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Single nucleotide polymorphisms (SNPs, rs12979860 e rs8099917) in the Interferon Lambda 4 gene (IFNL4, formerly IFNL3and/or IL28B) has been associated with failure in the innate immune response, sustained virological response in hepatitis C, and HTLV-1-associated myelopathy (HAM) development. To search for these polymorphisms several methodologies can be employed, such as sequencing, real-time or quantitative polymerase chain reaction (qPCR), restriction fragment length polymorphism analysis in PCR products (PCR-RFLP), and tetra-primer PCR. The present study compared the performance of the tetra-primer PCR in relation to the PCR-RFLP, both optimized in the Research HTLV Laboratory of the Center of Immunology of Instituto Adolfo Lutz in São Paulo. One hundred DNA samples obtained from patients of STD/Aids Reference Centre in São Paulo, previously analyzed for IL28B SNPs by PCR-RFLP were selected for analysis, after confirming that they represent all IL28B SNPs patterns described in the literature. The results obtained showed concordance between the PCR-RFLP and the tetra-primer PCR SNPs results, and because of the low cost, easy to perform, and minor employment of biological specimen and reagents, the tetra-primer PCR is of choice to be used in routine. (AU)
Polimorfismos de nucleotídeos únicos (single nucleotide polymorphisms, SNPs rs12979860 e rs8099917) no gene que codifica o Interferon Lambda 4 (IFNL4, antigamente IFNL3 e/ou IL28B) têm sido associados às falhas na resposta imune inata e resposta virológica sustentada na hepatite C, e a mielopatia associada ao HTLV-1 (HTLV-1-associated myelopathy, HAM). A pesquisa destes polimorfismos pode empregar diversas metodologias: sequenciamento, reação em cadeia da polimerase em tempo real ou quantitativa (quantitative polymerase chain reaction, qPCR), análise de fragmentos de restrição enzimática em produtos de PCR (restriction fragment length polymorphism in PCR products, PCR-RFLP) e a tetra-primer PCR. Este estudo comparou o desempenho da tetra-primer PCR em relação a PCR-RFLP, ambas otimizadas no Laboratório de Pesquisa em HTLV do Centro de Imunologia do Instituto Adolfo Lutz de São Paulo. Foram selecionadas 100 amostras de DNA obtidas de pacientes do Centro de Referência e Treinamento em DST/Aids de São Paulo cujos SNPs na IL28B foram anteriormente determinados por PCR-RFLP e representaram todos os perfis descritos em literatura. Os resultados obtidos mostraram concordância entre elas, e pelo fato da tetra-primer PCR ter menor custo, ser de fácil execução, empregar menos tempo, insumos e material biológico, é a técnica de escolha para uso em rotina. (AU)
Subject(s)
Polymorphism, Restriction Fragment Length , Polymerase Chain Reaction , Interleukins , Polymorphism, Single Nucleotide , Interferon LambdaABSTRACT
Objective To analyze the effect of single nucleotide polymorphism ( SNP) rs8099917 of interleukin-28B ( IL-28B) on spontaneous virus clearance and the efficacy of antiviral therapy in hepatitis C virus (HCV) infected patients in Huzhou area of Zhejiang Province.Methods A total of 268 HCV-infected patients were enrolled.The high sensitive HCV RNA quantification , HCV genotype and the IL-28B rs8099917 SNP were detected at baseline.One hundred and sixty-three patients received pegylated interferon α-2a ( Peg-IFNα-2a) and ribavirin (RBV) combined antiviral therapy (referred as PR treatment) for 48 weeks, who were followed up for 24 weeks.The remaining 44 patients were treated with sofosbuvir and daclatasvir for 12 weeks. The virological response of patients with different IL-28B rs8099917 genotypes was monitored.The count data was compared by χ2 test.Results The distributions of IL-28B rs8099917 SNP were 84.33%for TT genotype and 15.67%for GT genotype, while the GG genotype was not detected.Of the 135 patients with acute HCV infection, 61 cases had spontaneous viral clearance , 74 cases were converted to chronic infection.The spontaneous clearance rates were 47.11% for TT genotype and 28.57% for GT genotype.There was no significant difference of the spontaneous clearance rate between TT and GT genotype (χ2 ＝1.072, P＝0.30). In 163 chronic hepatitis C (CHC) patients with PR treatment, the rate of sustained virological response (SVR) after 24 weeks follow-up was 86.50%(141 cases).SVR rate in patients with TT genotype was significantly higher than those with GT genotype (91.67%vs 47.36%, χ2 ＝28.212, P＜0.05).There was no statistically significant difference of the SVR rates between genotype 1b and 2a (χ2 ＝1.525, P＞0.05).In 44 patients received sofosbuvir and daclatasvir treatment , both SVR rates of TT genotype and GT genotype were 100%. Conclusions In Huzhou area of Zhejiang Province , there is no significant correlation between IL-28B rs8099917 genotype and spontaneous clearance in patients with acute HCV infection , but the genotype of IL-28B rs8099917 is valuable for the prediction of PR treatment efficacy , the SVR rate of the TT genotype is superior to the GT genotype.The SVR rate can reach 100% in patients received combination therapy of sofosbuvir and daclatasvir independent of polymorphism of IL-28B.
