interferon, gamma

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

INTERFERON, GAMMA

LATEST LITERATURE

1: Gesundheitswesen (Bundesverband der Arzte des Offentlichen Gesundheitsdienstes (Germany)), 2010 Sep 2, 34(7)
Implementation of the Interferon-gamma Release Assay for Contact Tracing - A 2-Year Project in Lower Saxony, Germany.

[Abstract]B7-H1 (PD-L1) is a B7-related protein that inhibits T-cell responses. B7-H1 participates in the immunoescape of cancer cells and is also involved in the long-term persistence of leukemic cells in a mouse model of leukemia. B7-H1 can be constitutively expressed by cancer cells, but is also induced by various stimuli. Therefore, we examined the constitutive and inducible expression of B7-H1 and the consequences of this expression in human acute myeloid leukemia (AML). We analyzed B7-H1 expression in a cohort of 79 patients with AML. In addition, we studied blast cells after incubation with interferon-gamma or toll-like receptors (TLR) ligands. Finally, we evaluated functionality of cytotoxic T-cell activity against blast cells. Expression of B7-H1 upon diagnosis was high in 18% of patients. Expression of TLR2, 4 and 9 was detected in one-third of AML samples. Expression of TLR2 and TLR4 ligands or IFN-gamma induced by B7-H1 was found to protect AML cells from CTL-mediated lysis. Spontaneous B7-H1 expression was also found to be enhanced upon relapse in some patients. MEK inhibitors, including UO126 and AZD6244, reduced B7-H1 expression and restored CTL-mediated lysis of blast cells. In AML, B7-H1 expression by blasts represents a possible immune escape mechanism. The inducibility of B7-H1 expression by IFN-gamma or TLR ligands suggests that various stimuli, either produced during the immune response against leukemia cells or released by infectious microorganisms, could protect leukemic cells from T cells. The efficacy of MEK inhibitors against B7-H1-mediated inhibition of CTLs suggests a possible cancer immunotherapy strategy using targeted drugs.

2: Cancer immunology, immunotherapy : CII, 2010 Sep 4, 34(7)
In acute myeloid leukemia, B7-H1 (PD-L1) protection of blasts from cytotoxic T cells is induced by TLR ligands and interferon-gamma and can be reversed using MEK inhibitors.

[Abstract]B7-H1 (PD-L1) is a B7-related protein that inhibits T-cell responses. B7-H1 participates in the immunoescape of cancer cells and is also involved in the long-term persistence of leukemic cells in a mouse model of leukemia. B7-H1 can be constitutively expressed by cancer cells, but is also induced by various stimuli. Therefore, we examined the constitutive and inducible expression of B7-H1 and the consequences of this expression in human acute myeloid leukemia (AML). We analyzed B7-H1 expression in a cohort of 79 patients with AML. In addition, we studied blast cells after incubation with interferon-gamma or toll-like receptors (TLR) ligands. Finally, we evaluated functionality of cytotoxic T-cell activity against blast cells. Expression of B7-H1 upon diagnosis was high in 18% of patients. Expression of TLR2, 4 and 9 was detected in one-third of AML samples. Expression of TLR2 and TLR4 ligands or IFN-gamma induced by B7-H1 was found to protect AML cells from CTL-mediated lysis. Spontaneous B7-H1 expression was also found to be enhanced upon relapse in some patients. MEK inhibitors, including UO126 and AZD6244, reduced B7-H1 expression and restored CTL-mediated lysis of blast cells. In AML, B7-H1 expression by blasts represents a possible immune escape mechanism. The inducibility of B7-H1 expression by IFN-gamma or TLR ligands suggests that various stimuli, either produced during the immune response against leukemia cells or released by infectious microorganisms, could protect leukemic cells from T cells. The efficacy of MEK inhibitors against B7-H1-mediated inhibition of CTLs suggests a possible cancer immunotherapy strategy using targeted drugs.

3: Human reproduction (Oxford, England), 2010 Sep 2, 34(7)
Interferon gamma contributes to preimplantation embryonic development and to implantation site structure in NOD mice.

[Abstract]BACKGROUND Pre-eclampsia, a syndrome usually accompanied by incomplete spiral arterial modification, occurs at an increased frequency in diabetic women. Hyperglycemia in non-obese type 1 diabetic (NOD) mice impairs gestational spiral arterial remodeling despite high local levels of interferon gamma (Ifng), the triggering cytokine in mice. Pregnancies in NOD.Ifng(-/-) mice were assessed to investigate this issue. METHODS Fecundity was assessed using the breeding history, flushing of preimplantation embryos and histological and morphometric studies of implantation sites in normoglycemic (n-) and hyperglycemic (d-) females of NOD.Ifng(-/-) and NOD genotypes. RESULTS NOD.Ifng(-/-) but not NOD mice are mostly infertile. In NOD.Ifng(-/-), copulation often does not result in a post-implantation pregnancy. Defective fertilization and delayed preimplantation development limit n-NOD.Ifng(-/-) fertility, and both mechanisms are exacerbated by hyperglycemia. At mid-gestation, implantation sites in n-NOD.Ifng(-/-) and n-NOD mice are histologically similar. However, in d-NOD.Ifng(-/-), there is minimal development of spiral arteries, hypertrophy of the myometrial region containing uterine Natural Killer (uNK) cells and a deficit in cytoplasmic granule formation in the uNK cells. CONCLUSIONS Ifng contributes to the success of fertilization and to the rate of preimplantation mouse embryo development in normogylcemic and hyperglycemic pregnancies. A physiological role for this cytokine in human preimplantation development merits investigation.

4: The Journal of infectious diseases, 2010 Sep 2, 34(7)
Interferon gamma Responses to Mycobacterial Antigens Protect against Subsequent HIV-Associated Tuberculosis.

[Abstract]Background. The cellular immune responses that protect against tuberculosis have not been identified. Methods. We assessed baseline interferon gamma (IFN-gamma) and lymphocyte proliferation assay (LPA) responses to antigen 85 (Ag85), early secretory antigenic target 6 (ESAT-6), and Mycobacterium tuberculosis whole cell lysate (WCL) in human immunodeficiency virus (HIV)-infected and bacille Calmette-Gu��rin (BCG)-immunized adults with CD4 cell counts of 200 cells/muL who received placebo in the DarDar tuberculosis vaccine trial in Tanzania. Subjects were followed prospectively to diagnose definite or probable tuberculosis. Results. Tuberculosis was diagnosed in 92 of 979 subjects during a mean follow-up of 3.2 years. The relative risk of tuberculosis among subjects with positive IFN-gamma responses to Ag85 was 0.51 (95% confidence interval [CI], 0.26-0.99; [Formula: see text]), to ESAT-6 was 0.44 (95% CI, 0.23-0.85; [Formula: see text]), and to WCL was 0.67 (95% CI, 0.49-0.88; [Formula: see text]). The relative risk of tuberculosis was not significantly associated with baseline LPA responses. In a multivariate Cox regression model, subjects with IFN-gamma responses to ESAT-6 and WCL had a lower hazard of developing tuberculosis, with a hazard ratio for ESAT-6 of 0.35 (95% CI, 0.16-0.77; [Formula: see text]) and a hazard ratio for WCL of 0.30 (95% CI, 0.16-0.56; [Formula: see text]). Conclusions. Baseline IFN-gamma responses to ESAT-6 and WCL were associated with protection from subsequent tuberculosis among HIV-infected subjects with childhood BCG immunization in a region of high tuberculosis prevalence. Trial registration. ClinicalTrials.gov identifier: NCT00052195 .

5: Fitoterapia, 2010 Aug 18, 8(2)
Sesquiterpene lactone trilobolide activates production of interferon-gamma and nitric oxide.

[Abstract]Trilobolide (TB), a sesquiterpene lactone isolated from Laser trilobum is inhibitor of sarco/endoplasmic reticulum Ca2+-ATPase (SERCA). We revealed that upon the in vitro exposure to TB, rodent peritoneal cells and human peripheral blood mononuclear cells secreted high amounts of IFN-gamma The effect was associated with stimulation of high output NO biosynthesis in rat cells. The stimulatory potential of TB depends on activation of MAP kinases p38 and ERK1/2, and transcription factor NF-kappaB. BAPTA-AM, a chelator of the intracellular calcium, remained without any effect on secretion of IFN-gamma triggered by TB. These results demonstrate that TB is a potent immunostimulatory agent.

6: Cell host & microbe, 2010 Aug 19, 8(2)
Peptidoglycan Recognition Proteins Protect Mice from Experimental Colitis by Promoting Normal Gut Flora and Preventing Induction of Interferon-gamma.

[Abstract]There are multiple mechanisms that protect the intestine from an excessive inflammatory response to intestinal microorganisms. We report here that all four mammalian peptidoglycan recognition proteins (PGRPs or Pglyrps) protect the host from colitis induced by dextran sulfate sodium (DSS). Pglyrp1(-/-), Pglyrp2(-/-), Pglyrp3(-/-), and Pglyrp4(-/-) mice are all more sensitive than wild-type mice to DSS-induced colitis due to a more inflammatory gut microflora, higher production of interferon-gamma, higher expression of interferon-inducible genes, and an increased number of NK cells in the colon upon initial exposure to DSS, which leads to severe hyperplasia of the lamina propria, loss of epithelial cells, and ulceration in the colon. Thus, during experimental colitis, PGRPs protect the colon of wild-type mice from an early inflammatory response and the loss of the barrier function of intestinal epithelium by promoting normal bacterial flora and by preventing damaging production of interferon-gamma by NK cells in response to injury.

7: The Journal of biological chemistry, 2010 Aug 4, 73(1)
Interferon-gamma-mediated inhibition of serum response factor-dependent smooth muscle specific gene expression.

[Abstract]Interferon-gamma (IFNgamma) exerts multiple biological effects on effector cells by regulating many downstream genes, including smooth muscle specific genes. However, the molecular mechanisms underlying IFNgamma-induced inhibition of smooth muscle specific gene expression remain unclear. In this study, we have shown that serum response factor (SRF), a common transcriptional factor for cell proliferation, migration and differentiation is targeted by IFNgamma in a STAT1-dependent manner. We show that the molecular mechanism by which IFNgamma regulates SRF is via activation of the 2-5A-RNaseL system, which triggers SRF mRNA decay and reduced SRF expression. As a result, decreased SRF expression reduces expression of SRF target genes such as smooth muscle alpha-actin and smooth muscle myosin heavy chain. Additionally, IFNgamma reduced p300 and acetylated histone-3 binding in both smooth muscle alpha-actin and SRF promoters, epigenetically decreasing smooth muscle alpha-actin and SRF transcriptional activation. Our data reveal that SRF is a novel IFNgamma-regulated gene and further elucidate the molecular pathway between IFNgamma, IFNgamma-regulated genes, and SRF and its target genes.

8: PloS one, 2010, 5(7)
Signal Transducers and Activators of Transcription-1 (STAT1) Regulates microRNA Transcription in Interferon gamma-Stimulated HeLa Cells.

