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1: BMC medical genetics, 2010 Jun 19, 11(1)
Association of IL1A and IL1B loci with primary open angle glaucoma.

[Abstract]ABSTRACT: BACKGROUND: Recent studies suggest that glaucoma is a neurodegenerative disease in which secondary degenerative losses occur after primary insult by raised Intraocular pressure (IOP) or by other associated factors. It has been reported that polymorphisms in the IL1A and IL1B genes are associated with Primary Open Angle Glaucoma (POAG). The purpose of our study was to investigate the role of these polymorphisms in eastern Indian POAG patients. METHODS: The study involved 315 unrelated POAG patients, consisting of 116 High Tension Glaucoma (HTG) patients with intra ocular pressure (IOP) >21mmHg and 199 non-HTG patients (presenting IOP<20mmHg), and 301 healthy controls from eastern India. Genotypes were determined by polymerase chain reaction and restriction digestion for three single nucleotide polymorphisms (SNPs): IL1A (-889C/T; rs1800587), IL1B (-511C/T; rs16944) and IL1B (3953C/T; rs1143634). Haplotype frequency was determined by Haploview 4.1 software. The association of individual SNPs and major haplotypes was evaluated using chi-square statistics. The p-value was corrected for multiple tests by Bonferroni method. RESULTS: No significant difference was observed in the allele and genotype frequencies for IL1A and IL1B SNPs between total pool of POAG patients and controls. However, on segregating the patient pool to HTG and non-HTG groups, weak association was observed for IL1A polymorphism (-889C/T) where -889C allele was found to portray risk (OR= 1.380; 95% CI =1.041-1.830; p= 0.025) for non-HTG patients. Similarly, 3953T allele of IL1B polymorphism (+3953C/T) was observed to confer risk to HTG group (OR= 1.561; 95% CI =1.022-2.385; p= 0.039). On haplotype analysis it was observed that TTC was significantly underrepresented in non-HTG patients (OR= 0.538; 95% CI =0.356- 0.815; p= 0.003) while TCT haplotype was overrepresented in HTG patients (OR= 1.784; 95% CI =1.084- 2.937; p= 0.022) compared to control pool. However, after correction for multiple tests by Bonferroni method, an association of only TTC haplotype with non-HTG cases sustained (pcorrected = 0.015) and expected to confer protection. CONCLUSION: The study suggests that the genomic region containing the IL1 gene cluster influences the POAG pathogenesis mostly in non-HTG patients in eastern India. A similar study in additional and larger cohorts of patients in other population groups is necessary to further substantiate the observation.

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2: AIDS (London, England), 2009 Jun 1, 23(9)
IL1A alleles associate with a virological response to antiretroviral therapy in HIV patients beginning therapy with advanced disease.

[Abstract]Among Australians treated for advanced HIV disease, a suboptimal virological response was associated with allele 2 at interleukin 1 alpha (IL1A)-889 and IL1A+4845. This is confirmed and investigated using patients from AIDS Clinical Trials Group study 384 (n=532). Among non-African-American patients with CD4 T-cell count of< or = 100 cells/microl at baseline, IL1A+4845(*)TT was associated with poor virological outcome (P=0.03). Differences were smaller with higher baseline CD4 T-cell counts. IL1A-889 and IL1A+4845 were not in linkage disequilibrium in African-American patients, and IL1A+4845(*)T did not affect outcome.

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3: HIV medicine, 2008 Oct, 9(8)
The relationship between ApoE, TNFA, IL1a, IL1b and IL12b genes and HIV-1-associated dementia.

[Abstract]OBJECTIVES: Host genetic factors implicated in AIDS dementia complex (ADC) were studied. METHODS: DNA from ADC patients (n=56), unselected HIV-seropositive patients (n=112, 171, 185 and 204) and HIV-seronegative controls (n=204, 60, 60, 96 and 624) were typed for polymorphic loci in genes encoding tumour necrosis factor (TNF)-alpha, interleukin (IL)-1alpha, IL-1beta, IL-12 and Apolipoprotein E (ApoE). Diagnosis of ADC was based on neurological symptoms, signs and neuroimaging findings with other causes of dementia excluded. Patients selected had ADC stage > or =1 and CD4 counts of <500 cells/microL. RESULTS: Allele 2 of TNFA-308 was more common in ADC patients compared to HIV-positive or HIV-negative controls (P=0.005, 0.024). No other differences between ADC patients and control groups were significant. Meta-analyses confirmed these results. CONCLUSIONS: This study suggests that TNFA-308 allele 2 or an allele in linkage disequilibrium with this locus influences ADC.

