Promoter Region Of IL-10 Gene Research Articles

Donor Genetic Predisposition to High Interleukin-10 Production Appears Protective against Acute Graft-Versus-Host Disease.

The persistence of graft-versus-host disease (GVHD) as the principal complication of allogeneic hematopoietic cell transplantation (HCT) demonstrates that HLA matching alone is insufficient to prevent alloreactivity. We performed molecular and functional characterization of 22 candidate cytokine genes for their potential to improve matching in 315 myeloablative, 10/10 HLA-matched donor−recipient pairs. Recipients of a graft carrying the -1082GG IL10 gene promoter region variant had a three-fold lower incidence of grade II−IV acute GVHD compared to IL10-1082AA graft recipients (SHR = 0.25, p = 0.005). This was most evident in matched unrelated donor (MUD) transplants, where the greatest alloreactivity is expected. IL10-1082GG transplants did not experience an increased incidence of relapse, and, consequently, overall survival was two-fold higher in IL10-1082GG MUD transplants (HR = 0.17, p = 0.023). Longitudinal post-transplant measurements demonstrated that -1082GG is a high-IL10-producing and -expressing genotype with attenuated CD8+ T-cell reconstitution. High post-transplant donor chimerism in T- and myeloid-cells (>95%) confirmed a predominant donor, rather than recipient, genotype effect on immune function and aGVHD. To date, this is the first study to report corroborating genome-to-cellular evidence for a non-HLA donor immunogenetic variant that appears to be protective against GVHD. The incorporation of IL10 variants in donor selection criteria and clinical-management decisions has the potential to improve patient outcomes.

Genetic polymorphisms of IL6 gene -174G>C and -597G>A are associated with the risk of COVID-19 severity.

Coronavirus disease-2019 (COVID-19) is pro-inflammatory disorder characterized by acute respiratory distress syndrome. Interleukin-6, a cytokine secreted by macrophages, which mediates an inflammatory response, is frequently increased and associated with the severity in COVID-19 patients. The differential expression of IL6 cytokine in COVID-19 patients may be associated with the presence of single nucleotide polymorphisms (SNPs) in regulatory region of cytokine genes. The aim of this study is to investigate the role of two promoter polymorphisms of the IL6 gene (-597G>A and -174G>C) with the severity of COVID-19. The study included 242 patients, out of which 97 patients with severe symptoms and 145 patients with mild symptoms of COVID-19. Genotyping of two selected SNPs, rs1800795 (-174G>C) and rs1800797 (-597G>A) of promoter region of IL6 gene, was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In our study, individuals with GC genotypes of IL6 (-174G>C) polymorphism showed significantly higher risk of severity [adjusted odds (OR) 3.86, p<.001] but we did not observe any association of COVID-19 severity with rs1800797 (-597G>A) polymorphism. The COVID-19 severity was significantly higher in individuals having 'C' allele of IL6 (-174G>C) polymorphism (p=.014). Linkage disequilibrium between rs1800795 (-174G>C) and rs1800797 (-597G>A) showed that individuals having AC* haplotype significantly association with COVID-19 severity (p=.034). Our results suggest that 'C' allele of rs1800795 (-174G>C) polymorphism of IL6 may be the risk allele for severity of COVID-19 in North Indian population.

Association between IL-10 rs1800872 Polymorphism in the Promoter Region of Human IL-10 Gene and Susceptibility to Rheumatoid Arthritis in the Patients in Southwest of Iran

Association between IL-10 rs1800872 Polymorphism in the Promoter Region of Human IL-10 Gene and Susceptibility to Rheumatoid Arthritis in the Patients in Southwest of Iran

Association between IL-18 and IL-6 gene polymorphisms and the risk of T1D in Egyptian children.