[Abstract]BACKGROUND: Constructing and modeling the gene regulatory network is one of the central themes of systems biology. With the growing understanding of the mechanism of microRNA biogenesis and its biological function, establishing a microRNA-mediated gene regulatory network is not only desirable but also achievable. METHODOLOGY: In this study, we propose a bioinformatics strategy to construct the microRNA-mediated regulatory network using genome-wide binding patterns of transcription factor(s) and RNA polymerase II (RPol II), derived using chromatin immunoprecipitation following next generation sequencing (ChIP-seq) technology. Our strategy includes three key steps, identification of transcription start sites and promoter regions of primary microRNA transcripts using RPol II binding patterns, selection of cooperating transcription factors that collaboratively function with the transcription factors targeted by ChIP-seq assay, and construction of the network that contains regulatory cascades of both transcription factors and microRNAs. PRINCIPAL FINDINGS: Using CAMDA (Critical Assessment of Massive Data Analysis) 2009 data set that includes ChIP-seq data on RPol II and STAT1 (signal transducers and activators of transcription 1) in HeLa S3 cells in control condition and with interferon gamma stimulation, we first identified promoter regions of 83 microRNAs in HeLa cells. We then identified two potential STAT1 collaborating factors, AP-1 and C/EBP (CCAAT enhancer-binding proteins), and further established eight feedback network elements that may regulate cellular response during interferon gamma stimulation. CONCLUSIONS: This study offers a bioinformatics strategy to provide testable hypotheses on the mechanisms of microRNA-mediated transcriptional regulation, based upon genome-wide protein-DNA interaction data derived from ChIP-seq experiments.

9: The journal of obstetrics and gynaecology research, 2010 Aug, 36(4)
Genetic contribution of the interferon gamma dinucleotide-repeat polymorphism in South Indian women with endometriosis.

[Abstract]Aim: To investigate whether the interferon-gamma (IFNG) gene dinucleotide (CA)-repeat polymorphism is responsible in part for genetic susceptibility to endometriosis in South Indian women. Methods: Following extraction of genomic DNA, genotyping of interferon-gamma CA-repeat polymorphism was performed using genescan technology. Results: The global IFNG allele frequencies in all patients with endometriosis were significantly different from those in the control women (chi(2) = 37.062; 6 degrees of freedom; P 10: Microbiology (Reading, England), 2010 Jul 23, 33(1)
Lactobacillus rhamnosus GG attenuates interferon-{gamma} and tumor necrosis factor-{alpha}-induced barrier dysfunction and pro-inflammatory signalling.

[Abstract]The intestinal epithelium forms a protective barrier against luminal contents and the external environment, mediated via intercellular tight junctions (TJ). The TJ can be disrupted from cell signalling induced by either enteric pathogens or pro-inflammatory cytokines, thereby contributing to various intestinal disorders ranging from acute infectious diarrhoea to chronic inflammatory bowel diseases. Probiotics, such as Lactobacillus rhamnosus GG (LGG), are reported to confer beneficial effects on epithelial cells including antagonizing infections and reducing overt pro-inflammatory responses, but the underlying mechanisms of these observed effects require further characterization. We hypothesized that probiotics preserve barrier function by interfering with pro-inflammatory cytokine signalling. Caco-2bbe cells were seeded into Transwells to attain polarized monolayers with intercellular TJ. Monolayers were inoculated apically with probiotic Lactobacillus rhamnosus GG (LGG) 3 h prior to the addition of IFN-gamma (100 ng ml-1) to the basolateral medium overnight. The monolayers were then placed in fresh basal medium +/- TNF-alpha (10 ng ml-1) and transepithelial electrical resistance (TER) measurements taken over the time course of TNF-alpha stimulation. To complement TER findings, cells were processed for zona occludens-1 (ZO-1) immunofluorescence staining. As a measure of TNF-alpha downstream signalling, cells were immunofluorescently stained for NF-kappaB p65 subunit and quantitation of CXCL-8 mRNA performed by qRT-PCR. Basal cell culture medium was collected after overnight TNF-alpha stimulation to measure secreted chemokines, including CXCL-8 (interleukin-8) and CCL-11 (eotaxin). Following LGG inoculation, IFN-gamma priming, and 24 h TNF-alpha stimulation, epithelial cells maintained TER and ZO-1 distribution. LGG diminished the nuclear translocation of p65, demonstrated by both immunofluorescence and CXCL-8 mRNA expression. CXCL-8 and CCL-11 protein levels were decreased in LGG-inoculated, cytokine-challenged cells. These findings indicate that LGG ameliorates the effects of pro-inflammatory cytokines on epithelial barrier integrity and inflammation mediated, at least in part, through inhibition of NF-kappaB signalling.

11: Thrombosis research, 2010 Jul 21, 33(1)
Interferon gamma in the etiology of atherosclerosis and periodontitis.

[Abstract]Clinical observations and a few research reports seem to suggest that intraoral infection as well as periodontal teeth could potentially lead to systemic infections including atherosclerosis. The aim of our investigations was to determine whether periodontal disease might aggravate atherosclerosis and whether interferon-gamma (IFNG), widely recognized as a potent multifunctional cytokine, might serve as a marker of the process. This is the first research based on tissue material such as atheromata and periodontal pocket granulation tissue. The study population consisted of 15 patients with periodontitis and atherosclerosis. Control group comprised 15 non-atherosclerotic patients with periodontitis. IFNG, IFNGR1 and IFNGR2 expression was analysed using qRT-PCR profiling in the inflammatory granulation tissue and atheroma. Granulation tissue samples obtained from non-atherosclerotic group showed a significant increase in IFNG and a decrease of IFNGR1, IFNGR2 expression whereas granulation tissue and atheromata of patients with systemic disease demonstrated lower IFNG and higher IFNGR1 and IFNGR2 expression.

12: Applied biochemistry and biotechnology, 2010 Jul 22, 33(1)
Development of DNA-Designed Avian IgY Antibodies for Quantitative Determination of Bovine Interferon-Gamma.

[Abstract]Interferon-gamma (IFN-gamma), a cytokine produced by sensitized T lymphocytes, is one of the key elements in defining T helper 1 lymphocyte immune responses. Quantitative evaluation of IFN-gamma expression could provide an important analytical tool for measurement of cell-mediated immunity and investigating immune responses to infectious diseases. Method of DNA-designed avian IgY antibodies was used for production of monospecific polyclonal antibodies that allows quantification of the recombinant bovine IFN-gamma protein. IFN-gamma cDNA was subcloned and expressed in mammalian expression plasmid (pcDNA3.1(+)) under the control of the human cytomegalovirus promoter. Chickens were immunized by plasmid DNA, and eggyolk antibodies extracted from eggs were collected after immunization. IgY-specific antibodies were evaluated by an antigen capture enzyme-linked immunosorbent assay (ELISA) using recombinant IFN-gamma. Based on the results, developed bovine IFN-gamma capture ELISA could detect up to 1 ng/ml of IFN-gamma by 64-fold diluted IgY. Monospecific anti-bovine IFN-gamma antibodies generated in chickens are useful for quantifying different concentrations of recombinant bovine IFN-gamma, which is expressed in cell culture.

13: Leukemia research, 2010 Jul 20, 33(1)
High fever and shock induced by interferon-gamma and interleukin-6 produced by adult T-cell leukaemia/lymphoma cells.

[Abstract]Interferon-gamma (IFN-gamma), a cytokine produced by sensitized T lymphocytes, is one of the key elements in defining T helper 1 lymphocyte immune responses. Quantitative evaluation of IFN-gamma expression could provide an important analytical tool for measurement of cell-mediated immunity and investigating immune responses to infectious diseases. Method of DNA-designed avian IgY antibodies was used for production of monospecific polyclonal antibodies that allows quantification of the recombinant bovine IFN-gamma protein. IFN-gamma cDNA was subcloned and expressed in mammalian expression plasmid (pcDNA3.1(+)) under the control of the human cytomegalovirus promoter. Chickens were immunized by plasmid DNA, and eggyolk antibodies extracted from eggs were collected after immunization. IgY-specific antibodies were evaluated by an antigen capture enzyme-linked immunosorbent assay (ELISA) using recombinant IFN-gamma. Based on the results, developed bovine IFN-gamma capture ELISA could detect up to 1 ng/ml of IFN-gamma by 64-fold diluted IgY. Monospecific anti-bovine IFN-gamma antibodies generated in chickens are useful for quantifying different concentrations of recombinant bovine IFN-gamma, which is expressed in cell culture.

14: Immunobiology, 2010 September -, 215(9-10)
Divergent modulation of Chlamydia pneumoniae infection cycle in human monocytic and endothelial cells by iron, tryptophan availability and interferon gamma.

[Abstract]Chlamydia pneumoniae is an obligatory intracellular bacterium causing chronic inflammatory diseases in humans. We studied the role of the nutritive factors, iron and tryptophan, towards the course of infection and immune response pathways in C. pneumoniae infected endothelial cells and monocytes. Human endothelial (EA.hy923) and monocytic cells (THP-1) were infected with C. pneumoniae, supplemented with iron or 1-methyltryptophan (1-MT), an inhibitor of the tryptophan degrading enzyme indoleamine 2,3-dioxygenase (IDO), and subsequently stimulated with IFN-gamma or left untreated. The number of infected cells, the morphology and quantity of C. pneumoniae inclusion bodies, IDO activity and innate immune effector pathways were analysed. While neither iron challenge, IDO inhibition or IFN-gamma treatment had a significant effect on C. pneumoniae morphology or numbers within THP-1 monocytic cells, iron supplementation to EA.hy926 cells resulted in promotion of C. pneumoniae proliferation and differentiation while IFN-gamma had an inhibitory effect. Furthermore, the number of infected endothelial cells was significantly decreased upon 1-MT treatment. C. pneumoniae infection induced a pro-inflammatory immune response as evidenced by increased IDO activity, neopterin formation or TNF-alpha production in THP-1 but not in endothelial cells. These pathways were superinduced upon IFN-gamma treatment and partly modulated by iron supplementation. Our results demonstrate that the infectious cycle of C. pneumoniae behaves differently between monocytic and endothelial cells. While the intracellular pathogen remains in a persistent form within monocytes, it can differentiate and proliferate within endothelial cells indicating that endothelial cells are a preferred environment for Chlamydia. Nutritive factors such as iron have subtle effects on C. pneumoniae biology in endothelial, but not monocytic cells. Our results contribute to a better understanding of C. pneumoniae infection and its role in chronic inflammatory diseases such as atherosclerosis.

15: Colloids and surfaces. B, Biointerfaces, 2010 Oct 15, 80(2)
Detecting interferon-gamma release from human CD4 T-cells using surface plasmon resonance.

[Abstract]Cytokine secretion by leukocytes is an important indicator of immune response to pathogens and therefore has significant implications in disease diagnostics. Given heterogeneity of leukocyte subsets and the ability of multiple cell subsets to secrete the same cytokines, connecting cytokine production to a specific leukocyte subset is a distinct challenge. In the present paper we describe a strategy combining antibody (Ab)-based affinity cell separation and surface plasmon resonance (SPR) for capturing human CD4 T-cells and for label-free detection of cell-secreted interferon (IFN)-gamma - an important inflammatory cytokine. Human blood was introduced into a flow chamber modified with anti-CD4 Abs resulting in capture of CD4(+) T-cells. After mitogenic activation of cells inside the flow chamber, culture medium was routed onto an SPR chip modified with monoclonal IFN-gamma Abs. SPR signal observed in this experiment correlated with cytokine production by T-cells. The strategy of combining SPR detection with cell purification may be used in the future for label-free, sensitive detection of multiple cytokines or proteins secreted by the desired cell subset.

16: The Journal of reproduction and development, 2010 Jun 16, 358(1-2)
Effects of Tumor Necrosis Factor alpha and Interferon gamma on the Viability and mRNA Expression of TNF Receptor Type I in Endothelial Cells from the Bovine Corpus Luteum.