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4: Tissue antigens, 2008 Jul, 72(1)
Complex effects of IL1A polymorphism and calpain inhibitors on interleukin 1 alpha (IL-1 alpha) mRNA levels and secretion of IL-1 alpha protein.

[Abstract]Alleles of IL1A-889(C>T) and IL1A+4845(G>T) are in linkage disequilibrium. Interleukin 1alpha (IL-1alpha) is produced as a precursor protein and cleaved at positions 117-118 by calpain, generating a mature protein for export. IL1A+4845 affects amino acids expressed at position 114 and hence may modulate calpain-mediated cleavage. We sought evidence for this mechanism in intact cells. Blood leukocytes from heterozygous donors released more IL-1alpha protein than cells from IL1A(1,1) donors, while release from IL1A(2,2) cells was variable. Genotype did not affect levels of IL-1alpha mRNA, so differential cleavage of the precursor is a feasible mechanism. However, genotype also had no effect on inhibition of IL-1alpha release by pretreatment with calpain inhibitors, and calpain inhibitors reduced IL-1alpha and tumor necrosis factor alpha mRNA levels. Hence, calpain inhibitors probably affect inhibition of signal transduction pathway rather than cleavage of IL-1alpha protein. As ratios of mu-calpain/calpastatin were lowest in heterozygous donors, genetically determined IL-1alpha levels may modulate transcription of calpain and calpastatin. This could reduce the impact of IL1A genotype on IL-1alpha secretion and amplify individual variation in levels generated in culture.

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5: Biology of reproduction, 2008 Jun, 78(6)
Gamendazole, an orally active indazole carboxylic acid male contraceptive agent, targets HSP90AB1 (HSP90BETA) and EEF1A1 (eEF1A), and stimulates Il1a transcription in rat Sertoli cells.

[Abstract]Gamendazole was recently identified as an orally active antispermatogenic compound with antifertility effects. The cellular mechanism(s) through which these effects occur and the molecular target(s) of gamendazole action are currently unknown. Gamendazole was recently designed as a potent orally active antispermatogenic male contraceptive agent. Here, we report the identification of binding targets and propose a testable mechanism of action for this antispermatogenic agent. Both HSP90AB1 (previously known as HSP90beta [heat shock 90-kDa protein 1, beta]) and EEF1A1 (previously known as eEF1A [eukaryotic translation elongation factor 1 alpha 1]) were identified as binding targets by biotinylated gamendazole (BT-GMZ) affinity purification from testis, Sertoli cells, and ID8 ovarian cancer cells; identification was confirmed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and Western blot analysis. BT-GMZ bound to purified yeast HSP82 (homologue to mammalian HSP90AB1) and EEF1A1, but not to TEF3 or HBS1, and was competed by unlabeled gamendazole. However, gamendazole did not inhibit nucleotide binding by EEF1A1. Gamendazole binding to purified Saccharomyces cerevisiae HSP82 inhibited luciferase refolding and was not competed by the HSP90 drugs geldanamycin or novobiocin analogue, KU-1. Gamendazole elicited degradation of the HSP90-dependent client proteins AKT1 and ERBB2 and had an antiproliferative effect in MCF-7 cells without inducing HSP90. These data suggest that gamendazole may represent a new class of selective HSP90AB1 and EEF1A1 inhibitors. Testis gene microarray analysis from gamendazole-treated rats showed a marked, rapid increase in three interleukin 1 genes and Nfkbia (NF-kappaB inhibitor alpha) 4 h after oral administration. A spike in II1a transcription was confirmed by RT-PCR in primary Sertoli cells 60 min after exposure to 100 nM gamendazole, demonstrating that Sertoli cells are a target. AKT1, NFKB, and interleukin 1 are known regulators of the Sertoli cell-spermatid junctional complexes. A current model for gamendazole action posits that this pathway links interaction with HSP90AB1 and EEF1A1 to the loss of spermatids and resulting infertility.

