotinamide, which inhibited IkappaB degradation and the phosphorylation of ERK and JNK MAP kinases. CONCLUSION: Our results indicate that nicotinamide inhibits IL-8 production through the NF-kappaB and MAPK pathways in an in vitro keratinocytes/P. acnes model of inflammation. Keratinocytes involved in the innate immune response may be a suitable target for treatment during the early phase of inflammation.

143: International immunopharmacology, 2009 Nov, 9(12)
Modulation of expression of IL-8 gene in bronchial epithelial cells by 5-methoxypsoralen.

[Abstract]Persistent recruitment of neutrophils in the bronchi of cystic fibrosis patients contributes to airway tissue damage, suggesting the importance of intervening on the expression of the neutrophil chemokine IL-8. Extracts from plants have been investigated to select components able to reduce IL-8 expression in bronchial epithelial cells challenged with Pseudomonas aeruginosa. Extracts and purified components have been added to cells 24 h before pro-inflammatory challenge with P. aeruginosa and IL-8 transcription was quantified in the IB3-1 CF cells in vitro. P. aeruginosa-dependent IL-8 mRNA induction was increased by Argemone mexicana and Vernonia anthelmintica whereas no significant modification of transcription was observed with Aphanamixis polystachya, Lagerstroemia speciosa and Hemidesmus indicus. Finally, inhibition of IL-8 was observed with Polyalthia longifolia (IC50=200 microg/ml) and Aegle marmelos (IC50=20 microg/ml). Compounds from A. marmelos were isolated and identified by GC-MS. No significant effect was observed with butyl-p-tolyl sulphate, whereas the inhibition obtained with 6-methyl-4-chromanone concentration was accompanied by an anti-proliferative effect. On the contrary, 5-methoxypsoralen resulted in IL-8 inhibition at 10 microM concentration, without effects on cell proliferation. In synthesis, 5-methoxypsoralen can be taken into consideration to investigate mechanisms of neutrophil chemotactic signalling and for its potential application in modulating the excessive CF lung inflammation.

144: Epidemiology and infection, 2010 Apr, 138(4)
Associations of plasma IL-8 levels with Helicobacter pylori seropositivity, gastric atrophy, and IL-8 T-251A genotypes.

[Abstract]SUMMARYThere are few data on circulatory pro-inflammatory cytokine levels and cytokine gene polymorphisms in H. pylori-positive patients. A cross-sectional study was conducted to examine the effects of H. pylori infection, gastric atrophy, and the IL-8 T-251A polymorphism on plasma IL-8 levels in 98 Japanese adults. Seventy-one subjects were positive for H. pylori infection. The geometric mean of plasma IL-8 concentration was significantly higher in subjects with H. pylori infection than in those without (P=0.001). The development of atrophy was negatively associated with IL-8 levels in the H. pylori-positive subjects, although not significantly. Plasma IL-8 levels in the T/T genotype were associated with H. pylori infection and atrophy status (P=0.016). Our findings suggested that circulating IL-8 levels were associated with H. pylori infection. The effect of H. pylori infection on plasma IL-8 levels was not clearly modified by the IL-8 T-251A polymorphism.

145: Inflammatory bowel diseases, 2010 Mar, 16(3)
Effect of natural commensal-origin DNA on toll-like receptor 9 (TLR9) signaling cascade, chemokine IL-8 expression, and barrier integritiy of polarized intestinal epithelial cells.

[Abstract]BACKGROUND AND AIM:: The intestinal epithelium is constantly exposed to high levels of genetic material like bacterial DNA. Under normal physiological conditions, the intestinal epithelial monolayer as a formidable dynamic barrier with a high-polarity structure facilitates only a controlled and selective flux on components between the lumen and the underlining mucosa and even is able to facilitate structure-based macromolecules movement. The aim of this study was to test the effect of natural commensal-origin DNA on the TLR9 signaling cascade and the barrier integrity of polarized intestinal epithelial cells (IECs). METHODS:: Polarized HT-29 and T84 cells were treated with TNF-alpha in the presence or absence of DNA from Lactobacillus rhamnosus GG (LGG) and Bifidobacterium longum. TLR9 and interleukin-8 (IL-8) mRNA expression was assessed by semiquantitative and TaqMan real-time reverse-transcription polymerase chain reaction. Expression of TLR9 protein, degradation of inhibitor of kappa B alpha (IkappaBalpha), and p38 mitogen-activated protein kinase (p38 MAP) phosphorylation were assessed by Western blotting. To further reveal the role of TLR9 signaling, the TLR9 gene was silenced by siRNA. IL-8 secretion was measured by an enzyme-linked immunosorbent assay. Nuclear factor-kappa B (NF-kappaB) activity was assessed by the electrophoretic mobility shift assay (EMSA) and NF-kappaB-dependent luciferase reporter gene assays. As an indicator of tight junction formation and monolayer integrity of epithelial cell monolayers, transepithelial electrical resistance (TER) was repetitively monitored. Transmonolayer movement of natural commensal-origin DNA across monolayers was monitored using qRT-PCR and nested PCR based on bacterial 16S rRNA genes. RESULTS:: In response to apically applied natural commensal-origin DNA, polarized HT-29 and T84 cells enhanced expression of TLR9 in a specific manner, which was subsequently associated with attenuation of TNF-alpha-induced NF-kappaB activation and NF-kappaB-mediated IL-8 expression. TLR9 silencing abolished this inhibitory effect. Apically applied LGG DNA attenuated TNF-alpha-enhanced NF-kappaB activity by reducing IkappaBalpha degradation and p38 phosphorylation. LGG DNA did not decrease the TER but rather diminished the TNF-alpha-induced TER reduction. Translocation of natural commensal-origin DNA into basolateral compartments did not occur under tested conditions. CONCLUSIONS:: Our study indicates that TLR9 signaling mediates, at least in part, the anti-inflammatory effects of natural commensal-origin DNA on the gut because TLR9 silencing abolished the inhibitory effect of natural commensal-origin DNA on TNF-alpha-induced IL-8 secretion in polarized IECs. The nature of the TLR9 agonist, the polarity of cells, and the tight junction integrity of IECs has to be taken into account in order to predict the outcome of TLR9 signaling. (Inflamm Bowel Dis 2010).