[Abstract]The corpus luteum (CL) is mainly composed of luteal steroidogenic cells (LSCs) and luteal endothelial cells (LECs). Cell death of LSCs and LECs is essential for structural luteolysis. Therefore, it is important to understand the mechanisms regulating cell death in both types of luteal cells. We previously reported that a treatment combining tumor necrosis factor alpha (TNF) and interferon gamma (IFNG) induced cell death in LSCs and that LECs express TNF receptor type I (TNFRI). To investigate the mechanism of cell death in LECs, in the present study we determined the effects of the same cytokines on cell viability and TNFRI mRNA expression in cultured LECs. To induce cell death in LECs, LECs were treated with TNF or IFNG (0, 0.05, 0.5, 1.0, 2.5 nM) and a combination of TNF (0.5 nM) and IFNG (0.5 nM) for 24 h. The viability of LECs was reduced by a single treatment with TNF or IFNG dose-dependently (P<0.05). Cell viability was further decreased by treatment with a combination of TNF and IFNG (P<0.05). Cells with DNA fragmentation (TUNEL-positive cells) were observed after the treatment with TNF and IFNG. Semi-quantitative RT-PCR analysis revealed that treatment with IFNG alone or in combination with TNF increased the expression of TNFRI mRNA compared with the control (P<0.001). In summary, TNF and IFNG increased cell death in cultured bovine LECs. The upregulation of TNFRI mRNA expression by IFNG suggests that TNF and IFNG synergistically affect the death of LECs resulting in acute luteolysis.

17: Journal of neuroimmunology, 2010 Aug 25, 225(1-2)
Differential expression of interferon-gamma receptor on human glial cells in vivo and in vitro.

[Abstract]Although significant effects of interferon-gamma (IFN-gamma) on glial cells are well documented, information on the expression level and localization of glial IFN-gamma receptors (IFN-gamma-R) in the human central nervous system (CNS) is sparse. To examine the glial expression of IFN-gamma-R in the human CNS, immunohistochemistry and quantitative analyses were performed on Alzheimer disease hippocampus, Parkinson disease substantia nigra, amyotrophic lateral sclerosis spinal cord and corresponding areas from non-neurological cases. Almost all IFN-gamma-R-positive (IFN-gamma-R(+)) cells corresponded to GFAP-positive (GFAP(+)) astrocytes, while none of IFN-gamma-R(+) cells corresponded to IBA1-positive (IBA1(+)) microglia or MBP-positive (MBP(+)) oligodendrocytes in these neurological cases. We observed a similar pattern of glial IFN-gamma-R expression in non-neurological cases. Also, we quantitatively analyzed the IFN-gamma-R expression by cultured human glial cells using immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). In contrast to in vivo results, almost all IFN-gamma-R(+) cells were IBA1(+) in microglial cultures, GFAP(+) in astrocytic cultures and MBP(+) in oligodendrocytic cultures. Moreover, no significant difference in IFN-gamma-R mRNA expression was found for these glial cell types by RT-PCR. These results suggest that the microglial and oligodendrocytic expression levels of IFN-gamma-R are much lower than the astrocytic expression levels in the human CNS in vivo, whereas all three types of glial cells constitutively express IFN-gamma-R when cultured in vitro.

18: Clinical journal of the American Society of Nephrology : CJASN, 2010 Jun 10, 358(1-2)
High Prevalence of Latent Tuberculosis Infection in Dialysis Patients Using the Interferon-{gamma} Release Assay and Tuberculin Skin Test.

[Abstract]BACKGROUND AND OBJECTIVES: Patients in ESRD on hemodialysis with latent tuberculosis (TB) infection have 10 to 25 times the risk of reactivation into active disease compared with healthy adults. This study investigates the prevalence of latent TB infection in dialysis patients from a country with an intermediate burden of TB and its associated risk factors using the QuantiFERON-TB Gold in-tube test (QGIT) and the tuberculin skin test (TST). DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: This was a prospective, cross-sectional study performed at a medical center in Taiwan on dialysis patients. Each patient underwent QGIT, two-step TST using 2 tuberculin units (TU) of PPD RT-23, a chest x-ray to exclude active TB, and an interview to determine TB risk factors. RESULTS: Ninety-three of 190 eligible patients were enrolled: 35 men and 58 women. 64.8% were vaccinated with the Bacille-Calmette-Gu��rin (BCG) vaccination. Overall, 34.4% were positive by QGIT and 10.8% were indeterminate. Using a 10-mm TST cutoff, 53.9% were positive. There was poor correlation between TST and QGIT at any TST cutoff criteria. There was a significant increasing trend of QGIT positivity with age in those younger than 70 years, and, conversely, a decreasing trend of TST reactivity with age. Significant risk factors for QGIT positivity included age and past TB disease. CONCLUSIONS: This study shows a high prevalence of latent TB infection in dialysis patients in a country with an intermediate burden of TB. QGIT in dialysis patients correlated better than TST with the risk of TB infection and past TB disease.

19: Archives of gynecology and obstetrics, 2010 May 27, 71(6)
Dose-response effect of interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha, and interferon-gamma on the in vitro production of epithelial neutrophil activating peptide-78 (ENA-78), IL-8, and IL-6 by human endometrial stromal cells.

[Abstract]PURPOSE: The production of epithelial neutrophil activating peptide-78 (NA-78) and the interleukins IL-8 and IL-6 by endometrial stromal cells is stimulated by pro-inflammatory interleukin-1 (IL-1) and tumour necrosis factor-alpha (TNF-alpha). IL-8 is suggested to play a role in the pathogenesis of endometriosis, and in these women the peritoneal fluid concentrations of ENA-78 and IL-8 are increased. TNF-alpha has been tested together with interferon-gamma because of their cooperative stimulation of IL-6. The release of IL-8, however, is inhibited with increasing interferon levels. The aim of the study was the analysis of the production of ENA-78, IL-6 and IL-8 by cultured human endometrial stromal cells in the presence of varying concentrations of IL-1beta, TNF-alpha, and interferon-gamma. METHODS: Eutopic endometrial tissue was obtained from seven cycling, endometriosis-free women undergoing laparoscopy for reasons of infertility or pain. The release of ENA-78, IL-8 and IL-6 by the isolated and monolayer cultured stromal cell fraction in the presence of IL-1beta (0.08 to 50 ng/mL), TNF-alpha, and interferon-gamma (both 20 to 500 ng/mL) was determined. RESULTS: IL-1beta stimulated the production of IL-8, IL-6, and ENA-78 dose dependently from 0.08 to 2.0 ng/mL (ENA-78) or to 10 ng/mL (IL-8, IL-6); at 50 ng/mL a decrease in release was observed for IL-8 and IL-6. TNF-alpha stimulation yielded a plateau between 20 and 100 ng/mL. Interferon-gamma stimulated IL-6 and inhibited IL-8 production above 20 ng/mL. ENA-78 release was largely unaffected by interferon-gamma. CONCLUSIONS: IL-1beta and TNF-alpha stimulate stromal cytokine production cumulatively with different dose-response curves. The presence of interferon-gamma has opposite effects on IL-8 and IL-6. TNF-alpha and interferon-gamma should be investigated separately in future in vitro studies with endometrial cells and explants.

20: Molecular biology of the cell, 2010 May 26, 71(6)
The Interferon-{gamma}-induced Murine Guanylate-Binding Protein-2 (mGBP-2) Inhibits Rac Activation during Cell Spreading on Fibronectin and Platelet-derived Growth Factor (PDGF) Treatment: Role for Phosphatidylinositol 3-Kinase (PI3-K).

[Abstract]Monitoring Editor: Josephine C. Adams Exposure of cells to certain cytokines can alter how these same cells respond to later cues from other agents, such as ECM or growth factors. Interferon-gamma pre-exposure inhibits the spreading of fibroblasts on fibronectin. Expression of the IFN-gamma-induced GTPase, mGBP-2, can phenocopy this inhibition and siRNA knockdown of mGBP-2 prevents IFN-gamma-mediated inhibition of cell spreading. Either IFN-gamma treatment or mGBP-2 expression inhibits Rac activation during cell spreading. Rac is required for cell spreading. mGBP-2 also inhibits the activation of Akt during cell spreading on fibronectin. mGBP-2 is incorporated into a protein complex containing the catalytic subunit of PI3-K, p110. The association of mGBP-2 with p110 appears important for the inhibition of cell spreading because S52N mGBP-2, which does not incorporate into the protein complex with p110, is unable to inhibit cell spreading. PI-3K activation during cell spreading on fibronectin was inhibited in the presence of mGBP-2. Both IFN-gamma and mGBP-2 also inhibit cell spreading initiated by PDGF treatment, which is also accompanied by inhibition of Rac activation by mGBP-2. This is the first report of a novel mechanism by which IFN-gamma can alter how cells respond to subsequent extracellular signals, by the induction of mGBP-2.

21: The Journal of infectious diseases, 2010 May 26, 71(6)
Interferon gamma and Interleukin 4 Have Contrasting Effects on Immunopathology and the Development of Protective Adaptive Immunity against Mycoplasma Respiratory Disease.

[Abstract]For vaccine development, it is critical to understand the regulatory mechanisms determining resistance and immunopathology against mycoplasma respiratory diseases. The present study evaluated the contribution of the polarizing cytokines interferon gamma (IFN-gamma) and interleukin 4 (IL-4) in the regulation of mycoplasma-specific immunity. The absence of a single cytokine (either IFN-gamma or IL-4) uniquely altered the expression of multiple chemokines and cytokines in the lungs of uninfected mice and influenced responses to mycoplasma infection. Most importantly, prior nasal-pulmonary immunization of IFN-gamma(-/-) mice led to exacerbated mycoplasma disease, whereas immunized IL-4(-/-) mice were dramatically more resistant than wild-type mice. Helper T cell type 2 responses in IFN-gamma(-/-) mice corresponded to immunopathologic reactions that developed after mycoplasma infection or immunization. Thus, adaptive immunity clearly can independently promote either protection or immunopathology against mycoplasma infection, and optimal vaccination appears to be dependent on promoting protective IFN-gamma-dependent networks (perhaps helper T cell type 1 responses) while minimizing the effect of IL-4-mediated responses, which dampen the generation of protective immunity.

22: The American journal of pathology, 2010 May 20, 14(6)
N-Acetylcysteine and 15 Deoxy-{Delta}12,14-Prostaglandin J2 Exert a Protective Effect Against Autoimmune Thyroid Destruction In Vivo but Not Against Interleukin-1{alpha}/Interferon {gamma}-Induced Inhibitory Effects in Thyrocytes In Vitro.

[Abstract]Reactive oxygen species (ROS) are crucial for thyroid hormonogenesis, and their production is kept under tight control. Oxidative stress (OS) is toxic for thyrocytes in an inflammatory context. In vitro, Th1 pro-inflammatory cytokines have already been shown to decrease thyroid-specific protein expression. In the present study, OS level and its impact on thyroid function were analyzed in vitro in Th1 cytokine (interleukin [IL]-1alpha/interferon [IFN] gamma)-incubated thyrocytes (rat and human), as well as in vivo in thyroids from nonobese diabetic mice, a model of spontaneous autoimmune thyroiditis. N-acetylcysteine (NAC) and prostaglandin, 15 deoxy-(Delta12,14)-prostaglandinJ2 (15dPGJ2), were used for their antioxidant and anti-inflammatory properties, respectively. ROS production and OS were increased in IL-1alpha/IFNgamma-incubated thyrocytes and in destructive thyroiditis. In vitro, NAC not only reduced ROS production below control levels, but further decreased the expression of thyroid-specific proteins in addition to IL-1alpha/IFNgamma-inhibitory effects. Thus, besides ROS, other intracellular intermediaries likely mediate Th1 cytokine effects. In vivo, NAC and 15dPGJ2 reduced OS and the immune infiltration, thereby leading to a restoration of thyroid morphology. It is therefore likely that NAC and 15dPGJ2 mainly exert their protective effects by acting on infiltrating inflammatory cells rather than directly on thyrocytes.

23: Current opinion in pulmonary medicine, 2010 May 13, 135(1-2)
Use of pleural fluid levels of adenosine deaminase and interferon gamma in the diagnosis of tuberculous pleuritis.