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6: Clinical and investigative medicine. M¨¦decine clinique et experimentale, 2007, 30(2)
Lack of association polymorphisms of the IL1RN, IL1A, and IL1B genes with knee osteoarthritis in Turkish patients.

[Abstract]PURPOSE: To examine whether polymorphisms of the interleukin 1 receptor antagonist (IL1RN), interleukin 1 alpha (IL1A) and interleukin 1 beta (IL1B) genes are markers of genetic susceptibility to knee osteoarthritis in Turkish patients. METHODS: One hundred and seven patients with knee osteoarthritis and 67 controls were studied. Three polymorphisms of IL1A, IL1B, and IL1RN genes were typed from genomic DNA. Allelic frequencies were compared between patients and control subjects. RESULTS: No significant differences were observed in genotype and allele frequencies of the IL1RN VNTR, IL1A+4845, IL1B+3953 genes polymorphisms between patients and controls. Furthermore, we did not detect any association genotypes of the polymorphisms with the clinical, radiological, and laboratory profiles of patients. CONCLUSIONS: The present study suggest that the IL1RN VNTR, IL1A+4845, IL1B+3953 genes polymorphisms are not genetic markers of susceptibility to knee osteoarthritis in Turkish patients, and are unrelated to the clinical, radiological, and laboratory characteristics of knee osteoarthritis.

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7: Immunogenetics, 2007 Jun, 59(6)
Contribution of single nucleotide polymorphisms of the IL1A gene to the cleavage of precursor IL-1alpha and its transcription activity.

[Abstract]We previously demonstrated the association of IL1A gene single nucleotide polymorphisms (SNPs) with susceptibility to systemic sclerosis (SSc) patients. In this study, we explored the effects of SNP on the transcriptional activity and processing of the precursor IL-1alpha (pre-IL-1alpha) in skin fibroblasts. Two kinds of promoter regions of the IL1A gene were prepared including C or T at -889, referred to C/IL1A and T/IL1A, and inserted into a luciferase reporter vector (pGL3). Skin fibroblasts were explanted from two SSc patients whose genomic DNA contained GG and TT genotypes at +4845 of the IL1A gene, respectively. Cell lysates were collected and reacted with various concentrations of calpain, and then the processing of pre-IL-1alpha was analyzed by Western blotting using monoclonal anti-IL-1alpha antibody. A SNP was determined by the allelic discrimination method using fluorescence-labeled Taq-Man probes. Significant differences in the luciferase activities were not detected between pGL3 (C/IL1A) and pGL3 (T/IL1A) in SSc fibroblasts. Calpain required a 100-fold higher concentration to process the pre-IL-1alpha containing Ala at the 114th amino acid than that to do containing Ser. The frequency of the GG genotype was significantly higher in SSc patients than that in healthy donors, whereas the frequency of TT genotype was significantly higher in RA patients than that in healthy donors. Our observation showed that the SNP at +4845 affected the enzymatic efficiency of the protease in cleaving pre-IL-1alpha. Pre-IL-1alpha with Ala, which was high in frequency in SSc patients, was more resistant to be cleaved by proteases in human sera than pro-IL-1alpha with Ser.

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8: Journal of periodontal research, 2007 Feb, 42(1)
The IL1A (-889) gene polymorphism is associated with chronic periodontal disease in a sample of Brazilian individuals.