146: Inflammatory bowel diseases, 2010 Apr, 16(4)
Platelet-activating factor-induced NF-kappaB activation and IL-8 production in intestinal epithelial cells are Bcl10-dependent.

[Abstract]BACKGROUND:: Platelet-activating factor (PAF), a potent proinflammatory phospholipid mediator, has been implicated in inducing intestinal inflammation in diseases such as inflammatory bowel disease (IBD) and necrotizing enterocolitis (NEC). However, its mechanisms of inducing inflammatory responses are not fully understood. Therefore, studies were designed to explore the mechanisms of PAF-induced inflammatory cascade in intestinal epithelial cells. METHODS:: Nuclear factor kappa B (NF-kappaB) activation was measured by luciferase assay and enzyme-linked immunosorbent assay (ELISA), and interleukin 8 (IL-8) production was determined by ELISA. B-cell lymphoma 10 (Bcl10), caspase recruitment domain-containing membrane-associated guanylate kinase protein 3 (CARMA3), and mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) mRNA and protein levels were assessed by real-time reverse-transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. siRNA silencing of Bcl10 was used to examine its role in PAF-induced NF-kappaB activation and IL-8 production. The promoter region of the Bcl10 gene was cloned with the PCR method and promoter activity measured by luciferase assay. RESULTS:: The adaptor protein Bcl10 appeared to play an important role in the PAF-induced inflammatory pathway in human intestinal epithelial cells. Bcl10 was required for PAF-induced IkappaBalpha phosphorylation, NF-kappaB activation, and IL-8 production in NCM460, a cell line derived from normal human colon, and Caco-2, a transformed human intestinal cell line. PAF also stimulated Bcl10 interactions with CARMA3 and MALT1, and upregulated Bcl10 expression in these cells via transcriptional regulation. CONCLUSIONS:: These findings highlight a novel PAF-induced inflammatory pathway in intestinal epithelial cells, requiring Bcl10 as a critical mediator and involving CARMA3/Bcl10/MALT1 interactions. The proinflammatory effects of PAF play prominent roles in the pathogenesis of IBD and this pathway may present important targets for intervention in chronic inflammatory diseases of the intestine. (Inflamm Bowel Dis 2009;).

147: Fish & shellfish immunology, 2009 Dec, 27(6)
Molecular evidence for the existence of two distinct IL-8 lineages of teleost CXC-chemokines.

[Abstract]Interleukin-8 (IL-8) is a CXC-type chemokine with a chemotactic activity mainly on neutrophils and plays a key role in promoting inflammation. In teleosts, several CXC-chemokines have been cloned and characterized as being IL-8-like. Phylogenetic data however indicate that the reported teleost IL-8-like chemokines are substantially remote from mammalian IL-8, forming a fish-specific clade of IL-8-like chemokines distinct from that of tetrapod IL-8. In the present study, a novel IL-8-like chemokine, designated CaIL-8, has been found in the expressed sequence tags of carp gills and identified as an orthologue of mammalian IL-8. The CaIL-8 transcript encodes 99 amino acids containing a typical CXC motif but lacks an ELR motif, as in most teleost IL-8-like chemokines. Phylogenetic tree constructed by the maximum likelihood method suggests a closer relationship of CaIL-8 with mammalian IL-8 than with other teleost CXC-chemokines reported to be IL-8-like. In a normal unstimulated carp, CaIL-8 mRNA was detected by RT-PCR only in gills, kidney, spleen, heart and peripheral blood leukocytes, in contrast to a previously reported carp IL-8-like chemokine CXCa, which shows ubiquitous basal expression. The results, taken together, are strongly indicative of the presence of two major IL-8-like lineages of CXC-chemokines in teleost.

148: Oncogene, 2009 Oct 22, 28(42)
The prolyl isomerase Pin1 regulates the NF-kappaB signaling pathway and interleukin-8 expression in glioblastoma.