[Abstract]PURPOSE OF REVIEW: This review aims to define the role of adenosine deaminase (ADA) and interferon gamma (IFN-gamma) in the differential diagnosis of pleural effusion with special attention to their source, mechanism of release and methods of measurement in pleural fluid. The diagnostic performance of ADA and IFN-gamma is analyzed, and the advantages and limitations of their use in differentiating between tuberculous and nontuberculous pleural effusion are disucussed. RECENT FINDINGS: Several potential biomarkers of tuberculous pleurisy have been evaluated, but none have been found to be clearly superior to pleural fluid level of ADA or IFN-gamma. The majority of recent studies confirm the high diagnostic utility of pleural fluid ADA and IFN-gamma measurement; hence, these markers are included in different diagnostic algorithms for patients suspected of tuberculous pleurisy. Other relatively new tests show a high variability [nucleic acid amplification tests (NAATs)] or are technically demanding, costly and give equivocal results in patients with active tuberculosis [IFN-gamma releasing assays (IGRAs)]. SUMMARY: Pleural fluid ADA and IFN-gamma are both sensitive and specific biomarkers of tuberculous pleurisy. Their diagnostic accuracy across the different studies shows a smaller variability than that of other tests, for example NAATs. There is also no convincing evidence that IGRAs are superior to pleural fluid ADA or IFN-gamma measurement. Hence, the role of ADA and IFN-gamma in the differential diagnosis of tuberculous pleurisy is pivotal.

24: Blood, 2010 May 14, 135(1-2)
Interferon-{gamma} and tumor necrosis factor-{alpha} induce an immunoinhibitory molecule, B7-H1, via nuclear factor {kappa}B activation in blasts in myelodysplastic syndromes.

[Abstract]During disease progression in myelodysplastic syndromes (MDS), clonal blasts gain a more aggressive nature, while nonclonal immune cells become less efficient, via an unknown mechanism. Using MDS cell lines and patient samples, we showed that the expression of an immunoinhibitory molecule, B7-H1 (CD274), was induced by interferon-gamma (IFNgamma) and tumor necrosis factor-alpha (TNFalpha) on MDS blasts. This induction was associated with the activation of nuclear factor kappaB (NF-kappaB) and nearly completely blocked by an NF-kappaB inhibitor, pyrrolidine dithiocarbamate. B7-H1(+) MDS blasts had greater intrinsic proliferative capacity than B7-H1(-) MDS blasts when examined in various assays. Furthermore, B7-H1(+) blasts suppressed T-cell proliferation and induced T-cell apoptosis in allogeneic cocultures. When fresh bone marrow samples from patients were examined, blasts from high-risk MDS patients expressed B7-H1 molecules more often compared with those from low-risk MDS patients. Moreover, MDS T cells often overexpressed PD-1 molecules that transmit an inhibitory signal from B7-H1 molecules. Taken together, these findings provide new insight into MDS pathophysiology. IFNgamma and TNFalpha activate NF-kappaB that in turn induces B7-H1 expression on MDS blasts. B7-H1(+) MDS blasts have an intrinsic proliferative advantage and induce T-cell suppression, which may be associated with disease progression in MDS.

25: Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 2010 Mar 29, 30(6)
ROLE OF THE IFNG +874T/A POLYMORPHISM IN CHAGAS DISEASE IN A COLOMBIAN POPULATION.

[Abstract]Genetic susceptibility to Trypanosoma cruzi infection and the development of cardiomyopathy is complex, heterogeneous, and likely involves several genes. Previous studies have implicated cytokine and chemokine genes in susceptibility to Chagas disease. Here we investigated the association between the interferon-gamma gene (IFNG) +874T/A polymorphism and Chagas disease, focusing on susceptibility and severity. This study included 236 chagasic patients (asymptomatic, n=116; cardiomyopathic, n=120) and 282 healthy controls from a Colombian population where T. cruzi is highly endemic. Individuals were genotyped for functional single nucleotide polymorphism (SNP; rs2430561; A/T) of the IFNG gene by amplification refractory mutational system PCR (ARMS-PCR). Moreover, clinical manifestations of Chagas in patients were analysed. We found a significant difference in the distribution of the IFNG +874 "A" allele between patients and healthy controls (P=0.003; OR=1.46, 95% CI, 1.13-1.89). The frequency of the IFNG +874 genotype A/A, which is associated with reduced production of interferon-gamma, was increased in the patients relative to controls (38.1% vs. 26.6%). We compared the frequencies of IFNG alleles and genotypes between asymptomatic patients and those with chagasic cardiomyopathy and found no significant difference. Our data suggest that the IFNG +874T/A genetic polymorphism may be involved in susceptibility but not in the progression of Chagas disease in this Colombian population.

26: Molecular diagnosis & therapy, 2010 Apr 1, 14(2)
Chronic Ulceration from Mycobacterium marinum Infection and the Diagnostic Value of T-Cell Interferon-gamma Release Assays.

[Abstract]This case report describes the differential diagnosis of cutaneous ulcerations and the utility of the interferon-gamma release assays as a tool to aid in the diagnosis. These new assays can be used to identify mycobacterial infections (specifically Mycobacterium marinum) as the etiologic agents.

27: Veterinary immunology and immunopathology, 2010 Jul, 136(1-2)
Bovine tuberculosis: Effect of the tuberculin skin test on in vitro interferon gamma responses.

[Abstract]Bovine tuberculosis (bTB) is a disease of zoonotic and economic importance. In many countries, control is based on test and slaughter policies and/or abattoir surveillance. For testing, cell mediated immune- (CMI-) based assays (i.e., tuberculin skin test (TST) supplemented by the interferon gamma (IFN-gamma) assay) are the primary surveillance and disease control tests for bTB. The combined use of the in vivo and in vitro CMI assays to increase overall sensitivity has raised the question of whether the IFN-gamma response is influenced by injection of purified protein derivatives (PPDs) for TST. Published data on the influence of the TST, applied as the caudal fold test (CFT) or the comparative cervical test (CCT), on the IFN-gamma assay are contradictory. Reviewing published data and including additional data, the following conclusions can be drawn: (1) in naturally infected cattle, PPD administration for the single or repeated short-interval CCT neither boosts nor depresses PPD-specific IFN-gamma production. Disparate results have been concluded from some studies using experimental infections, emphasizing the importance of confirming initial experimental-based findings with studies using cattle naturally infected with Mycobacterium bovis. (2) In cattle experimentally infected with M. bovis, PPD administration for CFT boosts PPD-specific IFN-gamma production for up to 7 days without any effect on test interpretation. Importantly, in naturally infected cattle, CFT-related boosting selectively increases the in vitroM. bovis PPD (PPD-B) response 3 days after CFT, resulting in an increased PPD-B response relative to the response to Mycobacterium avium PPD (PPD-A). In non-infected cattle, it cannot be excluded that the CFT induces a mild boost of the PPD-specific response, particularly in animals sensitized to environmental, non-tuberculous mycobacteria, thus decreasing the specificity of the IFN-gamma assay. (3) In general, there is a lack of data clearly characterizing the effect of TSTs on the IFN-gamma assay. Further studies are required to clearly describe the effects of both CFT and CCT in non-infected animals and in naturally infected cattle, especially in low reacting infected cattle.

28: Veterinary immunology and immunopathology, 2010 Jul, 136(1-2)
Relative quantitative kinetics of interferon-gamma and interleukin-10 mRNA and protein production by activated ovine peripheral blood mononuclear cells.

[Abstract]Interferon-gamma (IFN-gamma) and interleukin (IL)-10 are cross-regulatory cytokines capable of driving and controlling the adaptive host immune response. The inter-relationship between IFN-gamma and IL-10 expression has not been defined in sheep despite biological evidence suggesting that they perform similar functions to their orthologues described in other species. To address this, we have developed a quantitative (q)PCR method to assess relative levels of IFN-gamma and IL-10 mRNA expression in activated ovine peripheral blood mononuclear cells (PBMC) and compared the kinetics of mRNA expression with amounts of cytokine secreted by the cells over a 96h period. PBMC were collected from sheep immunised with the nominal antigen ovalbumin (Ova) and re-stimulated in vitro with antigen and the T cell mitogen concanavalin A (ConA). The recall response to antigen was characterised by a single peak in IFN-gamma mRNA expression at 48h of culture (13-fold increase over unstimulated cells) and relatively lower expression of IL-10 mRNA (average 2-3-fold increase over the 96h culture period). Antigen-driven IFN-gamma protein concentration was greatest at the end of the culture period (96h) whereas IL-10 protein level was not elevated above that observed in unstimulated cells. The typical response to ConA was greater for both cytokines, with IFN-gamma mRNA expression peaking at 6h of culture (133-fold increase) then declining rapidly whereas IL-10 mRNA expression peaked at 24h (16-fold increase) and declined more gradually. Despite these differences in the relative kinetics of mRNA expression in mitogen-activated PBMC, the typical pattern of protein expression of the two cytokines was similar. Both showed a gradual rise in protein concentration starting from 12h of culture which was still rising at the end of the culture period (96h). These data demonstrate that the kinetics of mRNA expression for IFN-gamma and IL-10 in activated ovine PBMC do not necessarily correlate with detectable protein in culture.

29: Journal of immunological methods, 2010 Feb 26, 104(3)
Validation of a miniaturized assay based on IFNg secretion for assessment of specific T cell immunity.

[Abstract]A miniaturized method for detection of antigen induced secretion of IFNg by specific T cells cultured in 384 well plates has been recently reported. In order to confidently apply this assay to clinical investigations for monitoring of specific T cell immunity, an intralaboratory validation study has been undertaken. High reproducibility and linearity of reference curves was demonstrated. Consecutive replicate experiments handled by different operators using broad panels of recall antigens were reproducible when tested on individual biological samples. Kinetics of IFNg secretion with different antigens showed a plateau after 24 hour culture. Similar trends were observed with secretion of TNFa, GM-CSF and IL17, suggesting that the same kinetics can be applied if other cytokines are tested with this assay. It was demonstrated that frozen-thawed cells can be tested by cell-ELISA and that when PBMC are replaced by whole blood similar reactivity profiles were observed even though cytokine concentration was lower. T cell responses were higher in round bottom than in flat bottom wells, but these plates could not be applied to cell-ELISA as clear plates are not available for scanning. In conclusion, the assay proved flexible, since plates can be frozen at different times during the process, fresh or frozen PBMC and PBMC or whole blood could be used, and robust, since reproducibility was remarkable even when different operators performed the procedures.

30: American journal of reproductive immunology (New York, N.Y. : 1989), 2010 Feb 28, 104(3)
Interferon-gamma Inhibits Metalloproteinase Activity and Cytotrophoblast Cell Migration.

[Abstract]Citation Fontana VA, Sanchez M, Cebral E, Calvo JC. Interferon-gamma inhibits metalloproteinase activity and cytotrophoblast cell migration. Am J Reprod Immunol 2010 Problem Establishment of a successful pregnancy relies on a complex fetal-mother communication that starts with the embryo adhering and invading the endometrium. This requires remodeling of extracellular matrix, performed by metalloproteinases. Cytokines, such as interferon-gamma (IFN-gamma), play a role in implantation and could affect the success of pregnancy. Method of study Using JEG-3 cell line as model, we cultured the cells in the presence or absence of IFN-gamma and determined the activities of MMP-2 and MMP-9 using zymography and the secretion of leptin using Western blot. Results Interferon-gamma inhibits gelatinase activity from MMP-2 and MMP-9 in a dose-dependent manner, reducing the secretion of leptin (not because of a general inhibition on protein synthesis) and impairs cell migration on Matrigel. Conclusion Our results correlate with previous reports from our laboratory indicating that IFN- gamma is deleterious for mouse embryo outgrowth, having an effect on metalloproteinases activity as well as leptin secretion.