[Abstract]BACKGROUND AND OBJECTIVE: It has been proposed that genotypes reflective of polymorphisms in cytokine genes can predispose individuals to disease by enhancing inflammatory processes. The C/T polymorphism at position -889 of the IL1A gene influences interleukin-1alpha expression, with the T allele inducing higher expression. The aim of this study was to evaluate the association of the IL1A (-889) gene polymorphism in Brazilian individuals with different clinical forms of periodontitis and severity of disease. MATERIAL AND METHODS: DNA was obtained from oral swabs of 163 Brazilian individuals and was amplified using the polymerase chain reaction (PCR). Products were submitted to digestion and were analyzed by electrophoresis to distinguish the C and T alleles. RESULTS: A significant difference in the genotype distribution was observed when comparing the chronic periodontitis group with the control group, evaluating only nonsmokers (chi-squared analysis = 9.91; p = 0.007), as well as when smokers were included (chi-squared analysis = 6.36; p = 0.04). Moreover, we observed a higher incidence of the T allele in the chronic periodontitis group (37.8%) when compared with the control group (18.4%) in nonsmokers (p = 0.006, odds ratio = 2.69, confidence interval = 1.27-5.68) and also when smokers were included (p = 0.03, odds ratio = 1.87, confidence interval = 0.98-3.56). No statistical difference was observed when the aggressive periodontitis group was compared with the control group. With regard to severity of disease, no statistical difference was observed. CONCLUSION: These data show an association of the IL1A (-889) polymorphism with chronic periodontitis in Brazilian individuals.

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9: Surgical neurology, 2006 Jan, 65(1)
Lack of association between the IL1A gene (-889) polymorphism and outcome after head injury.

[Abstract]BACKGROUND: Interleukin (IL) 1 is a proinflammatory cytokine that has been identified as an important mediator of neurodegeneration induced by ischemia or traumatic brain injury. Accumulating evidence to date has suggested that the major cytokine contributing to neurodegeneration after head injury is IL-1beta rather than IL-1alpha; however, there is no sufficient data regarding IL-1alpha in literature, and there may be an association between IL1A gene polymorphism and outcome after head injury. METHODS: We performed a prospective clinical study and included a recruited series of 71 patients who had head injury and were admitted to our neurosurgical unit. Severity of initial injury was assessed by the Glasgow Coma Scale. Outcome at 6 months after injury was assessed by means of the Glasgow Outcome Score. Interleukin 1alpha genotypes were determined from blood samples by standard methods. RESULTS: Of 40 patients with IL1A*2, 18 (45%) had an unfavorable outcome (dead, vegetative state, or severe disability) compared with 7 (22.5%) of 31 without IL1A*2 (P = .08). CONCLUSION: Our findings show that there is no genetic association between IL1A gene polymorphism and outcome after head injury. Further clinical studies should be designed to confirm and further evaluate these findings.

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10: The Journal of allergy and clinical immunology, 2005 Jul, 116(1)
Childhood cat exposure-related tolerance is associated with IL1A and IL10 polymorphisms.

[Abstract]BACKGROUND: Interleukin (IL) 1 is a proinflammatory cytokine that has been identified as an important mediator of neurodegeneration induced by ischemia or traumatic brain injury. Accumulating evidence to date has suggested that the major cytokine contributing to neurodegeneration after head injury is IL-1beta rather than IL-1alpha; however, there is no sufficient data regarding IL-1alpha in literature, and there may be an association between IL1A gene polymorphism and outcome after head injury. METHODS: We performed a prospective clinical study and included a recruited series of 71 patients who had head injury and were admitted to our neurosurgical unit. Severity of initial injury was assessed by the Glasgow Coma Scale. Outcome at 6 months after injury was assessed by means of the Glasgow Outcome Score. Interleukin 1alpha genotypes were determined from blood samples by standard methods. RESULTS: Of 40 patients with IL1A*2, 18 (45%) had an unfavorable outcome (dead, vegetative state, or severe disability) compared with 7 (22.5%) of 31 without IL1A*2 (P = .08). CONCLUSION: Our findings show that there is no genetic association between IL1A gene polymorphism and outcome after head injury. Further clinical studies should be designed to confirm and further evaluate these findings.

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11: Brain research bulletin, 2005 Jan 30, 64(6)
Stress-induced increases in hypothalamic IL-1: a systematic analysis of multiple stressor paradigms.