[Abstract]The brain tumor glioblastoma (GBM) remains one of the most aggressive and devastating tumors despite decades of effort to find more effective treatments. A hallmark of GBM is the constitutive activation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB) signaling pathway, which regulates cell proliferation, inflammation, migration and apoptosis. The prolyl isomerase, Pin1, has been found to bind directly to the NF-kappaB protein, p65, and cause increases in NF-kappaB promoter activity in a breast cancer model. We now present evidence that this interaction occurs in GBM and that it has important consequences on NF-kappaB signaling. We demonstrate that Pin1 levels are enhanced in primary GBM tissues compared with controls, and that this difference in Pin1 expression affects the migratory capacity of GBM-derived cells. Pin1 knockdown decreases the amount of activated, phosphorylated p65 in the nucleus, resulting in inhibition of the transcriptional program of the IL-8 gene. Through the use of microarray, we also observed changes in the expression levels of other NF-kappaB regulated genes due to Pin1 knockdown. Taken together, these data suggest that Pin1 is an important regulator of NF-kappaB in GBM, and support the notion of using Pin1 as a therapeutic target in the future.

149: Pediatric research, 2009 Nov, 66(5)
Clearance of apoptotic neutrophils is diminished in cord blood monocytes and does not lead to reduced IL-8 production.

[Abstract]Phagocytosis of apoptotic cells, e.g., neutrophils, by monocytes is essential for resolution of inflammation. Delayed removal leads to secondary necrosis, perpetuating inflammation, and tissue destruction. Common histologic features in neonatal chronic inflammatory disorders are an accumulation of apoptotic cells in inflamed tissues. We hypothesized that apoptotic cell removal by monocytes is compromised in newborns. PKH-26 labeled autologous or allogeneic apoptotic neutrophils were fed to monocytes of adult donors (PBMO) and cord blood (CBMO), and phagocytic activity was analyzed by flow cytometry and confocal microscopy. Relative mRNA-expression levels of 21 surface receptors and bridging molecules relevant for apoptotic cell removal were measured, as was postphagocytic IL-8 production upon LPS-stimulation. Compared with PBMO, CBMO exhibited a significantly diminished phagocytotic competence for autologous and allogeneic apoptotic neutrophils. mRNA-expression levels of milk fat globule-EGF factor 8 and T cell immunoglobulin- and mucin-domain-containing molecule, two crucial members of the phagocytic synapse of apoptotic cell removal, were reduced in CBMO. In PBMO, interaction with autologous apoptotic neutrophils reduced LPS-induced IL-8 production whereas it was enhanced in CBMO. Our data suggest a specific defect in CBMO during clearance of apoptotic neutrophils resulting in impaired anti-inflammatory capacity.

150: Journal of lipid research, 2010 Feb, 51(2)
Surfactant lipids regulate LPS-induced interleukin-8 production in A549 lung epithelial cells by inhibiting translocation of TLR4 into lipid raft domains.

[Abstract]In addition to providing mechanical stability, growing evidence suggests that surfactant lipid components can modulate inflammatory responses in the lung. However, little is known of the molecular mechanisms involved in the immunomodulatory action of surfactant lipids. This study investigates the effect of the lipid-rich surfactant preparations Survanta, Curosurf, and the major surfactant phospholipid dipalmitoylphosphatidylcholine (DPPC) on interleukin-8 (IL-8) gene and protein expression in human A549 lung epithelial cells using immunoassay and PCR techniques. To examine potential mechanisms of the surfactant lipid effects, Toll-like receptor 4 (TLR4) expression was analyzed by flow cytometry, and membrane lipid raft domains were separated by density gradient ultracentrifugation and analyzed by immunoblotting with anti-TLR4 antibody. The lipid-rich surfactant preparations Survanta, Curosurf, and DPPC, at physiological concentrations, significantly downregulated lipopolysaccharide (LPS)-induced IL-8 expression in A549 cells both at the mRNA and protein levels. The surfactant preparations did not affect the cell surface expression of TLR4 or the binding of LPS to the cells. However, LPS treatment induced translocation of TLR4 into membrane lipid raft microdomains, and this translocation was inhibited by incubation of the cells with the surfactant lipid. This study provides important mechanistic details of the immune-modulating action of pulmonary surfactant lipids.

151: Cardiovascular research, 2009 Dec 1, 84(3)
Interleukin 8 and cardiovascular disease.

[Abstract]Since the establishment of the inflammatory basis of atherosclerosis, several pro- or anti-inflammatory agents have been examined as potential mediators of the biochemical pathways of lesion formation. Interleukin (IL)-8 was first characterized in 1987. Since then, knowledge regarding its role in leucocyte trafficking and activation has advanced rapidly, especially in the field of cardiovascular disease. In the scientific literature, there is sufficient evidence to support beyond any doubt the involvement of IL-8 in the establishment and preservation of the inflammatory micro-environment of the insulted vascular wall. However, how the information derived from in vitro studies and animal models can be applied in clinical practice has yet to be determined. In the present review, the available evidence regarding the role of IL-8 in cardiovascular disease is presented, and future perspectives are discussed.

152: American journal of respiratory cell and molecular biology, 2010 Jun, 42(6)
Impaired Interleukin-8 Chemokine Secretion by Staphylococcus aureus-Activated Epithelium and T-Cell Chemotaxis in Cystic Fibrosis.