31: PloS one, 2010, 5(2)
Role of QuantiFERON-TB Gold, Interferon Gamma Inducible Protein-10 and Tuberculin Skin Test in Active Tuberculosis Diagnosis.

[Abstract]BACKGROUND: The measurement of Interferon gamma or Interferon gamma inducible protein (IP)-10 in antigen stimulated blood samples is suggested as an alternative method for latent tuberculosis (TB) diagnosis. Nonetheless, their role in active TB diagnosis, particularly in TB endemic settings is yet to be defined. In this study, the sensitivities and specificities of Interferon gamma release assay (IGRA), IP-10 assay and tuberculin skin test (TST) in detecting active TB cases were assessed in human immunodeficiency virus (HIV) sero-negative TB patients and healthy controls respectively. METHODS/PRINCIPAL FINDINGS: A total of 177 adult TB patients and 100 healthy controls were included for this study. QuantiFERON-TB Gold In-tube (QFT-IT) method was used to analyze the sensitivity and specificity of IGRA. QFT-IT, IP-10 and TST yielded the diagnostic sensitivities of 90.6% (95%CI: 86.3%-94.9%), 92.5% (95%CI: 88.6%-96.4%) and 68.9% (95%CI: 60.6%-77.2%) and specificities of 55% (95% CI: 35.2%-54.8%), 48% (95% CI: 38.2%-57.8%) and 75.5% (95% CI: 66.8%-84.2%), respectively. The extent of pulmonary involvement or presence of diabetes mellitus did not appear to influence the sensitivities of any of these tests. The combination of any of the two tests among QFT-IT, IP-10 and TST showed >98% sensitivity among smear negative cases and particularly the combination of IP-10, TST and smear microscopy showed 100% sensitivity, however, the specificity was decreased to 44.8%. CONCLUSIONS/SIGNIFICANCE: QFT-IT and IP-10 were highly sensitive in detecting active TB cases. The combination with TST improved the sensitivity of QFT-IT and IP-10 significantly. Although the higher sensitivity of combination of QFT-IT/IP-10 and TST may be useful in active TB diagnosis, they are limited by their poor specificity due to the high prevalence of latent TB in our settings.

32: Respiration; international review of thoracic diseases, 2010 Jan 21, 285(4)
HRCT and Whole-Blood Interferon-gamma Assay for the Rapid Diagnosis of Smear-Negative Pulmonary Tuberculosis.

[Abstract]Background: Early diagnosis of active pulmonary tuberculosis (PTB) is critical for TB control, and difficult in patients with smear-negative sputum. Objective: We wanted to evaluate the usefulness of clinical findings, high-resolution computed tomography (HRCT), interferon-gamma-releasing assay (IGRA) and polymerase chain reaction (PCR) of sputum in the diagnosis of smear-negative PTB. Methods: From June 2006 to September 2008, 178 patients with suspected PTB on the basis of clinical and radiological findings visited our institute. After excluding smear-positive cases (n = 77) and cases with an inconclusive diagnosis (n = 17), we studied 84 patients. Their clinical records, HRCT, sputum TB-PCR assay and IGRA results were retrospectively evaluated. A QuantiFeron-TB Gold (QFT-G; Cellestis Ltd., Carnegie, Vic., Australia) assay was used for the IGRA. Results: Active PTB was diagnosed in 40 (48%) of 84 patients; lack of sputum and young age were significantly associated with an increased risk of PTB. The sensitivities of sputum PCR assay, IGRA, and HRCT were 43.2, 84.4 and 80.0%, respectively, and the specificities were 97.7, 82.9 and 70.5%, respectively. Among the 38 patients suspected of having PTB based on HRCT, 24 patients showed positive results on the IGRA, and 23 of these were diagnosed with active PTB. Among the 35 patients suggested not to have TB based on HRCT, 25 showed negative results on the IGRA, and 23 (92%) of these were diagnosed as not to have TB. Conclusion: The combined results of HRCT and the IGRA could help decision-making for early initiation of treatment in smear-negative patients.

33: European journal of orthodontics, 2010 Jan 27, 22(1)
Interferon-{gamma}-loaded collagen scaffolds reduce myofibroblast numbers in rat palatal mucosa.

[Abstract]Wound contraction and scar formation after cleft palate repair lead to growth impairment of the maxilla and midface. Myofibroblasts play a key role in these processes. The application of an interferon-gamma (IFN-gamma)-loaded collagen scaffold after surgery might reduce the differentiation of myofibroblasts. In this study, the tissue response to IFN-gamma-loaded collagen scaffolds was evaluated after implantation in the palate of rats. Scaffolds, with or without IFN-gamma, were implanted submucoperiosteally in the palate of two groups of 25 five-week-old male Wistar rats. Groups of five rats were sacrificed at 1, 2, 4, 8, and 16 weeks post-implantation and processed for histological analyses. On haematoxylin and eosin-stained sections, the cell density and number of giant cells within the scaffolds were determined. Blood vessels, inflammatory cells, and myofibroblasts were detected by immunohistochemistry. The data for cell density, blood vessels, and giant cells were compared with a two-way analysis of variance. The scores for myofibroblasts and inflammation were compared by a rank sum test. A mild and rapidly subsiding inflammatory and foreign body response was found in both groups. Angiogenesis had already begun after 1 week, showed a peak after 4 weeks, and declined thereafter. IFN-gamma induced a faster influx of host cells and a major reduction in myofibroblast numbers. The scaffolds might be suitable for future applications in oral surgery.

34: The Journal of urology, 2010 Jan 21, 285(4)
Crucial Role of Interferon-gamma in Experimental Autoimmune Prostatitis.

[Abstract]PURPOSE: An autoimmune etiology is proposed in some patients with chronic nonbacterial prostatitis since they show interferon-gamma secreting lymphocytes specific to prostate antigens in the periphery and increased interferon-gamma in seminal plasma. We investigated the involvement of interferon-gamma in an animal model of autoimmune prostatitis. MATERIALS AND METHODS: Experimental autoimmune prostatitis was studied in the no-obese diabetic and C57Bl/6 (Harlan, Zeist, The Netherlands) susceptible mouse strains, and in the IRF-1 KO and STAT-1 KO mouse strains deficient in transcription factors involved in interferon-gamma signaling. RESULTS: Experimental autoimmune prostatitis was characterized by prostate specific interferon-gamma secreting cells in the periphery and by T-helper 1 related cytokines in the target organ. Increased interferon-gamma and interleukin-12 were observed in the prostate of autoimmune animals while interleukin-10 and interleukin-4 were decreased and unaltered, respectively. The absence of transcription factors involved in the interferon-gamma signaling cascade, IRF-1 and STAT-1, made mice resistant to experimental autoimmune prostatitis. IRF-1 KO and STAT-1 KO mice immunized with prostate antigens did not show infiltration or alterations in the prostate. They did not have the typical prostate specific autoimmune response and showed decreased interferon-gamma, interleukin-12 and interleukin-10, and augmented interleukin-4 in the prostate. CONCLUSIONS: Our results argue for a crucial role of interferon-gamma as a key factor in the pathogenesis of the disease. Intense research is promptly required to identify the pathogenic mechanisms underlying chronic prostatitis/chronic pelvic pain syndrome to find a more rational therapy.

35: The Tohoku journal of experimental medicine, 2010 Jan, 220(1)
High numbers of interferon-gamma-producing T cells and low titers of anti-tuberculous glycolipid antibody in individuals with latent tuberculosis.

[Abstract]Latent tuberculosis infection (LTBI) is defined as an infection with Mycobacterium tuberculosis (MTB) without clinical, bacteriological, or radiological findings, and its early diagnosis is essential for eradication of tuberculosis. To identify LTBI, we measured the numbers of interferon-gamma producing T cells, based on the ELISPOT assay, and the antibody titers in the sera to tuberculous glycolipid antigen (TBGL-Ab). Seventeen culture-confirmed TB patients, 13 controls from TB endemic areas (EC) and 13 controls from TB non-endemic areas (NEC) were enrolled. Peripheral blood mononuclear cells (2.5 x 10(5) per well) were cultured on plates precoated with antibody against interferon-gamma. ELISPOT response was defined as positive when the MTB-specific antigen-containing wells showed at least 6 spots and twice numbers of spots than negative control wells. ELISPOT responses were positive in 15 (88%), 8 (62%) and 4 (31%) subjects of TB, EC and NEC groups, respectively. The ELISPOT data differ between TB and NEC groups (p < 0.01) but not between TB and EC groups. In contrast, TBGL-Ab titers were elevated (> 2.0 U/ml) in 12 TB patients (71%), but only in one subject (8%) each from EC and NEC groups. These results indicate the high prevalence of LTBI in EC. In conclusion, LTBI is associated with positive ELISPOT assay and the low titer of TBGL-Ab, while positive results both in ELISPOT and TBGL-Ab assays indicate active TB. The low titer of TBGL-Ab is a helpful marker to identify LTBI in ELISPOT-positive individuals in TB endemic areas.

36: Gynecologic oncology, 2009 Dec 1, 58(12)
Interaction of immunological genes on chromosome 2q33 and IFNG in susceptibility to cervical cancer.

[Abstract]OBJECTIVE: Cervical cancer is caused by persistent infection with human papillomavirus and genetic susceptibility factors may augment disease risk. The immune response consists of complex interactions and it was recently proposed that the association of combinations of genotypes at several genes should be examined. In support of this the combination CD28+17(TT)/IFNG+874(AA) was shown to increase cervical cancer risk in a Brazilian population (VB Guzman et al. New approach reveals CD28 and IFNG gene interaction in the susceptibility to cervical cancer. Hum Mol Genet 2008;17:1838-44) and our aim was to replicate this finding. METHODS: We re-examined the proposed associations by analysis of polymorphisms at CD28, IFNG, TNF, PDCD1, ICOS and CTLA4 in 1306 Swedish cases and 811 controls. RESULTS: Logistic regression analysis detected association at single SNP level for CD28+17 (p=0.01), IFNG+874 (p=0.02), and PDCD1+7785 (p=0.04). The two locus combination CD28+17(TT)/IFNG+874(AA) (OR=0.76 (0.60-0.96, empirical p=0.03) and the three-locus combination CD28+17(TT)/IFNG+874(AA)/ICOS+1564(TT) (OR=0.65(0.49-0.87), empirical p=0.006) were associated with decreased risk. The strongest association was detected for the combination CTLA4-319 (CC)/IFNG (AA) (OR=0.67(0.53-0.84), empirical p=0.0007). CONCLUSION: The observation that these combinations of loci are associated in different populations supports their importance in cervical cancer development although the opposite directions of the effect call for clarification. The polymorphisms studied might not be the functional variants per se, but linked to those exerting a functional effect. The opposite associations in the two populations could then be explained by differences in linkage disequilibrium and population structure.

37: International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases, 2009 Nov 23, 57(22)
Treatment of relapsing Mycobacterium avium infection with interferon-gamma and interleukin-2 in an HIV-negative patient with low CD4 syndrome.

[Abstract]A patient with idiopathic CD4 T-lymphopenia was diagnosed with a recurrent disseminated Mycobacterium avium infection. Because of progressive disease, treatment with interferon-gamma (IFN-gamma) and interleukin-2 (IL-2) was started. The patient was successfully treated with IFN-gamma-1b and IL-2 in addition to anti-mycobacterial combination therapy. To our knowledge, this is the first report of successful combination therapy with IFN-gamma-1b and IL-2 in a patient with idiopathic CD4 T-lymphopenia. Short-term IFN-gamma-1b and IL-2 might be considered as therapeutic options in refractory mycobacterial infections in patients with idiopathic CD4 lymphopenia.