[Abstract]Exposure to stressors such as footshock, tailshock, and immobilization have been shown to induce hypothalamic IL-1 production, while other stressors such as restraint, maternal separation, social isolation, and predator exposure have no effect on hypothalamic IL-1 levels. This disparity of findings has led to considerable controversy regarding the ability of stressors to induce hypothalamic IL-1 expression. Thus, the goal of the following experiments was to examine hypothalamic IL-1 responses in adult male Sprague-Dawley rats following exposure to a diverse set of stressors. Our data indicate that exposure to 2h of restraint in a Plexiglas tube, glucoprivic challenge induced by administration of 2-deoxyglucose (2-DG), or insulin-induced hypoglycemia all fail to alter hypothalamic IL-1 levels despite robust activation of the pituitary-adrenal response. However, when restraint was administered on an orbital shaker or in combination with insulin-induced hypoglycemia, robust increases in hypothalamic IL-1 were observed. No effects of glucoprivic (2-DG) challenge were observed when combined with restraint, indicating some specificity in the hypothalamic IL-1 response to stress. We also provide a preliminary validation of the ELISA detection method for IL-1, showing that (a) Western blot analyses confirmed strong immunopositive banding at the apparent molecular weight of both mature IL-1beta and the IL-1beta prohormone, and (b) footshock led to a two-fold increase in mRNA for IL-1 in the hypothalamus as detected by RT-PCR. These data provide novel insight into the characteristics of a stressor that may be necessary for the observation of stress-induced increases in hypothalamic IL-1.

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12: Genes and immunity, 2004 Sep, 5(6)
Extended haplotypes and linkage disequilibrium in the IL1R1-IL1A-IL1B-IL1RN gene cluster: association with knee osteoarthritis.

[Abstract]The interleukin-1 gene cluster is a key regulator in a number of chronic disease processes. We explored the linkage between nine polymorphic loci in the IL1R1 promoter, eight in the IL1A-IL1B-IL1RN gene complex, and their association with osteoarthritis (OA), a common complex disease associated with low-level inflammation. Using 195 healthy controls, we identified eight novel polymorphisms in the IL1R1 exon 1A region. We found limited LD between IL1R1 and the IL1A-IL1B-IL1RN cluster, although LD within these two individual groups was high. To test association with knee OA, we genotyped 141 patients from Bristol (UK) at the 17 loci. IL1R1 promoter haplotypes showed no association with disease. However, within the IL1A-IL1B-IL1RN complex, we identified a common haplotype conferring a four-fold risk of OA (P=0.00043; Pc=0.0043) and one IL1B-IL1RN haplotype conferring a four-fold reduced risk (P=0.0036; Pc=0.029). To replicate these associations, we subsequently examined 163 knee OA patients from London. Here, the effects of the haplotypes were confirmed: the risk IL1A-IL1B-IL1RN haplotype conferred a two-fold risk of OA (P=0.02), and the protective IL1B-IL1RN haplotype conferred a five-fold reduced risk of OA (P=0.0000008). These results may help to explain the genome-wide scan linkage data and functional observations concerning association between IL-1 and OA.

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13: The Journal of allergy and clinical immunology, 2004 Mar, 113(3)
Epistatic effect of IL1A and IL4RA genes on the risk of atopy.

[Abstract]BACKGROUND: Several studies have demonstrated a linkage or association of the atopic phenotype with T-cell cytokine genes involved in the regulation of the TH1/TH2 balance (eg, IL4, IL13, and their common receptor, IL4RA). We have recently shown that polymorphism of the pro-inflammatory cytokine IL1A gene is strongly associated with atopy. OBJECTIVE: We now examined whether the polymorphisms of IL1A (G/T at +4845) and IL4RA (T/C at +22446) would show an epistatic effect on the risk of atopy. METHODS: Skin prick tests and gene polymorphism analyses were performed in a population-based sample of asthmatic and nonasthmatic subjects. RESULTS: Our results showed that in the nonasthmatic group the previously described elevated risk of atopy in noncarriers of allele T of IL1A (ie, having the genotype GG) was restricted to individuals who were also noncarriers of allele C of IL4RA (genotype TT). This finding applies to the general population of Finland, where 3.3% of adults are asthmatic. CONCLUSION: These data suggest that the IL1A and IL4RA genes show an epistatic effect on the risk of atopy.