To test the involvement between IL-18 and IL-6 genetic polymorphisms and susceptibility to Type 1 diabetes (T1D). Single nucleotide polymorphisms (SNPs) at positions -607A/C and - 137G/C in IL-18 promoter region were examined by sequence specific primers-polymerase chain reaction (SSP-PCR) and position -174G/C in promoter region of IL-6 gene which analyzed by Mutagenically Separated PCR (MS-PCR) in 104 T1D participants and 114 controls. IL-18 -137GC and -137CC genotypes and -137C allele were significantly decreased in T1D subjects (P < 0.05), while -137GG genotype was insignificantly increased as compared to controls. A significant decrease was detected in haplotype -137C/-607C frequency in T1D participants compared with controls (OR = 0.04, P < 0.001). There was significant association between IL-18 -607 of (CC, AC and AA genotypes) in age at diagnosis, glycated hemoglobin (HbA1c) and higher body mass index (BMI) (P < 0.05). This study demonstrated that IL-18 gene promoter polymorphisms might be associated with susceptibility to T1D in Egyptian children. Individuals carrying CC genotype at position -137 of IL-18 promoter may be at a low risk of T1D progression. Additionally, the susceptible combination of IL-18 and IL-6 cytokine genes associated with T1D highlight their risk toward the disease. The online version contains supplementary material available at 10.1007/s40200-021-00763-w.

CRISPR/Cas9-Based Transcriptional Activation of Endogenous IL-10 Gene in the Donor Lung for Transplantation

Gene therapy of donor lungs has shown promise in lung transplantation (LTx). The discovery of the clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 system has advanced gene therapy by enabling precise and highly flexible modulation of endogenous genes. CRISPR-based gene modulation of a whole organ, which remains challenging, will allow us to optimize donor lungs by modulating any gene. To establish CRISPR-based epigenome editing for LTx, we started with activating the endogenous IL-10 gene, an anti-inflammatory gene, in rat lungs. We developed a specific vector targeting the IL-10 gene and tested in rat model. Adenoviral vectors expressing a Cas9 activator (Cas9-VPR) with or without a guide RNA (gRNA) designed to target the promoter region of the rat IL-10 gene were administered to Lewis rats via the airway at a dose of 5 × 108 PFU/rat. Rats were divided into three groups (n=6 each): 1) No gRNA - received adenovirus expressing Cas9-VPR alone; 2) gRNA - received adenovirus expressing Cas9-VPR and: 3) diluent - received buffer without any viruses. The lung was assessed for IL-10 gene expression and inflammation after 72 hours. The Cas9-VPR was detected in lung tissue of the gRNA group and the No gRNA group using immunohistochemistry (Fig A). The gRNA group showed increased IL-10 (5.85±2.05 vs 2.85±1.89 pg/mg protein, p=0.046) compared to the No gRNA group (Fig B) whereas TNFa and IFNg showed comparable level in both groups. Furthermore, the gRNA group showed improved oxygenation (587±41 vs 435±55 mmHg, p<0.0001, Fig C) and Wet/Dry ratio (5.04±0.14 vs 5.63±0.33, p=0.001, Fig D), demonstrating an anti-inflammatory effect of IL-10 gene activation on vector-induced inflammation in the donor lung. Using CRISPR/Cas9-based transcriptional modulation we have achieved targeted activation of the endogenous IL-10 gene in rat donor lungs. This proof of concept study opens the door to targeted gene modification of donor organs to improve outcomes after LTx.

Promoter polymorphisms in IL-6 gene influence pro-inflammatory cytokines for the risk of osteoarthritis