38: Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research, 2009 Nov 24, 57(22)
Roles of IKK-beta, IRF1, and p65 in the Activation of Chemokine Genes by Interferon-gamma.

[Abstract]Activation of chemokine genes in response to interferon (IFN)-gamma or NF-kappaB is an important aspect of inflammation. Using the chemokine gene ip-10 in mouse embryonic fibroblast cells as an example, we show that the response to IFN-gamma is long lasting but secondary: initial STAT1 activation drives IRF1 synthesis, and IRF1 then binds to IFN-stimulated regulatory elements (ISREs) in the ip-10 promoter. The promoters of most IKK-beta-dependent IFN-stimulated genes (ISGs) also contain ISREs. In response to IFN-gamma, inhibitor of NF-kappaB (IkappaB) kinase beta (IKK-beta) is required to activate both newly synthesized IRF1 and the p65 subunit of NF-kappaB, which contributes to ip-10 expression by binding to kappaB sites in the ip-10 promoter, with little or no activation of classical NF-kappaB. In contrast to IFN-gamma, IL-1beta induces ip-10 expression rapidly but transiently, by activating classical NF-kappaB and increasing the synthesis of IRF1. Together, IL-1beta and IFN-gamma induce ip-10 synergistically. IFN-gamma does not affect the transient activation of classical NF-kappaB by IL-1beta and synergistic induction of ip-10 expression by IFN-gamma and IL-1beta occurs even after the activation of classical NF-kappaB has returned to basal levels. Therefore, IKK-beta has a novel role in IFN-gamma-dependent activation of chemokine gene expression through its activation of IRF1 and p65.

39: The Journal of infectious diseases, 2009 Nov 23, 57(22)
Early Interferon-gamma Response against Plasmodium falciparum Correlates with Interethnic Differences in Susceptibility to Parasitemia between Sympatric Fulani and Dogon in Mali.

[Abstract]Introduction. Interethnic differences in susceptibility to malaria provide a unique opportunity to explore immunological correlates of protection. The Fulani of Sahelian Africa are known for their reduced susceptibility to Plasmodium falciparum, compared with surrounding tribes, yet the immunology underlying this is still poorly understood. Methods and Results. Here, we show that mononuclear cells from Fulani elicit >10-fold stronger interferon (IFN)-gamma production following a 24-h in vitro coincubation with asexual parasites than cells from sympatric Dogon. This response appears to be specific for P. falciparum among a panel of other human pathogens and is independent of the lower number of regulatory T cell counts present in Fulani. IFN-gamma responses in both tribes were inversely correlated with peripheral parasite density as quantified by nucleic acid sequenced-based amplification, but responses of Fulani remained significantly stronger than those of Dogon after adjustment for concurrent parasitemia, suggesting that hard-wired immunological differences underlie the observed protection. Conclusions. These results underscore the value of early IFN-gamma responses to P. falciparum as a correlate of anti-parasite immunity, not only in this setting but also in the wider context of malaria, and support the development of malaria vaccines aimed at inducing such responses.

40: The European respiratory journal : official journal of the European Society for Clinical Respiratory Physiology, 2009 Nov 19, 57(22)
Effect of TB treatment on T-cell interferon-{gamma} responses to M. tb-specific antigens.

[Abstract]The hypothesis that T-cell interferon-gamma responses to Mycobacterium tuberculosis-specific antigens decline as disease activity diminishes with TB treatment has generated interest in the interferon-gamma release assays (IGRAs) as treatment monitoring tools. We studied the effect of TB treatment on these responses as measured by the QuantiFERON-TB Gold In-tube(R) (QFT-IT) and T-SPOT.TB(R).275 sputum culture-positive, HIV-uninfected pulmonary TB patients were tested with QFT-IT and T-SPOT.TB at baseline, treatment completion and six months thereafter. The QFT-IT was also performed at the end of the intensive phase. The time-treatment effect on the qualitative and quantitative IGRA results was determined.There were significant declines in the positivity rates and quantitative results of both IGRAs with treatment (p<0.0001). The QFT-IT positivity rate was significantly lower than the T-SPOT.TB (p<0.0001). The test reversion rate was significantly different for the two assays (13.9% for T-SPOT vs 39.2% for QFT-IT, p<0.0001). 79% and 46% tested positive with T-SPOT.TB and QFT-IT respectively at six months post-treatment completion. The kinetics of the quantitative responses was not significantly different between subjects with and without risk factors for disease relapse.That a substantial proportion of patients remained test-positive after TB treatment would suggest a limited role of IGRAs as treatment monitoring tools.

41: Journal of clinical microbiology, 2009 Nov 18, 57(22)
Diagnosis of congenital toxoplasmosis using whole-blood interferon-{gamma} release assay.

[Abstract]Congenital toxoplasmosis at birth is generally subclinical but infected infants are at risk of developing ocular lesions. Diagnosis at birth relies mainly on serological tests. Cell- mediated immunity plays the major role in resistance to infection but is not routinely investigated for diagnostic purposes. Here we describe a simple test based on the response to interferon-gamma (IFN-gamma) after stimulation of whole blood by crude parasitic antigens. One millilitre of heparinized blood was centrifuged; plasma was kept for routine serological tests and pellets were resuspended in culture medium. After 24 h of culture in the presence of crude Toxoplasma gondii antigen, the cells were centrifuged and IFN-gamma was assayed in the supernatant. In 62 infants under 1 year of age born to mothers who were infected during pregnancy, the sensitivity and specificity were 94% (16/17) and 98% (44/45), respectively. The false negative was in a treated baby who turned positive after withdrawal of treatment. The false positive was observed in 3-month-old baby. In a cohort of 124 congenitally infected patients aged between 1 and 30 years, the sensitivity of the test was 100%. We present a simple test based on IFN-gamma secretion to assess cell-mediated immunity in toxoplasmosis. As only 1 mL of blood is required to investigate humoral and cellular immunity, our assay is well adapted for the study of congenital toxoplasmosis in infants. Using purified antigens or recombinant peptides might improve the test performances.

42: British journal of pharmacology, 2009 Nov 17, 57(22)
The benzoxathiolone LYR-71 down-regulates interferon-gamma-inducible pro-inflammatory genes by uncoupling tyrosine phosphorylation of STAT-1 in macrophages.

[Abstract]Background and purpose: Benzoxathiolone derivatives have shown anti-inflammatory and immunomodulatory potential in acne and psoriatic disorders. However, little is known about the molecular basis for these pharmacological effects. In this study, we decided to investigate the anti-inflammatory actions of a benzoxathiolone derivative LYR-71, 6-methyl-2-propylimino-6,7-dihydro-5H-benzo[1,3]oxathiol-4-one, in interferon (IFN)-gamma-activated macrophages. Experimental approach: RAW 264.7 macrophages or primary macrophages, derived from bone marrow of C3H/HeJ mice, were stimulated with IFN-gamma in the presence of LYR-71. Nitric oxide (NO) or chemokine production was measured by Griess reaction or enzyme-linked immunosorbent assay. RAW 264.7 cells were used to examine the molecular mechanisms of LYR-71 in modulating IFN-gamma-induced inflammatory responses. Key results: LYR-71 down-regulated IFN-gamma-induced transcription of inducible NO synthase, IFN-gamma-inducible protein-10 and the monokine induced by IFN-gamma genes in macrophages. This effect was mediated by uncoupling tyrosine phosphorylation of the signal transducer and activator of transcription (STAT)-1 in response to IFN-gamma. LYR-71 directly inhibited the in vitro catalytic activity of Janus kinase (JAK)-2. Further, the inhibitory actions of LYR-71 on IFN-gamma-induced STAT-1 phosphorylation and NO production were consistently abolished in the presence of peroxyvanadate, implying another target dependent on protein tyrosine phosphatase. Conclusions and implications: Taken together, LYR-71 could restrain IFN-gamma-induced inflammatory responses through uncoupling the tyrosine phosphorylation of STAT-1, an activation index of JAK-STAT-1 signalling, in macrophages. These results may provide a molecular mechanism underlying anti-inflammatory actions shown by benzoxathiolone derivatives.

43: Annals of the rheumatic diseases, 2009 Nov 16, 57(22)
Interferon-gamma gene polymorphisms associated with susceptibility to systemic lupus erythematosus.

[Abstract]OBJECTIVE: Interferon-gamma (IFNG) is a type II interferon playing diverse roles in innate and adaptive immune systems. Elevated expression of IFNG has been associated with systemic lupus erythematosus (SLE). This study aimed to examine association of IFNG polymorphisms with SLE susceptibility. METHODS: Five tag single-nucleotide polymorphisms (SNPs) and eight variations in all known regulatory sequences affecting IFNG expression within and around IFNG were genotyped in 1759 unrelated Korean subjects. SLE susceptibility association was assessed by comparing 742 SLE patients and 1017 unaffected controls using multivariate logistic regression analysis with adjustment for age and gender. RESULTS: SLE susceptibility association was significant with rs2069705 in the promoter (adjusted OR 2.27, p = 0.0024) and marginal with rs3181032 in the promoter (p = 0.037), rs2430561 in intron 1 (p = 0.022) and rs2069718 in intron 3 (p = 0.026) in a recessive genetic model. Five other SNPs showed no association and four other variations were not polymorphic. CONCLUSION: Several SNPs in IFNG are associated with SLE susceptibility, and the risk allele of an associated SNP (rs2430561) located in an NF-kappaB binding site has elevated IFNG expression versus the non-risk allele, supporting that elevated IFNG expression is associated with increased SLE susceptibility.

44: Journal of agricultural and food chemistry, 2009 Nov 25, 57(22)
FIP-fve Stimulates Interferon-Gamma Production via Modulation of Calcium Release and PKC-alpha Activation.

[Abstract]Fungal immunomodulatory protein, FIP-fve, has been isolated from Flammulina velutipes, and its immunomodulatory effects are believed to be associated with the enhanced activation of IFN-gamma-releasing Th1 cells. However, the mechanisms of FIP-fve-mediated signal transduction in the regulation of interferon-gamma (IFN-gamma) gene expression in human peripheral blood mononuclear cells (PBMCs) are still poorly understood. Using fluo-3 AM, we found that FIP-fve induces a rapid elevation in calcium concentration. ELISA, RT-PCR and Western blot assays demonstrated significant increases in the production and mRNA expression of IFN-gamma and protein kinase C-alpha (PKC-alpha) activation in activated PBMCs, which were abolished by EGTA, nifedipine and GO6976. In conclusion, Ca(2+) release and PKC-alpha activation are required for IFN-gamma production induced by FIP-fve in PBMCs.

45: Biochemical and biophysical research communications, 2009 Nov 11, 284(46)
IL-11 Expression in Retinal and Corneal Cells is Regulated by Interferon-gamma.

[Abstract]Interleukin-11 (IL-11) is an anti-apoptotic, anti-inflammatory cytokine with hematopoietic potential. The expression and protective actions of IL-11 have not been explored in the eye. The expression of IL-11 in primary cultures of human retinal pigment epithelial (HRPE) and human corneal fibroblast (HCRF) cells were evaluated in these studies. Constitutive secretion of IL-11 was not observed in either HRPE or HCRF. TNF-alpha +IL-1 induced IL-11 secretion and this production was inhibited by NFkappaB pathway inhibitors. IFN-gamma significantly inhibited TNF-alpha and IL-1 induced IL-11 secretion and inhibitors of JAK-STAT pathway reversed this inhibition. TGF-beta induced IL-11 secretion that was blocked by TGF-beta receptor 1 inhibitor but not by IFN-gamma. RT-PCR analysis confirmed the effects of IL-1, TNF-alpha, IFN-gamma and TGF-beta on IL-11 secretion at mRNA levels. Our results demonstrate that IL-11 is dramatically up regulated in retina and cornea cells and that IFN-gamma is a physiological inhibitor of IL-11 expression.