[Abstract]Staphylococcus aureus is frequently isolated from lungs of patients with cystic fibrosis (CF). Upon lung infection with S. aureus, airway epithelial cells (AEC) produce high levels of chemokines that enhance T-cell chemotaxis. Although the number of lymphocytes is increased in the airways and bronchoalveolar lavage fluid of patients with CF, the mechanisms responsible for their accumulation and the role of S. aureus in this process are largely unknown. This study investigated early S. aureus impact on chemokine secretion by CF epithelial cells and chemotaxis of CF T cells. CF and non-CF AEC were grown in a cell culture model and apically stimulated with S. aureus. Supernatants were quantified for chemokine secretions and assayed for T-cell chemotaxis. CF AEC secreted constitutively larger amounts of IL-8, GROalpha, MIG, MIP-3beta, and MCP-1 than non-CF epithelial cells. S. aureus interaction with epithelial cells increased chemokine production by non-CF cells but had no effect on CF cells. Chemotaxis of T cells derived from patients with CF was greater than that of T cells from subjects without CF. Moreover, there were more CF T cells expressing CXCR1 as compared with non-CF T cells. Under our experimental conditions, inhibition of IL-8 or its receptor CXCR1 resulted in a considerable decrease in T-cell chemotaxis (up to 80%). These data suggest that IL-8 and its receptor CXCR1 are key players in the chemotaxis of CF T cells and could be used as targets to develop therapies for CF.

153: International journal of cancer. Journal international du cancer, 2010 Jan 15, 126(2)
Selective upregulation of interleukin-8 by human rhabdomyosarcomas in response to hypoxia: therapeutic implications.

[Abstract]Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma of adolescence and childhood. Because RMS tumors are highly vascularized, we sought to determine which factors secreted by RMS cells are crucial in stimulating angiogenesis in response to hypoxia. To address this issue, we evaluated expression of several proangiogenic factors [interleukin (IL)-8, vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF)-2, stromal-derived factor (SDF)-1, hepatocyte growth factor (HGF) and leukemia inhibitory factor (LIF)] in 8 human RMS cell lines in both normal steady-state and hypoxic conditions. We found by real-time quantitative polymerase chain reaction (RQ-PCR) and confirmed by enzyme-linked immunosorbent assay (ELISA) that from all the factors evaluated, IL-8, whose expression is very low in normoxia, had been very highly expressed and secreted by RMS cells lines during hypoxic conditions ( approximately 40-170 times). Interestingly, this upregulation was not affected by knocking down hypoxia-inducible factor (HIF)-1alpha, but was inhibited by mitogen-activated protein kinase (MAPK)p42/44 and phosphatidylinositaol 3-kinase (PI3K)/AKT pathway inhibitors. This suggests that IL-8 expression is regulated in an activating protein (AP)-1- and nuclear factor (NF)-kappaB-dependent manner. Furthermore, we found that conditioned media (CM) harvested from RMS cells exposed to hypoxia activated and stimulated chemotactic responses in human umbilical vein endothelial cells (HUVECs) and that IL-8 was responsible for hypoxia-related effects. Finally, by employing shRNA, the expression of IL-8 in human RH-30 cells was downregulated. We noticed that such RMS cells, if injected into skeletal muscles of immunodeficient mice, have a reduced ability for tumor formation. We conclude that IL-8 is a pivotal proangiogenic factor released by human RMS cells in hypoxic conditions and that the targeting of IL-8 may prove to be a novel and efficient strategy for inhibiting RMS growth.

154: The European respiratory journal : official journal of the European Society for Clinical Respiratory Physiology, 2010 Jan, 35(1)
Interleukin-8 activates coagulation and correlates with survival after talc pleurodesis.

[Abstract]The aim of our study was to investigate whether interleukin (IL)-8 activates systemic coagulation after talc pleurodesis in malignant pleural effusion (MPE), and whether levels of IL-8 in plasma are related to early death after talc pleurodesis. IL-8 and tumour necrosis factor (TNF)-alpha were measured in samples from 231 MPE patients before and after talc pleurodesis. Whole blood from 31 healthy volunteers was incubated with IL-8, TNF-alpha and thromboplastin for 3 h in vitro, and thrombin-antithrombin (TAT) levels were measured. The same stimulation of blood samples was repeated using doses of calibrated talc. Nine, 12 and 17 patients died within 7, 10 and 15 days respectively. IL-8 was elevated in 102 patients within 48 h, and thrombotic events were observed in six of those patients. Survival correlated inversely with IL-8 at 24 and 48 h, and a significant correlation was also found between IL-8 and TAT. A positive dose-dependent correlation with TAT production was observed when blood was stimulated with IL-8 in vitro. However, there was no significant response to stimulation with talc, as compared with control blood samples. IL-8 is involved in the activation of coagulation that may occur after talc pleurodesis, and might also be implicated in early death of patients with MPE.

155: Protein expression and purification, 2009 Nov, 68(1)
High level expression and purification of active recombinant human interleukin-8 in Pichia pastoris.