BackgroundInterleukin-6 (IL-6) gene regulates IL-6 levels, interplay of which has been found to influence pathophysiology of osteoarthritis (OA). Polymorphism within promoter region of IL-6 gene and its association with plasma levels of pro-inflammatory cytokines; IL-6, interleukin 1-beta (IL-1β) and tumor necrosis factor–alpha (TNF-α) remained to be investigated in Punjab region of India, where OA is highly prevalent. MethodsSix single nucleotide polymorphisms (SNPs) in the promoter region of IL-6 gene; rs1800795 (−174G/C), rs1800796 (−572G/C), rs1800797 (−597G/A), rs2069827 (−1363G/T), rs12700386 (−2954G/C) and rs10499563 (−6331G/T) were investigated by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 279 confirmed osteoarthritis patients and 287 controls. Plasma levels of pro-inflammatory cytokines; IL-6, IL-1β and TNF-α were measured by sandwich Enzyme Linked Immunosorbent Assay (ELISA). ResultsAllele frequency spectrum after adjusting the effect of systolic blood pressure (SBP), diastolic blood pressure (DBP), total cholesterol (TC), low density lipoprotein (LDL), triglycerides (TG) and body mass index (BMI) revealed that major allele G of rs1800795 and T of rs10499563 were significantly associated with increased risk of OA (P < 0.01) in all the three genetic models; co-dominant (OR 4.08 & 4.12, P < 0.001), recessive (OR 3.00 & 2.51, P < 0.001) and dominant (OR 2.56 & 3.09, P < 0.05). Major allele G of rs1800796 and rs1800797 was observed to enhance OA risk in recessive mode (OR 1.75, P < 0.001 & 1.62, P = 0.01 respectively). Disease risk analysis after adjusting the effect of confounders exposed a susceptibility haplotype GGGGCT, which increased the OA risk by 2.27 times (OR 2.27, 95%CI: 1.26–4.10, P = 0.009) and a protective haplotype CGAGGC which significantly reduced the OA risk (OR 0.47, 95%CI 0.27–0.92, P = 0.031). Both of these haplotypes manifested in the recessive mode of inheritance. Subjects who had one copy of the susceptibility haplotype had lower values of IL-6 (3.6 pg/ml) and IL-1β levels (3.2 pg/ml) than those who had 2 copies of it (4.4 pg/ml & 4.2 pg/ml respectively). IL-6 and IL-1β levels were observed to be negatively associated with protective haplotype CGAGGC (P < 0.05). Carriers of 1 copy of this haplotype showed decreased IL-1β levels than those who had none (1.00 pg/ml vs. 1.3 pg/ml respectively) which further decreased to 0.9 pg/ml in those subjects who carried two copies of protective haplotype. ConclusionThe present study discovered susceptibility (GGGGCT) and protective (CGAGGC) haplotypes within promoter region of IL-6 gene which influenced the plasma levels of IL-6 and IL-1β for the risk of osteoarthritis in the population of Punjab, India.

Association of interleukin-6 polymorphism (-634C/G) in the promoter region with diabetic nephropathy in type 2 diabetes mellitus

Background:Long term microvascular complications of DM includes: nephropathy, retinopathy, and neuropathy. Increased level of IL-6 and its polymorphism have been associated with diabetic nephropathy.The present study was designed to determine serum level of IL-6 and C/G polymorphism at − 634 position in the promoter region of IL-6 gene in patients with type 2 diabetes mellitus (T2DM) with and without nephropathy.&#x0D; Method: This comparative study was comprised of 164 subjects, divided into two groups of 82 subjects in each. The subjects who had diabetes for more than 5 years without nephropathy were labeled as group-I and the subjects who had diabetes for more than 5 years with nephropathy were labeled as group-II. IL-6 polymorphism was detected by RFLP and its level was determined by ELISA technique. Data was analyzed using SPSS 20.0.&#x0D; Results: In group-I, 14 (17.1%) patients had CC, 47 (57.3%) had CG and 21 (25.6%) had GG polymorphism. In group-II, 9 (11%) patients had CC, 69 (84.1%) had CG and 4 (4.9%) had GG polymorphism. Pearson’s correlation test determined significant association of IL-6 polymorphism (CC,CG,GG) in both the groups (p &lt;0.001). In patients of CC polymorphism, median levels of IL-6 was high in group-I (4.2) as compared to group-II (3.2),whereas in patients of CG polymorphism, median levels of IL-6 was high in group-II (3.8) as compared to group I (3.7) while in GG polymorphism, median levels of IL-6 was same in both the groups. On comparison of IL-6 polymorphism and level of IL-6 there was no statistically significant difference between the two groups (p1=0.304, p2=0.425 respectively).&#x0D; Conclusions: Statistically significant difference in the frequencies of allele (CC,CG,GG) between two groups is pointing towards the protective role of GG allele as it was predominant in T2DM patients without nephropathy.&#x0D; Bangladesh Journal of Medical Science Vol.18(4) 2019 p.741-747

CRISPR-Mediated IL-10 Gene Activation as a Novel Gene Therapeutic Strategy in Lung Transplantation