46: The Journal of infectious diseases, 2009 Nov 12, 284(46)
Age-Related Increase in the Frequency of CD4(+) T Cells That Produce Interferon-gamma in Response to Staphylococcal Enterotoxin B during Childhood.

[Abstract]Background. The susceptibility of infants to infections is well defined clinically, and immunologic abnormalities have been described. Immune maturation is complex, however, and the interval during which changes occur during childhood has not been identified. Methods. To assess age-related differences in the CD4(+) T cell responses, we evaluated the frequency of CD4(+) T cells that produced interferon (IFN) gamma in response to staphylococcal enterotoxin B (SEB) stimulation in 382 healthy infants and children (2 months to 11 years of age) and 66 adults. Flow cytometry was used to assess SEB-induced CD69 and CD40 ligand (CD40-L) expression and IFN-gamma production by CD4(+) and CD45RO(+)CD4(+) T cells. Results. CD69 and CD40-L expression by CD4(+) and CD45RO(+)CD4(+) T cells were similar to adult levels from infancy, but the frequency of activated T cells that produced IFN-gamma remained lower than adult responses until children were 10 years of age. Conclusions. These observations indicate that the IFN-gamma response of CD4(+) T cells to SEB remains limited for a much longer interval than was reported elsewhere, extending to the second decade of life. Observed differences in CD45RO(+)CD4(+) T cell function indicate that CD4(+) T cells with the same phenotypes do not possess equivalent functional capabilities.

47: Infection, 2009 Nov 10, 284(46)
Interferon-Gamma Release Assay in the Ascites: Early Hint for Diagnosis of Abdominal Tuberculosis.

[Abstract]We report on a 20-year-old woman with abdominal tuberculosis. Standard microbiological examination of ascites showed no acid-fast bacilli (AFB), and analysis for the Mycobacterium tuberculosis (MTB)-complex DNA by PCR was negative. However, the interferon-gamma release assay (IGRA) of the ascites was positive after specific stimulation with mycobacterial antigens (ESAT-6/CFP-10/TB7.7[p4]), indicating an infection with MTB. The diagnosis of tuberculosis was later confirmed by histology, MTB culture, and PCR analysis of MTB-complex DNA in tissue samples taken during laparoscopy. Thus, the IGRA of ascites may guide the decision to start active treatment for tuberculosis.

48: Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology, 2009 Nov 6, 105(10)
Interferon-gamma and pulmonary macrophages contribute to the mechanisms underlying prolonged airway hyperresponsiveness.

[Abstract]Background Airway hyperresponsiveness (AHR) in asthmatics includes a variable component that persists following an allergen challenge. This may be dissociated from inflammatory cell recruitment, implying a role for resident pulmonary cells in regulating the response. Objective Using improved methods of assessing AHR in a mouse model of allergic airway disease, to investigate the basis of the development of prolonged AHR. Method BALB/c mice were systemically sensitized and then challenged with aerosolized ovalbumin (OVA). Airway and tissue responsiveness were measured at baseline and at 1 day, and 1, 2 and 3 weeks after the last OVA challenge. Inflammatory cell numbers in BALF and levels of mRNA for eotaxin-1 and -2, IFN-gamma, IL-5 and -13 in the lung were measured at each time-point. In further experiments, the roles of IFN-gamma and of CCR3(+) and CD4(+) cells in the development of prolonged AHR were assessed by blockade or depletion with monoclonal antibodies. The role of pulmonary macrophages was assessed by selective chemical depletion of these cells. Results Airway responsiveness was increased above baseline at 1 day after the last OVA challenge, and this was sustained for 1 week. In contrast, tissue-specific responsiveness was only significantly increased above baseline at 1 day. Development of prolonged AHR was inhibited by neutralization of IFN-gamma or by depletion of pulmonary macrophages, but not by depletion of either CD4(+) T cells or CCR3(+) eosinophils. Conclusion An interaction between IFN-gamma and pulmonary macrophages contributed to the prolongation of airway hyperresponsiveness. In contrast, T cells and eosinophils did not contribute to prolongation of AHR. These findings emphasize the importance of the innate host response in the development of manifestations of asthma, as well as its potential relevance as a target for therapeutic intervention.

49: Clinical cancer research : an official journal of the American Association for Cancer Research, 2009 Nov 3, 27(47)
Production of Interferon-{gamma}-Inducible Protein-10 and Its Role as an Autocrine Invasion Factor in Nasal Natural Killer/T-Cell Lymphoma Cells.

[Abstract]PURPOSE: Nasal natural killer (NK)/T-cell lymphoma is associated with Epstein-Barr virus and has poor prognosis because of local invasion and/or multiple dissemination. Recently, the role of chemokines/chemokine receptors in tumor proliferation and invasion has been shown. In this study, we examined whether the specific chemokines were related to the tumor behaviors in nasal NK/T-cell lymphoma. EXPERIMENTAL DESIGN: A chemokine protein array was used to examine specific chemokines produced by SNK-6 and SNT-8 (Epstein-Barr virus-positive nasal NK/T-cell lymphoma lines). The expression of interferon gamma-inducible protein 10 (IP-10) and the IP-10 receptor CXCR3 was investigated by ELISA and flow cytometry. Cell growth and invasion were assessed by the MTT and Matrigel invasion assays, respectively. Immunohistologic staining and ELISA were used to examine IP-10 expression in biopsies and sera from patients, respectively. RESULTS: IP-10 was specifically produced by SNK-6 and SNT-8. Moreover, CXCR3 was expressed on the NK cell lines. Functionally, IP-10 did not affect cell proliferation but enhanced cell invasion. In biopsy samples, IP-10 and CXCR3 expressions were detected in the lymphoma cells. Serum IP-10 levels in the patients were much higher than those of healthy controls and the levels were decreased during the complete remission phase after treatments. CONCLUSIONS: These results suggest that IP-10 may play an important role in cell invasion in nasal NK/T-cell lymphoma through an autocrine mechanism. (Clin Cancer Res 2009;15(22):6771-9).

50: Medical mycology : official publication of the International Society for Human and Animal Mycology, 2009 Nov 3, 27(47)
Vertebral osteomyelitis due to Aspergillus fumigatus in a patient with chronic granulomatous disease successfully treated with antifungal agents and interferon-gamma.

[Abstract]We report a case of invasive aspergillosis due to Aspergillus fumigatus involving the cervical and thoracic vertebrae and upper mediastinum of a 17 year-old Saudi male with chronic granulomatous disease (CGD). The patient did not respond to a long course of liposomal amphotericin B but did to surgical drainage and a combination of caspofungin and itraconazole with subsequent suppression with oral voriconazole. Fourteen months after the start of therapy, the patient had anterior dislocation of T2 thoracic vertebra with cord transection and quadriplegia. He was then treated intravenously with liquid itraconazole and interferon-gamma. The patient made a remarkable recovery over a 2-year period and was eventually able to walk independently. Thus, a combination of antifungals and interferon-gamma may have resulted in the positive outcome in this case.

51: Archives of disease in childhood, 2009 Nov 20, 57(22)
Comparison of Interferon-gamma release assays and Tuberculin Skin Test in predicting active tuberculosis (TB) in children in the UK- a Paediatric TB Network Study.

[Abstract]BACKGROUND: The value of interferon-gamma release assays (IGRA) to diagnose active tuberculosis (TB) in children is not established, but these assays are being widely used for this purpose. We examined the sensitivity of commercially available IGRA to diagnose active TB in children in the UK compared with the tuberculin skin test (TST). METHODS: We established a paediatric tuberculosis network (PTBNET-UK) and conducted a retrospective analysis of data from children investigated for active TB at six large UK paediatric centres. All centres had used TST and at least one of the commercially available IGRA (T-Spot.TB or Quantiferon-Gold in Tube) in the diagnostic work up for active TB. Data were available from 333 children aged 2months to 16 years. We measured the sensitivity of TST and IGRA in definite (culture confirmed) and probable TB in children, agreement between TST and either IGRA and their combined sensitivity. RESULTS: Of 333 children, 49 fulfilled the criteria of definite TB and 146 had probable TB. Within the definite cohort, TST had a sensitivity of 82%, Quantiferon-Gold in tube (QFT-IT) had a sensitivity of 78% and T-Spot.TB of 66%. Neither IGRA performed significantly better than a TST with a cut-off of 15 mm. Combining results of TST and IGRA increased the sensitivity to 96% for TST plus T-Spot.TB and 91% for TST plus QFG-IT in the definite TB cohort. CONCLUSIONS: A negative IGRA does not exclude active TB disease, but a combination of TST and IGRA increases the sensitivity for identifying children with active TB.

52: The Pediatric infectious disease journal, 2009 Dec, 28(12)
GENE POLYMORPHISM OF IFNG +874 T/A AND TLR4 +896 A/G AND RECURRENT INFECTIONS AND WHEEZING IN TODDLERS WITH HISTORY OF BRONCHIOLITIS.

[Abstract]Cytokine and TLR4 polymorphisms and their association with the infection history of 129 children hospitalized for bronchiolitis during the first 6 months of life were analyzed. The carriers of IFNG +874 T/A allele A had fewer infections and use of inhaled corticosteroids and the carriers of TLR4+896 A/G allele G were more likely to need tympanostomy than noncarriers.

53: Human gene therapy, 2009 Nov 17, 57(22)
Therapeutic Vaccination with an Interleukin-2-Interferon-gamma-Secreting Allogeneic Tumor Vaccine in Patients with Progressive Castration-Resistant Prostate Cancer: A Phase I/II Trial.

[Abstract]Abstract Immunotherapy with whole cell cancer vaccines has been tested in various tumor types. This study investigated the safety profile and antitumor activity of an allogeneic prostate carcinoma cell line, LNCaP, expressing recombinant human interleukin-2 and human interferon-gamma. Thirty HLA-A*0201-matched patients with progressive, castration-resistant prostate cancer received four intradermal injections on days 1, 15, 29, and 92, and then every 90 days, as long as no tumor progression occurred. Three patients received a dose level of 7.5 million cells, and 27 patients received 15 million cells per injection. The primary study criteria were safety and the difference in prostate-specific antigen doubling time (PSA-DT), determined in the pretreatment phase (before the start of vaccination) and in the trial treatment phase (during vaccination). No dose-limiting or autoimmune toxicity was seen. During vaccination there was a significant prolongation of the PSA-DT compared with the prevaccination period (prolongation from 63 to 114 days; p < 0.01; intention to treat). In addition, results showed a period of PSA stabilization of at least 12 weeks, together with stable bone scans in 12 of 30 patients, and 3 patients sustained a >50% decrease in PSA versus baseline. The median overall survival time from first vaccination was 32 months (mean value, 34 months). Immune monitoring revealed T cell stimulation in the majority of patients. This vaccine strategy was found to be safe and well tolerated and was accompanied by prolongation of PSA-DT. The results of this trial warrant clinical development of this vaccine.

54: Glia, 2010 Jan 15, 58(2)
STAT1/IRF-1 signaling pathway mediates the injurious effect of interferon-gamma on oligodendrocyte progenitor cells.