[Abstract]Human interleukin-8 (hIL-8) is a member of interleukin family which functions as a chemotactic factor as well as an angiogenesis mediator. Previously, a study reported that hIL-8 could be purified from inclusion bodies using a prokaryotic expression system, however, the required re-naturation step limits the recovery of fully active protein. In this study, soluble recombinant hIL-8 was expressed as a secreted protein at high level in Pichia pastoris under the control of AOX1 (alcohol oxidase 1) promoter. A simple purification strategy was established to recover rhIL-8 from the fermentation supernatant. The process includes precipitation with 80% saturation ammonium sulfate and CM Sepharose ion exchange chromatography, yielding 30 mg/L purified rhIL-8 at over 95% purity. The obtained rhIL-8 displays high specific activity, stimulating the migration of mouse neutrophils at concentrations as low as 0.25 ng/mL. Our results demonstrate that P. pastoris expression system is an efficient tool for large-scale manufacture of active recombinant hIL-8 for various applications.

156: Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases, 2010 Mar, 16(3)
Interleukin-8 production by polymorphonuclear leukocytes from patients with chronic infected leg ulcers treated with Lactobacillus plantarum.

[Abstract]Bacterial infection impairs the healing process, promoting the chronicity of inflammation and wounds. Because antibiotics fail to eradicate bacteria, especially in biofilm form, new therapeutic modalities may be required. In the present study, the effectiveness of bacteriotherapy with Lactobacillus plantarum on infected chronic venous ulcers was investigated and its effects on interleukin (IL)-8 production by cells from the ulcer bed and neutrophils isolated from peripheral blood that were previously challenged in vitro with Pseudomonas aeruginosa and L. plantarum were studied. Topical application of L. plantarum culture to lesions (25-60 cm(2)) of 14 diabetic and 20 non-diabetic patients induced debridement, granulation tissue formation and total healing after 30 days in 43% diabetics and in 50% non-diabetics. No significant differences between the groups were observed. The cells from ulcer beds collected after treatment with L. plantarum for 10 days showed a decrease in the percentage of polymorphonuclear, apoptotic and necrotic cells and an enhancement of IL-8 production. IL-8 production by isolated neutrophils from these patients was compared with that in diabetics without ulcers, as well as normal subjects under basal conditions, and after infection of polymorphonuclear cells with P. aeruginosa preincubated either with or without L. plantarum. The basal values in diabetic and ulcer patients were higher than normal (p <0.001) and were increased by P. aeruginosa infection in normal, diabetics (p <0.001) and non-diabetics with ulcers (p <0.01). Preincubation with L. plantarum decreased IL-8 production in patients with ulcers non-diabetic and diabetic (p <0.001). Lactobacillus plantarum treatment reduced wound bacterial load, neutrophils, apoptotic and necrotic cells, modified IL-8 production and induced wound healing.

157: Applied biochemistry and biotechnology, 2010 Feb, 160(4)
Role of Toll-Like Receptor 3, RIG-I, and MDA5 in the Expression of Mesothelial IL-8 Induced by Viral RNA.

[Abstract]Interleukin-8 (IL-8) is a chemokine that has been shown to be a potent chemoattractant for polymorphonuclear neutrophils from the vascular compartment into the pleural space during infectious pleural effusions. Mesothelial cells express the viral receptors Toll-like receptor 3 (TLR3), RIG-I, and MDA5. Activation of these receptors by viral RNA exemplified by poly (I:C) RNA leads to a time- and dose-dependent increase of mesothelial IL-8 synthesis. To show the specific effect of viral receptors, knockdown experiments with short interfering RNA specific for TLR3, RIG-I and MDA5 were performed. This novel finding of functional expression of these viral sensors on human mesothelial cells may indicate a novel link between viral infections and mesothelial inflammation and indicates a pathophysiologic role of viral receptors in these processes.

158: The European respiratory journal : official journal of the European Society for Clinical Respiratory Physiology, 2009 Dec, 34(6)
Muscarinic M3 receptor stimulation increases cigarette smoke-induced IL-8 secretion by human airway smooth muscle cells.

[Abstract]Acetylcholine is the primary parasympathetic neurotransmitter in the airways and is known to cause bronchoconstriction and mucus secretion. Recent findings suggest that acetylcholine also regulates aspects of remodelling and inflammation through its action on muscarinic receptors. In the present study, we aimed to determine the effects of muscarinic receptor stimulation on cytokine production by human airway smooth muscle cells (primary and immortalised cell lines). The muscarinic receptor agonists carbachol and methacholine both induced modest effects on basal interleukin (IL)-8 and -6 secretion, whereas the secretion of RANTES, eotaxin, vascular endothelial growth factor-A and monocyte chemoattractant protein-1 was not affected. Secretion of IL-8 and -6 was only observed in immortalised airway smooth muscle cells that express muscarinic M3 receptors. In these cells, methacholine also significantly augmented IL-8 secretion in combination with cigarette smoke extract in a synergistic manner, whereas synergistic effects on IL-6 secretion were not significant. Muscarinic M3 receptors were the primary subtype involved in augmenting cigarette smoke extract-induced IL-8 secretion, as only tiotropium bromide and muscarinic M3 receptor subtype selective antagonists abrogated the effects of methacholine. Collectively, these results indicate that muscarinic M3 receptor stimulation augments cigarette smoke extract-induced cytokine production by airway smooth muscle. This interaction could be of importance in patients with chronic obstructive pulmonary disease.

159: Fertility and sterility, 2010 Jul, 94(2)
Interleukin-8 serum levels do not correlate with pelvic pain in patients with ovarian endometriomas.