Purpose Lung transplantation (LTx) is life-saving treatment for end-stage respiratory failure. Our ultimate goal is to improve the outcome of LTx by upregulating the IL-10 gene, an anti-inflammatory and immune down-regulating cytokine, in the whole lung utilizing CRISPR-based technology. The CRISPR/Cas9 system, originally reported as a genome editing system, is being rapidly developed for various applications. CRISPR-mediated gene activation enables us to achieve specific and robust transcriptional activation of an endogenous gene by targeting the genomic DNA. The purpose of this study is to achieve CRISPR-mediated targeted IL-10 gene activation in vitro to generate a viral vector for future whole organ studies. Methods Seven guide RNAs (gRNA) were designed to target the promoter region of IL-10 gene of the rat genome (Figure A). First, each gRNA expressing plasmid was transfected together with a Cas9 activator (dSaCas9-VPR) expressing plasmid to a rat airway epithelial cell line (L2) and lung macrophage cell line (NR8383). Second, a single activation vector, which expressed both gRNA and Cas9 activator, was generated and transfected. The relative expression level of rat IL-10 was measured by qPCR. To determine the optimal viral vector for gene delivery to the lung, different adeno-associated (AAV) virus serotypes were tested in vitro using reporter genes. Results Both epithelial (Figure B) and macrophage cell lines demonstrated enhanced IL-10 gene expression when transfected with Cas9 activator and gRNA4 at 48h. Using a single vector, the gRNA expressing vector significantly increased IL-10 gene expression compared to mock (non-targeted) vectors in epithelial cells (Figure C). In viral vector testing, AAV1 and AAV6 showed effective transduction in macrophage cells. Conclusion Targeted IL-10 gene activation was achieved in both epithelial and macrophage cell lines. CRISPR-mediated IL-10 gene activation of the lung holds considerable promise as a potential gene therapy strategy.

A study of association between regulatory polymorphism in the IL-10 gene promoter region and acute viral hepatitis, and acute liver failure.

The level of inflammatory cytokine Interleukin (IL)-10 is increased in patients infected with hepatitis-related acute liver failure (ALF), and this was thought to be because of the regulatory polymorphism in the IL-10 gene promoter region. The present study was designed to analyze the possible association between IL-10 gene promoter polymorphism and acute viral hepatitis (AVH), and ALF. An attempt was made to quantify IL-10 levels at admission, during the hospital stay, and at the final outcome to study its relationship with liver injury among patients with AVH, ALF, and controls. The study included 40 patients each with ALF and AVH. IL-10 gene promoter polymorphism was detected by the PCR-RFLP method. Quantification of IL-10 was done using commercially available ELISA kits. The individuals with -592 AC, -819 TC, -1082 AA genotypes were found to have a significantly higher risk of ALF whereas those with -592 AA and - 819 CC polymorphism were found to be less susceptible. Individuals with - 819 CC were found to be more susceptible to AVH while those with -592 AA and -819 TT were less susceptible as compared to controls. Mean serum IL-10 at admission was significantly elevated in patients with ALF (38.4±11.3 pg/mL) as compared to patients with AVH (16.7±5.4pg/mL) and control population (8.3±3.6 pg/mL, p < 0.05). Regulatory polymorphism in the IL-10 gene promoter has a possible and significant association with severity and outcome in patients with AVH and ALF. Raised levels of IL-10 could be predictive of prognosis in patients with ALF.

Polymorphisms in the promoter region of IL10 gene are associated with virus etiology of infant bronchiolitis.

Bronchiolitis is the most common infection leading to hospitalization in infancy. Interleukin-10 (IL-10) is an anti-inflammatory cytokine, and in our previous study, IL10 gene rs1800896 (- 1082A/G) polymorphism was associated with viral etiology of infant bronchiolitis. The objective of this study was to evaluate the associations between IL10 single nucleotide polymorphisms (SNPs) at rs1800890 (- 3575A/T), rs1800871 (- 819C/T) or rs1800872 (- 592C/A) either alone or combined with the SNP at rs1800896 (- 1082G/A), and the etiology and severity of infant bronchiolitis. Data on four IL10 SNPs were available from 135 full-term infants, hospitalized for bronchiolitis at age less than 6months, and from 378 to 400 controls. Viral etiology was studied, and oxygen support, feeding support and the length of stay in hospital were recorded during bronchiolitis hospitalization. Infants with rhinovirus bronchiolitis had the IL10 rs1800890 variant AT or TT genotype less often (18.2%) than controls (63.3%, P = 0.03), and likewise, had the IL10 rs1800896 variant AG or GG genotype less often (27.3%) than controls (65.5%, P = 0.009). Twenty-eight infants with bronchiolitis had the variant-variant Grs1800896Trs1800890 haplotype, and none of them had rhinovirus infection. The IL10 rs1800871 or rs1800872 genotypes showed no associations with viruses. No association was found between any genotypes and bronchiolitis severity measures. IL10 rs1800890 and rs1800896 polymorphisms differed between infants with rhinovirus bronchiolitis and controls, but not between infants with respiratory syncytial virus bronchiolitis and controls.