[Abstract]Interferon-gamma (IFN-gamma) is a pleiotropic cytokine that is critically involved in the pathogenesis of inflammatory demyelinating diseases. There is strong evidence that IFN-gamma can function as a distinct and independent injurious factor to oligodendrocyte progenitor cells (OPCs). The intracellular signaling pathways leading to OPC death, however, remain poorly understood. In this study, we examined IFN-gamma signaling in OPCs in relation to cell death in vitro. Using expression knock-down and forced overexpression methods, we directly demonstrated the role of signal transducer and transcription activator 1 (STAT1) and interferon-regulated factor 1 (IRF-1) in IFN-gamma- induced OPC death. In addition, our study identified two proapoptotic genes, caspase 1 and double-stranded RNA-dependent protein kinase (PKR), whose expression was upregulated by IFN-gamma and transcriptionally controlled by IRF-1. The conclusion of this study is that STAT1 and IRF-1 function as components of the signaling pathway that mediates IFN-gamma-induced OPC death. (c) 2009 Wiley-Liss, Inc.

55: Veterinary immunology and immunopathology, 2009 Dec 15, 132(2-4)
Secretory expression of porcine interferon-gamma in baculovirus using HBM signal peptide and its inhibition activity on the replication of porcine reproductive and respiratory syndrome virus.

[Abstract]The gene sequence encoding mature porcine interferon-gamma (PoIFN-gamma) fused with a C-terminal 6x histidine tag was cloned into the baculovirus pFastBac Dual vector of the Bac-to-Bac Baculovirus expression system under the control of PH promoter. The authentic signal sequence of porcine interferon-gamma was substituted with the honeybee melittin (HBM) signal sequence, and expressed in insect cells. The recombinant proteins were detected by SDS-PAGE and immunofluorescence assay. The nickel affinity column purified recombinant porcine interferon-gamma with HBM signal peptide (rPoIFN-gammaH) was shown to be a 19kDa protein as confirmed by Western blot analysis. The recombinant PoIFN-gammaH was shown to have cytokine activity, inhibiting the cytopathic effect of vesicular stomatitis virus (VSV) in PK-15 cells at about 1.07x10(6)U/mL. The 2(-7) dilution of the rPoIFN-gammaH in culture supernatant protected the MARC-145 cells from the cytopathic effect caused by 100TCID(50) of porcine reproductive and respiratory syndrome virus.

56: Anatomical record (Hoboken, N.J. : 2007), 2009 Feb 26, 292(3)
Expression and Identification of Immunological Activities of the HIV-gp120N-Human Interferon Gamma Fusion Protein.

[Abstract]The human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein gp120 is a vaccine immunogen that has been studied extensively. To enhance the immune response of cells against HIV-1 gp120, we tested the coexpression of gp120N with interferon-gamma (IFN-gamma) as an immune adjuvant. Two recombinant prokaryotic plasmids were constructed: the pET44b-HIV-1-gp120N plasmid construct carried the HIV-1 gp120N gene (pET44-gp120N), whereas the pET44b-HIV-1-gp120N-IFN-gamma plasmid construct carried a fusion gp120N-IFN-gamma gene (pET44b-gp120N-IFN-gamma). Target protein expression was achieved in E. coli BL21 (DE3) cells by chemical induction. To test the immunological activity of the proteins, mice were injected with a control, gp120N, or the fusion gp120N-IFN-gamma protein. The serum and spleen cells of the mice were collected for immunological detection. Results showed that specific T lymphocyte proliferation and the expression of the Th1-type cytokines (IL-2 and IFN-gamma) were higher in the gp120N-IFN-gamma group than the other two groups (P < 0.05). No difference was observed in the expression levels of the Th2-type cytokines (IL-4 and IL-10; P > 0.05). These results suggest that IFN-gamma plays a prominent role as an immune adjuvant when coexpressed with HIV-1 gp120N. IFN-gamma enhances the specific cell immune response of mice against HIV-1 gp120. Anat Rec, 292:381-386, 2009. (c) 2009 Wiley-Liss, Inc.

57: Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 2009 Feb 26, 292(3)
Detection and Prediction of Active Tuberculosis Disease by a Whole-Blood Interferon-gamma Release Assay in HIV-1-Infected Individuals.

[Abstract]Background. The sensitivity of whole-blood interferon-gamma release assays to detect or predict active tuberculosis in individuals infected with human immunodeficiency virus type 1 (HIV-1) has as yet not been determined. Methods. In this prospective, longitudinal, single-center study, 830 HIV-1-infected patients underwent testing with the QuantiFERON-TB Gold In-Tube (QFT-GIT) assay. Clinical screening for active tuberculosis was performed at least every 3 months for a median follow-up time of 19 months. Results. At baseline, the QFT-GIT assay yielded positive or indeterminate results in 44 (5.3%) and 47 (5.7%) of the 830 patients, respectively. A positive QFT-GIT assay result occurred at significantly higher frequencies among black individuals than among white individuals (odds ratio, 4.84; 95% confidence interval, 2.25-9.97; [Formula: see text]), among patients from Africa than among patients from Austria (odds ratio, 6.57; 95% confidence interval, 2.99-14.25; [Formula: see text]), and among patients from high-prevalence countries than among patients from low-prevalence countries (odds ratio, 5.86; 95% confidence interval, 2.41-13.44; [Formula: see text]). In patients with indeterminate QFT-GIT assay results, both median actual and nadir CD4(+) T cell counts were significantly lower than in patients with interpretable QFT-GIT assay results ([Formula: see text]). At the time of baseline QFT-GIT screening, active tuberculosis was found in 7 (15.9%) of 44 individuals with a positive result and in 1 (0.1%) of 739 patients with a negative result. During the follow-up period, however, progression to active tuberculosis occurred exclusively in patients with a positive QFT-GIT assay result, at a rate of 8.1% (3 of 37 patients; [Formula: see text]). Collectively, the sensitivity of the QFT-GIT assay for active tuberculosis was 90.9% (95% confidence interval, 62.3%-98.4%). Conclusions. Our results suggest that the QFT-GIT assay may be a sensitive tool for the detection and prediction of active tuberculosis in HIV-1-infected individuals.

58: Epidemiology and infection, 2009 Feb 26, 59 Suppl 6(3)
Prospective evaluation of latent tuberculosis with interferon-gamma release assays in drug and alcohol abusers.

[Abstract]SUMMARYIn vitro tests have been developed for the diagnosis of tuberculosis (TB) infection. The objective was to analyse latent TB infection in drug and alcohol abusers through two interferon-gamma techniques. One hundred and thirty-nine patients were admitted between February 2006 and May 2007. Mean age was 39.8 years [31% HIV positive]. The enzyme immunoassay (EIA) and enzyme-linked immunospot (ELISPOT) interferon-gamma assays were positive in 34% of patients with an agreement of 83% (kappa=0.63). Tuberculin skin test (TST) was positive in 29% of patients and the agreement of TST with EIA and ELISPOT interferon-gamma assays was 85% (kappa=0.62) and 83% (kappa=0.57), respectively. Almost 50% of patients with history of TB had a positive in vitro test. In conclusion, we observed a high prevalence of latent TB and good agreement between the new in vitro tests that otherwise may continue to be positive long after developing TB disease.

59: Journal of physiology and pharmacology : an official journal of the Polish Physiological Society, 2008 Dec, 59 Suppl 6(3)
Interferon gamma production in the course of Mycobacterium tuberculosis infection.

[Abstract]It is not clear why some individuals with unknown predisposition develop tuberculosis, while others remain healthy in spite of heavy exposure. Interferon gamma (IFNgamma) is considered to be the key cytokine responsible for resistance to M. tuberculosis infection, as confirmed by increased susceptibility to mycobacterial infections in rare inherited defects in IL-12-IFNgamma axis. The aim of this study was to assess the IFNgamma production by peripheral blood lymphocytes from immunocompetent tuberculosis (TB) patients. The study group included 51 TB patients. In all cases, TB was confirmed by culture. Twenty healthy TB contacts were considered as control group. Commercially available ELISA-based assays were used to measure IFNgamma in the supernatant of whole blood cell cultures after stimulation with PWM (Phytolacca Americana), PHA (phytohemagglutynin), and PPD (purified protein derivative). No difference in IFNgamma secretion between the patients and control group was found when blood cells were stimulated by PWM or PHA. PPD-induced IFNgamma formation was higher in TB patients than in controls. The secretion of IFNgamma after non-specific stimulation varied in different clinical and radiological presentation of tuberculosis and it was lower in most advanced and extensive forms of the disease. It is unclear whether the difference in formation and release of IFNgamma is a primary or secondary phenomenon in the course of the disease.

60: Journal of physiology and pharmacology : an official journal of the Polish Physiological Society, 2008 Dec, 59 Suppl 6(3)
Pleural fluid adenosine deaminase and interferon gamma as diagnostic tools in tuberculosis pleurisy.

[Abstract]Several biological markers have been proposed to improve the efficacy of diagnosing tuberculous pleurisy. The study was undertaken to evaluate the accuracy of pleural fluid adenosine deaminase (ADA) activity and interferon-gamma (IFN-gamma) concentration in differentiating tuberculous pleural effusion (TPE) and nontuberculous pleural effusion (non-TPE). Ninety four patients (50 M and 44 F, mean age 60+/-18, range 18-95 years) with pleural effusion (PE) were studied. TPE was diagnosed in patients with: (i) positive pleural fluid or pleural biopsy culture or (ii) granulomas in the pleural biopsy specimen, after exclusion of other granulomatous diseases. Pleural fluid ADA activity was measured with the colorimetric method of Giusti, while IFN-gamma level was measured with ELISA. TPE was diagnosed in 28 patients. The non-TPE group consisted of 35 patients with malignant PE, 20 patients with parapneumonic effusion/pleural empyema, 5 with pleural transudate, and 6 with miscellaneous PE. The ADA activity and IFN-gamma concentration were significantly higher in TPE than in non-TPE (614.1+/-324.5 vs. 15.1+/-36.0 pg/ml, P<0.0001 and 75.1+/-39.1 vs. 11.0+/-16.6 U/l respectively, P<0.0001). The diagnostic sensitivity and specificity of IFN-gamma measurement (cut-off value of 75.0 pg/ml) were 100% and 98.5% respectively and were similar to those of ADA (100% and 93.9% at the cut-off value of 40.3 U/L). We conclude that pleural fluid ADA activity and IFN-gamma concentration are highly sensitive and specific markers of tuberculous pleurisy.

61: Journal of physiology and pharmacology : an official journal of the Polish Physiological Society, 2008 Dec, 59 Suppl 6(3)
Prevalence of latent tuberculosis infection in health care workers in Poland assessed by interferon-gamma whole blood and tuberculin skin tests.

[Abstract]Health care workers (HCWs) are at risk for developing active tuberculosis (TB). The prevalence of latent tuberculosis infection (LTBI) in this group is unknown in Poland, due largely to the problems associated with the interpretation of the tuberculin skin test (TST) in BCG immunized population. The goal of the present study was to assess the prevalence of LTBI in both clinical and non-clinical 155 HCWs (120 females and 35 males) and to compare the groups at different levels of risk. All participants were interviewed using a questionnaire and underwent interferon-gamma whole blood assay (Quantiferon-Tb-Gold) (QTF) and TST. The questionnaire provided information on possible risk factors for LTBI, including demographic and socioeconomic details, the presence of BCG scars, and the degree of occupational exposure. We found that the prevalence of LTBI among HCWs was, on average, 27.1%. A higher risk of acquiring LTBI was associated with certain work locations (TB lab workers--prevalence 50%, TB ward clinicians--34%, nurses--30%). The prevalence in analytical lab technicians was 20%, in administration staff was 15%. The HCWs with positive QTF test results were older and worked longer than those who had negative results. There was a significant correlation between the level of IFN-gamma and both age (P<0.001) and length of employment (P<0.01). The correlation between the diameter of skin test induration and the magnitude of the INF-gamma response also was significant (P<0.001). We conclude that HCWs are at increased risk of infection, suggesting that appropriate preventive strategies should be undertaken. IFN-gamma test is a useful tool in detecting LTBI cases in a country where BCG vaccination is a national policy.


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