[Abstract]OBJECTIVE: To determine whether interleukin-8 (IL-8) serum levels are correlated with pelvic pain in patients with ovarian endometriomas. DESIGN: Prospective study. SETTING: Tertiary-care university hospital. PATIENT(S): Interleukin-8 serum levels were prospectively analyzed in 51 patients (group A, asymptomatic patients or patients with mild dysmenorrhea; group B, severe dysmenorrhea and/or chronic pelvic pain and/or dyspareunia) who underwent surgery for cystic ovarian endometriosis to asses whether a correlation exists between IL-8 serum levels and pelvic pain. INTERVENTION(S): Interleukin-8 serum levels determination. MAIN OUTCOME MEASURE(S): Interleukin-8 serum levels and pelvic pain. RESULT(S): From 56 patients, five cases were ultimately excluded because the histologic diagnosis was not cystic ovarian endometriosis (2 teratomas and 3 haemorragic cysts). The mean (+/-SD) IL-8 serum levels in group A were 6.41 +/- 12.17 pg/mL and in group B were 6.52 +/- 8.73 pg/mL. CONCLUSION(S): Pain symptoms in ovarian endometriosis is not correlated with IL-8 serum levels.

160: Journal of thrombosis and thrombolysis, 2010 Jan, 29(1)
Interleukin 8 gene polymorphisms and susceptibility to restenosis after percutaneous coronary intervention.

[Abstract]Interleukin-8 is a strong mediator of inflammation and has been implicated in the biochemical pathways involved in a wide range of inflammatory diseases including atherosclerosis. We investigated the potential influence of two common functional polymorphisms of the interleukin (IL)-8 gene: -251A/T and 781C/T on susceptibility to in stent restenosis (ISR) following percutaneous coronary intervention (PCI). The hypothesis was tested by screening for the prevalence of the above polymorphisms in 201 coronary artery disease (CAD) patients subjected to PCI and presenting with symptoms or signs of recurrent ischemia. Patients were angiographically re-evaluated and formed the ISR group (n = 73) and the non-ISR group (n = 128) based on the presence or absence of ISR. One hundred and forty-seven subjects without angiographic evidence of CAD formed a reference control group (non-CAD group). A borderline statistically significant higher frequency of the TT(251)TT(781) combined genotype was observed in patients with ISR on re-evaluation compared with patients with normal follow-up angiography. The predominance of TT(251)TT(781) was independent of conventional risk factors for cardiovascular disease. Consequently, T(251)T(781) haplotype was significantly more common in the ISR group. The above observations indicate that the genetic diversity of the IL-8 gene influences patient susceptibility to ISR and suggests the implication of IL-8-mediated pathways in the process of ISR. However, the rarity of T(251)T(781) haplotype makes any clinical application of the above observations unfeasible.

161: American journal of respiratory cell and molecular biology, 2010 Jan, 42(1)
Cell-bound IL-8 increases in bronchial epithelial cells after arylsulfatase B silencing due to sequestration with chondroitin-4-sulfate.

[Abstract]The chemokine IL-8 is critically important in inflammatory processes in human tissues, and IL-8 interactions with sulfated glycosaminoglycans have been implicated in modification of inflammatory responses in bronchial epithelium. To determine the role of chondroitin-4-sulfate (C4S) in mediating effects of IL-8, we silenced the enzyme N-acetylgalactosamine-4-sulfatase (arylsulfatase B [ASB]) that removes the 4-sulfate group from C4S, in the IB3-1 and C38 bronchial epithelial cell lines and in normal primary bronchial epithelial cells. When ASB was silenced and IL-8 production stimulated by exposure to TNF-alpha, ASB activity declined by roughly 75%, cellular C4S content increased by over 7.5 microg/mg protein, cell-bound IL-8 increased by over 530 pg/mg protein, and secreted IL-8 declined by over 520 pg/mg protein in all cell lines (P < 0.001). When cell lysates were immunoprecipitated with C4S antibody after ASB silencing and TNF-alpha, the IL-8 content of the immunoprecipitate was approximately 500 pg/mg protein, indicating that most of the cell-bound IL-8 was associated with C4S. Cell fractionation demonstrated that the IL-8 content associated with the cell membranes was about twice that of the cytosolic fraction. Also, ASB appeared to localize in the cell membrane, as well as in lysosomes. Neutrophil attraction to the cell lysates increased after ASB silencing, consistent with increased attraction to the cell-bound IL-8. These findings provide evidence for the influential role of ASB and C4S in the regulation of IL-8 secretion, and suggest that changes in ASB activity and C4S content may have a significant impact on IL-8-mediated inflammatory responses.

162: Burns : journal of the International Society for Burn Injuries, 2010 May, 36(3)
Interleukin-8 is elevated in cerebrospinal fluid following high-voltage electrical injury with late-onset paraplegia suggesting neuronal damage at the microlevel as causative factor.

[Abstract]

163: Fertility and sterility, 2010 Feb, 93(2)
The human ovarian follicular fluid level of interleukin-8 is associated with follicular size and patient age.