Promoter haplotypes of interleukin-10 gene linked to cortex plasticity in subjects with risk of Alzheimer's disease.

The Alzheimer's disease (AD) aetiologic event is associated with brain inflammatory processes. In this study, we consider a haplotype of the IL-10 gene promoter region, − 1082A/− 819 T/− 592A (ATA haplotype), which is an additive and independent genetic risk factor for AD. Episodic memory change is the most striking cognitive alteration in AD. It remains unclear whether episodic memory networks can be affected by the ATA haplotype variant in amnestic mild cognitive impairment (aMCI), and if so, how this occurs. Thirty-nine aMCI patients and 30 healthy controls underwent resting-state functional magnetic resonance imaging. An imaging genetics approach was then utilized to investigate disease-related differences in episodic memory networks between the groups based on ATA haplotype-by-aMCI interactions. Gene-brain-behaviour relationships were then further examined. This study found that the ATA haplotype risk variant was associated with abnormal functional communications in the hippocampus-frontoparietal cortices, especially in the left hippocampal network. Moreover, these ATA haplotype carriers showed a distinct phase of hyperactivity in normal aging, with rapid declines of brain function in aMCI subjects when compared to non-ATA haplotype carriers. These findings added to the accumulating evidence that promoter haplotypes of IL-10 may be important modulators of the development of aMCI.

Genetic polymorphism of interleukin-10 (-A592C) among oral cancer with squamous cell carcinoma

ObjectiveInterleukin-10 (IL-10) is a pleiotropic cytokine with either immunosuppressive or immunostimulative activities. It has been reported that in cancer, the promoter region polymorphism of IL-10 (-A592C) alters both the expression and serum levels of this cytokine. In the present study, we have addressed the question as to whether the single nucleotide polymorphisms (SNPs) at positions −592 A/C in the IL-10 gene promoter, could predispose an individual to oral squamous cell carcinoma (OSCC). DesignWe analyzed the genotype of the IL-10 (-A592C) gene, in 250 histopathologically confirmed OSCC patients and similar number of healthy volunteers taken as controls, in an Indian population by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Allele and genotype frequencies were analyzed by the Student's t-test and the chi-squared test, and strength of associations by the odds ratio (OR) with 95% confidence intervals. ResultsThe genotype and allele distribution of IL-10 (-A592C) gene polymorphism was significantly different between OSCC cases and controls (genotype AA vs AC: OR 2.87; 95 % CI 1.50–5.48; p=0.0016 and AA vs CC: OR 4.08; 95 % CI 1.98–8.41; p=0.0002). The −592 C alleles were found to be significantly different among OSCC cases and controls (OR: 1.44, 95% CI: 1.12–1.85, p<0.0051). ConclusionsThe IL-10 gene promoter region (-592) A/C polymorphism is significantly associated with reduced risk of OSCC. The OSCC group had a significantly greater frequency of genotype AA as compared to control group.

Is the IL-6 -174G/C Gene Polymorphism Related to the Disease Severity Score in Turkish Children with Familial Mediterranean Fever?

Familial Mediterranean fever (FMF) is an autosomal recessively inherited disease characterized by recurrent self-limited attacks of fever accompanied by aseptic inflammation of serosal spaces, joints and skin, peritonitis, pleuritis, and arthritis. Clinical features differ according to genetics variants. The aim of this study was to identify relationship between IL-6 -174G/C gene polymorphisms and clinical features, disease severity score (DSS) and proteinuria in children diagnosed with FMF. In this study, 99 children who were followed-up in Gaziosmanpasa University Medical Faculty Department of Pediatrics and diagnosed with Familial Mediterranean fever according to Tel-Hashomer criteria were included. One hundred and fifty seven children who admitted to the hospital with any complain and found healthy included in control group. Genotyping was done for polymorphism in a promoter region of IL-6 gene (G/C at -174). The IL-6 -174G/C gene polymorphism and the clinical features of FMF, proteinuria, the DSS, and the healthy control group were investigated. Data for the clinical features were obtained retrospectively from the electronic records of patients. All of the genotyping of blood samples were done in Medical Genetic laboratory of Gaziosmanpasa University School of Medicine. The results revealed that the distribution of the genotypes and allele frequencies of the IL-6 -174G/C polymorphism were not significantly different between the FMF patients and the healthy controls. The IL-6 -174G/C polymorphisms did not affect proteinuria, the DSS, and the clinical features of FMF patients.