[Abstract]OBJECTIVE: To investigate the relationship between interleukin-8 (IL-8) in the human ovarian follicle and follicular size, patient age, and fertility factors in IVF cycles. DESIGN: Prospective study. SETTING: University hospital research laboratory and infertility clinic. PATIENT(S): Women undergoing IVF with oocyte retrieval. INTERVENTION(S): Follicular fluid (FF) aspiration, oocyte isolation, FF storage, and experimental studies. MAIN OUTCOME MEASURE(S): Quantization of IL-8 by ELISAs and protein microarray; high-performance liquid chromatography (HPLC) followed by ELISA and Western blotting to evaluate alpha(2)-macroglobulin (alpha(2)M) bound IL-8; association of IL -8 to follicular size, patient age, and IVF outcomes. RESULT(S): Samples of FF from 63 patients contained an average of 629.59 pg/mL of IL-8 with 50%-70% bound to alpha(2)M. Large follicles contained higher levels of IL-8 than small follicles (937.34 vs. 86.97 pg/mL). The IL-8 concentration in the large follicles of women of young age was higher than that of older reproductive age women (1,373.61 vs. 673.29 pg/mL). There were no statistically significant associations found between IL-8 concentration and other IVF cycle factors or pregnancy outcome. CONCLUSION(S): Our findings indicate that IL-8 is present in FF, both in its free and alpha(2)M-bound state, and its concentration is correlated with follicular size and patient age.

164: Antioxidants & redox signaling, 2009 Mar 2,
HIF-1 induction attenuates Nrf2-dependent IL-8 production in human endothelial cells.

[Abstract]Through hypoxia-inducible factor 1 (HIF-1) hypoxia regulates the expression of numerous genes and is a potent inducer of angiogenesis. However, IL-8, an important angiogenic mediator, has been reported to be down-regulated by HIF-1, although the mechanisms have not been elucidated. HIF-1 was induced in human endothelial cells by hypoxia and dimethyloxaloylglycine (DMOG). Interestingly, both hypoxia and DMOG attenuated IL-8 expression and a similar effect has been obtained by adenoviral overexpression of the stable form of HIF-1alpha. Heme oxygenase-1 (HO-1) expression was also down-regulated by HIF-1 induction. This suggests similar mechanisms of regulation of IL-8 and HO-1, indicating the involvement of Nrf2, a transcription factor previously linked to hypoxia-mediated inhibition of HO-1. Indeed, HIF-1-mediated down-regulation of both IL-8 and HO-1 was associated with both lowered Nrf2 expression and induction of Bach1, a repressor of Nrf2 transcriptional activity. Accordingly, overexpression of Nrf2 reversed the inhibitory effect of HIF-1 on IL-8 and HO-1 expression. However, neither overexpression of HO-1 nor HO-1 inhibition affected IL-8 synthesis. The data indicate that HIF-1-dependent inhibition of IL-8 expression is caused by down-regulation of Nrf2. However, expression of IL-8 is independent of HO-1. Cross-talk between HIF-1 and Nrf2 may influence the outcome of anti-angiogenic therapies aimed at targeting HIF-1.

165: Respiration; international review of thoracic diseases, 2009 Mar 2,
Comparison of 8-Isoprostane and Interleukin-8 in Induced Sputum and Exhaled Breath Condensate from Asymptomatic and Symptomatic Smokers.

[Abstract]Background: Markers of airway inflammation and oxidative stress have been mainly investigated in moderate/severe chronic obstructive pulmonary disease (COPD) or during its exacerbation. They have not been compared in noninvasive specimens such as exhaled breath condensate (EBC) and induced sputum in healthy nonsymptomatic smokers or in those who have symptoms and are at risk for COPD development. Objectives: To compare the relative proportions of 2 potential COPD biomarkers, 8-isoprostane and interleukin- 8 (IL-8) in the induced sputum and EBC sampled from the same subjects: nonsmokers (n = 14), healthy smokers (n = 17) and symptomatic smokers (n = 9) who are considered to be at risk for COPD. COPD patients with acute exacerbation (n = 10) were employed as positive controls. Methods: The levels of the aforementioned biomarkers in induced sputum and EBC were investigated using commercial biochemical techniques. Results: In induced sputum, the levels of 8-isoprostane and IL-8 were at least 10-fold higher compared to EBC levels in all groups. Healthy nonsmokers had the lowest levels, and patients with exacerbation of COPD the highest levels of 8-isoprostane in induced sputum and EBC. The same observation held true for IL-8 in induced sputum. Inverse correlations with lung function parameters were observed for both mediators. Conclusions: The levels of both potential markers were clearly higher in the induced sputum than in EBC. The results point to an advantage of induced sputum over EBC for assessing the degree of airway oxidative stress and inflammation in smokers with a potential risk for COPD development.

166: American journal of reproductive immunology (New York, N.Y. : 1989), 2009 Mar, 61(3)
Effect of sulfasalazine on basal and bacteria-stimulated interleukin-8 production by endocervical epithelial cells.