Polymorphism of IL-10 gene promoter region: association with T cell proliferative responses after acellular pertussis vaccination in adults.

Individual variation in immune responses is always encountered after vaccination. This phenomenon is also seen after acellular pertussis vaccination. The aim of this present study was to investigate whether single nucleotide polymorphisms (SNPs) in the IL-10 gene promoter region (rs1800890, rs1800896, rs1800871), IL-12B (rs2546890), IL-12RB1 (rs372889), IL-17A (rs2275913), and IL-23R (rs11209026) affect the immune responses after acellular pertussis vaccination. The T cell proliferative response was evaluated in 38 Finnish young adults who received a second booster dose of a vaccine combination of diphtheria, tetanus, and acellular pertussis, 10years after the previous booster. The response was evaluated with a proliferation assay in which vaccine antigens pertussis toxin (PT), filamentous hemagglutinin (FHA), and pertactin (PRN) were used for the stimulation, before and 1month after the second vaccination. Specific proliferation of peripheral blood mononuclear cells against pertussis antigens was affected by IL-10 SNP in the promoter region at position -1082 (A>G, rs1800896). One month after the vaccination, subjects with the AA and AG genotypes had a significantly higher T cell proliferative response against PT and FHA compared to those with the GG genotype. Subjects with the GG genotype had the lowest responses. As a conclusion, our preliminary results indicate that IL-10 SNP -1082 might play an important role in T cell-mediated immune responses after acellular pertussis vaccination.

A study on the effect of IL-6 gene polymorphism on the prognosis of non-small-cell lung cancer.

BackgroundLung cancer is one of the most commonly diagnosed clinical diseases. IL-6 is a multifunctional cytokine that is related to chemotactic factors and tumor biological regulation. −174G/C polymorphism in the promoter region of the IL-6 gene single-nucleotide polymorphism is the −174 position change from G to C. However, the relationship between the IL-6 gene polymorphism and prognosis of lung cancer is elusive. Therefore, the aim of this study was to evaluate the effect of −174G/C polymorphism on the prognosis of patients with non-small-cell lung cancer (NSCLC).MethodsDNA was extracted from the peripheral blood of 434 cases diagnosed with NSCLC by cytologic or histologic examination. Polymerase chain reaction–restriction fragment length polymorphism (NlaIII) was used to detect the genotype of −174G/C. Based on the functional activity of the IL-6 gene polymorphism, genotypes were divided into G vector (CG/GG) (high yield) and CC genotype (low yield). Prognosis of patients was analyzed and independent risk factors evaluated. A quantitative analysis of the degree of pain after diagnosis was performed to evaluate the correlations between gene polymorphisms and the degree of pain and use of analgesics.ResultsSurvival analysis showed that survival of the patients carrying the G allele (CG/GG) was significantly lower than that of patients with CC genotype (42.31 versus 62.79 months; P=0.032). The IL-6 gene promoter region revealed the presence of polymorphic variants, which may be associated with changes in the gene transcription process that affect the level of serum cytokines. IL-6 −174G/C gene polymorphism is associated with a significant morphine equivalent daily dose (IL-6 GG, 69.61; GC, 73.17; CC, 181.67; P=0.004). Homozygous IL-6 −174C/C genotype carriers required higher doses of opioids than GG or GC carriers.ConclusionPolymorphism of −174G/C in IL-6 is closely related to cancer pain in NSCLC patients, the use of analgesics, and survival prognosis. It is necessary to further confirm the related results and determine the underlying pathogenic mechanisms.

Molecular analysis of human population of Abbottabad Pakistan in relation to type-1 diabetes

Type 1 diabetes mellitus (T1DM) caused by the destruction of insulin producing beta cells of pancreas that are responsible for the autoimmune system of the individuals. Blood and urine glucose increases due to the lack of insulin in the body. The objective of the present study was to evaluate the role of IL-10 gene polymorphisms as susceptibility markers for Pakistani patients. IL-10 promoter polymorphisms (positions-592A/C) were analyzed in 183 individuals of Abbottabd. Ninety three known Pakistani patients with type-1 diabetes including 49 men and 44 women were recruited; having age ranged from 1.0 –70.0 years. Ninety one healthy individuals with no family history of diabetes were included as control. RFLP and PCR were used to detect allelic polymorphism of the IL-10 gene promoter region at position -592. Results indicated that the genotype frequency of the allel (592A) was associated with type-1 diabetes. The genotype frequency of -592A allele was higher in adult patients than in old age patients. Our investigation suggests that IL-10 gene polymorphisms could be involved in the development of type-1 diabetes in the Pakistani population.