[Abstract]PROBLEM: Sulfasalazine (SASP) inhibits lipopolysaccharide-induced nuclear-factor kappa B activation and interleukin-8 (IL-8) production by cultured explants of placenta, amnion and choriodecidua. Bacteria-induced IL-8 production in the cervix is a potential mechanism for premature cervical ripening that may lead to preterm birth. Our objective was to determine if SASP inhibits IL-8 production by endocervical cells stimulated with bacterial pathogens associated with preterm birth. METHOD OF STUDY: Human endocervical cells were incubated with 0-1.6 mm SASP and then stimulated with Ureaplasma parvum, Escherichia coli, or Gardnerella vaginalis. Conditioned medium was then harvested and production of IL-8 was quantified by ELISA. Viability of the cells was ascertained at the end of the experiment with the MTT-assay. RESULTS: At the highest concentration tested (1.6 mm), SASP significantly inhibited IL-8 production by cultures stimulated with E. coli (P < 0.001), U. parvum (P < 0.001), and G. vaginalis (P < 0.001). Viability of the cells, however, was significantly reduced by SASP at 0.8 and 1.6 mm in both the presence and absence of bacteria for all experiments. CONCLUSION: Although high concentrations of SASP inhibit IL-8 production by cultures of endocervical cells stimulated with pathogens associated with preterm birth, this effect may be because of toxicity of the antibiotic on the cells.

167: Carcinogenesis, 2009 Feb 20, 32(2)
Prostate stromal cells produce CXCL-1, CXCL-2, CXCL-3 and IL-8 in response to epithelia-secreted IL-1.

[Abstract]It is well accepted that tumor microenvironment is essential for tumor cells survival, cancer progression and metastasis. However, the mechanisms by which tumor cells interact with their surrounding at early stages of cancer development are largely unidentified. The aim of this study was to identify specific molecules involved in stromal-epithelial interactions which might contribute to early stages of prostate tumor formation. Here we show that conditioned medium from immortalized non-transformed prostate epithelial cells stimulated immortalized prostate stromal cells to express cancer-related molecules. Conditioned medium obtained from epithelial cells triggered stromal cells to express and secrete CXCL-1, CXCL-2, CXCL-3 and IL-8 chemokines. This effect was predominantly mediated by the cytokines of the IL-1 family secreted by the epithelial cells. Thus, prostate epithelial cells induced the secretion of pro-inflammatory and cancer promoting chemokines by prostate stromal cells. Such interactions might contribute to prostatic inflammation and progression at early stages of prostate cancer formation.

168: Pain, 2009 Feb 20, 32(2)
Upregulation of IL-6, IL-8 and CCL2 gene expression after acute inflammation: Correlation to clinical pain.

[Abstract]Tissue injury initiates a cascade of inflammatory mediators and hyperalgesic substances including prostaglandins, cytokines and chemokines. Using microarray and qRT-PCR gene expression analyses, the present study evaluated changes in gene expression of a cascade of cytokines following acute inflammation and the correlation between the changes in the gene expression level and pain intensity in the oral surgery model of tissue injury and acute pain. Tissue injury resulted in a significant upregulation in the gene expression of interleukin-6 (IL-6; 63.3-fold), IL-8 (8.1-fold), chemokine (C-C motif) ligand 2 (CCL2; 8.9-fold), chemokine (C-X-C motif) ligand 1 (CXCL1; 30.5-fold), chemokine (C-X-C motif) ligand 2 (CXCL2; 26-fold) and annexin A1 (ANXA1; 12-fold). The upregulation of IL-6 gene expression was significantly correlated to the upregulation of IL-8, CCL2, CXCL1 and CXCL2 gene expression. Interestingly, the tissue injury-induced upregulation of IL-6, IL-8 and CCL2 gene expression, was positively correlated to pain intensity at 3h post-surgery, the onset of acute inflammatory pain. However, ketorolac treatment did not have a significant effect on the gene expression of IL-6, IL-8, CCL2, CXCL2 and ANXA1 at the same time point of acute inflammation. These results demonstrate that the upregulation of IL-6, IL-8 and CCL2 gene expression contributes to the development of acute inflammation and inflammatory pain. The lack of effect of ketorolac on the expression of these gene products may be related to the ceiling analgesic effects of non-steroidal anti-inflammatory drugs.

169: Cytokine, 2009 Feb 14, 32(2)
Developmental changes in circulating IL-8/CXCL8 isoforms in neonates.

[Abstract]Interleukin-8 (IL-8/CXCL8) is widely expressed in fetal tissues although inflammatory changes are not seen. Circulating IL-8 is comprised of an endothelial-derived [ala-IL-8](77) isoform and another, more potent [ser-IL-8](72) secreted by most other cells; [ala-IL-8](77) can be converted into [ser-IL-8](72) by proteolytic removal of an N-terminal pentapeptide from [ala-IL-8](77). In this study, we show [ala-IL-8](77) is the predominant circulating isoform of IL-8 in premature neonates but not in term neonates/adults, who have [ser-IL-8](72) as the major isoform. This isoform switch from the less potent [ala-IL-8](77) to [ser-IL-8](72) correlates with a maturational increase in the neutrophil chemotactic potency of plasma IL-8. The emergence of [ser-IL-8](72) as the major isoform is likely due to increased plasma [ala-IL-8](77)-convertase activity and/or changes in the cellular sources of IL-8. Developmental changes in IL-8 isoforms may serve to minimize its inflammatory effects in the fetus and also provide a mechanism to restore its full activity after birth.


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