Association of polymorphisms in IL6 gene promoter region with type 1 diabetes and increased albumin-to-creatinine ratio.

Pro-inflammatory cytokines, such as interleukin-6 (IL-6), have been considered as key factors in type 1 diabetes mellitus (T1DM) and diabetic nephropathy, thus, our aim was to investigate the association of IL6-174G>C (rs1800795) and -634C>G (rs1800796) polymorphisms with T1DM susceptibility and diabetic nephropathy. These polymorphisms were analyzed in 144 children and adolescents with T1DM and 173 normoglycemic control subjects. Glycemic control, laboratory parameters of kidney function and serum lipids were evaluated. By studying only T1DM patients, we evaluated the polymorphisms associated with relevant biochemical parameters in various genetic models. Type 1 diabetes mellitus patients showed poor glycemic control and albumin-to-creatinine ratio, total cholesterol and LDL-cholesterol levels increased when compared with normoglycemic subjects (p < 0.001, p = 0.004 and p < 0.001, respectively). IL6-174C allele was associated with an increased risk of developing T1DM (OR = 1.53, CI = 1.01-2.31, p = 0.044). In the T1DM group, IL6-174CC carriers showed higher concentrations of glycated hemoglobin (p = 0.029), albumin-to-creatinine ratio (p = 0.021), total cholesterol (p = 0.010), and LDL-cholesterol (p = 0.002), when compared with GG+GC carriers. No association was found for the IL6-634C>G polymorphism. These results suggest that IL6-174G>C may contribute to T1DM and increased albumin-to-creatinine ratio as well as to poor glycemic control and hyperlipidemia.

Influence of Production Affecting Cytokine Genetic Variants on Allogeneic Hematopoietic Cell Transplantation Outcomes

Introduction:High resolution human leukocyte antigen (HLA) matching is regarded as a prerequisite for the clinical success of allogeneic haematopoietic cell transplantation (HCT). However, studies have reported immunogenetic factors other than HLA also plays an important role in preventing and controlling post HCT complications, in particular graft-versus-host disease (GVHD) and infections. Cytokines and cytokine receptors, which are the key regulators of the immune surveillance against infections and/or allo-immune responses, are important candidates. The genetic control of cytokine production is evidenced by polymorphisms in cytokine gene regulatory regions resulting in low, moderate, or high cytokine production. Here, we evaluated the prognostic significance of cytokine genetic variants known to impact cytokine production on different outcomes of allogeneic HCT.Materials and Methods:A total of 793 subjects were accrued, including 360 adult allo-HCT donors (discovery cohort; n=240, and validation cohort; n=120), 382 HCT allo- recipients and 50 healthy individuals. All subjects of the discovery cohort and healthy individuals were analyzed for 22 single nucleotide variants located in the regulatory and/or exonic regions of 13 cytokine or cytokine receptor gene using sequence-specific primer based assay. All subjects of the validation cohort were genotyped SNPs located in the IL10 and IL1R gene promoter region by direct sequencing. . CMV specific immune response was assessed by stimulating PBMNCs from healthy individuals with CMV lysate and CMV peptide-pp65 followed by enumeration of IFN-g producing and CD107a expressing (degranulating) MNCs, T cells and their subsets.Results:Cytokine gene variants of donors and not that of recipients appear to influence post HCT complications. Two of these variant system- SNPs leading to low production of IL10 and high production of IL1R showed strong correlation with GVHD and posttransplant CMV infections respectively.Allo-HCT recipients who have received graft from donors carrying low IL-10 producing gene variants (AA) at IL-10 -1082G/A and (ATA/ATA at IL-10 -1082G/A, -819C/T and -592C/A) have high incidence of significant GVHD (defined as grade II-IV acute GVHD and/or chronic GVHD requiring systemic immunotherapy) (P=0.008, HR= 2.8, Figure 1; and P=0.03, HR= 1.8 respectively) compared to recipients who have received graft from donors carrying high producer IL-10 gene variants.Allo-HCT recipients who have received graft from donors carrying low IL-1R producing genotype (-1970 CC) showed high incidence of CMV reactivation (p=0.01; HR=2.1, Figure 2) in comparison to the HCT recipients receiving grafts from donors carrying high IL1-R producing genotype (-1970 GG). Further, the in-vitro culture analysis in healthy individuals showed that IL1R+ cells have 3-5 fold stronger anti-CMV immune responses (IFNg+ and/or CD107a+ cells) in comparison to IL1R- cells.Conclusions:Genetic predisposition to low IL-10 production is a strong predictor of GVHD while that tohigh production of IL-1R confers strong protection against CMV reactivation after allogeneic HCT. Thefindings implicate the importance of prospective assessment of cytokine gene variants to improve allogeneic HCT donor selection. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

A novel role for histone deacetylase 6 in the regulation of the tolerogenic STAT3/IL-10 pathway in APCs.

APCs are critical in T cell activation and in the induction of T cell tolerance. Epigenetic modifications of specific genes in the APC play a key role in this process, and among them histone deacetylases (HDACs) have emerged as key participants. HDAC6, one of the members of this family of enzymes, has been shown to be involved in regulation of inflammatory and immune responses. In this study, to our knowledge we show for the first time that genetic or pharmacologic disruption of HDAC6 in macrophages and dendritic cells results in diminished production of the immunosuppressive cytokine IL-10 and induction of inflammatory APCs that effectively activate Ag-specific naive T cells and restore the responsiveness of anergic CD4(+) T cells. Mechanistically, we have found that HDAC6 forms a previously unknown molecular complex with STAT3, association that was detected in both the cytoplasmic and nuclear compartments of the APC. By using HDAC6 recombinant mutants we identified the domain comprising amino acids 503-840 as being required for HDAC6 interaction with STAT3. Furthermore, by re-chromatin immunoprecipitation we confirmed that HDAC6 and STAT3 are both recruited to the same DNA sequence within the Il10 gene promoter. Of note, disruption of this complex by knocking down HDAC6 resulted in decreased STAT3 phosphorylation--but no changes in STAT3 acetylation--as well as diminished recruitment of STAT3 to the Il10 gene promoter region. The additional demonstration that a selective HDAC6 inhibitor disrupts this STAT3/IL-10 tolerogenic axis points to HDAC6 as a novel molecular target in APCs to overcome immune tolerance and tips the balance toward T cell immunity.

An initial assessment of correlations between host- and virus-related factors affecting analogues antiviral therapy in HBV chronically infected patients

BackgroundSuccess in treating hepatitis B virus (HBV) infection with nucleoside analogues drugs is limited by the emergence of drug-resistant viral strains upon prolonged therapy. In addition to mutation patterns in the viral polymerase gene, host factors are assumed to contribute to failure of treatment in chronic HBV infections. The aim of this study was to analyze the correlation between efficacy of antiviral therapy and the prevalence of HBV pretreatment drug-resistant variants. We also analyzed the role of heterogeneity in the promoter region of the IL-10 on the HBV pol/s gene polymorphisms and efficacy of analogues-driven therapy.Material/MethodsHBV DNA was extracted from 54 serum samples from chronic hepatitis B (CHB) patients. Drug-resistance mutations were analyzed using MALDI-TOF mass spectrometry technology (MALDI-TOF MS) and Multi-temperature single-strand conformation polymorphism (MSSCP). IL-10 gene promoter region polymorphisms at positions −1082, −819, and −592 were determined in allele-specific PCR reactions (AS-PCR).ResultsDrug-resistance mutations were detected in 74% of naïve and 93% of experienced patients, but the effect of pre-existence of drug-resistant HBV variants on antiviral therapy was not statistically significant (p=0.86). The role of polymorphisms at positions −1082 (p=0.88), −819 (p=0.26), and −592 (p=0.26) of IL-10 promoter region polymorphisms was excluded from the response-predicting factors. The main host factors predicting successful response to antiviral therapy were female sex (p=0.007) and young age (p=0.013).ConclusionsThe presence of drug-resistant HBV variants in baseline is not a viral predictor of good response to nucleoside/nucleotide analogues therapy. Only low HBV viral load predicted positive response to antiviral therapy. The ideal candidate for antiviral therapy is an immunocompetent, young female with low HBV viral load and elevated ALT activity.