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Background & objectives: The effect of vitamin D supplementation on response to antiviral therapy in hepatitis C virus (HCV) genotype 1 and 4 infection still remains unclear, with studies yielding inconsistent results. The aim of the present study was to assess the effect of vitamin D supplementation on treatment outcome in patients with genotype 1/4 chronic hepatitis C (CHC) infection. Methods: Sixty consecutive, treatment-naïve, genotype 1 and 4 chronic HCV patients were included in the study. The patients were randomized into two groups: Vitamin D supplemented group received pegylated (PEG)-interferon ?-2a 180 ?g per week plus ribavirin (RBV) (1000-1200 mg/d) together with vitamin D3 (2000 IU/d) and control group received identical therapy without vitamin D (32 patients). Results: There were no significant differences between the two groups in terms of age, sex, body mass index and baseline laboratory values. Lower vitamin D levels were associated with higher grades of fibrosis in liver histology (vitamin D >20 ng/ml - 70% vs vitamin D <20 ng/ml - 37%, P<0.05). Vitamin D supplemented group had similar rapid viral response (40 vs 28%, P=0.36), complete early viral response (53.2 vs 40%, P=0.34), end of treatment response (64 vs 46%, P=0.17) and sustained virological response (SVR) (60 vs 44%, P=0.19) as compared to control group. Interleukin 28B polymorphism [odds ratio (OR)-15.37, 95% confidence interval (CI)-2.32-101.76, P=0.04] and baseline serum vitamin D levels (OR-6.36, 95% CI-1.36-29.61 P=0.02) were independent predictors of SVR in genotype 1/4 CHC. Vitamin D supplementation was not found to be predictor of response in genotype 1/4 CHC on multivariate analysis (OR-2.79, 95% CI- 0.63-12.34, P=0.74). Interpretation & conclusions: The present study showed that addition of vitamin D to PEG/RBV combination therapy in treatment-naïve patients who were infected with HCV genotype 1/4 had no effect on the rates of rapid, early and sustained viral responses.
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BACKGROUND/AIMS: The present study aimed to evaluate the safety and efficacy of simeprevir-based triple therapy with reduced doses of pegylated interferon (PEG-IFN) and ribavirin for interferon (IFN) ineligible patients, such as elderly and/or cirrhotic patients, and to elucidate the factors contributing to a sustained virologic response (SVR). METHODS: One hundred IFN ineligible patients infected with genotype 1b hepatitis C virus (HCV) were treated. Simeprevir (100 mg) was given orally together with reduced doses of PEG-IFN-α 2a (90 μg), and ribavirin (200 mg less than the recommended dose). RESULTS: The patients’ median age was 70 years, and 70 patients were cirrhotic. Three patients (3%) discontinued treatment due to adverse events. The SVR rate was 64%. Factors that significantly contributed to the SVR included the γ-glutamyl transferase and α-fetoprotein levels, interleukin-28B (IL28B) polymorphism status, and the level and reduction of HCV RNA at weeks 2 and 4. The multivariate analysis showed that the IL28B polymorphism status was the only independent factor that predicted the SVR, with a positive predictive value of 77%. CONCLUSIONS: Simeprevir-based triple therapy with reduced doses of PEG-IFN and ribavirin was safe and effective for IFN ineligible patients infected with genotype 1b HCV. IL28B polymorphism status was a useful predictor of the SVR.
Subject(s)
Aged , Humans , Genotype , Hepacivirus , Hepatitis C , Hepatitis , Interferons , Multivariate Analysis , Ribavirin , RNA , Simeprevir , TransferasesABSTRACT
Objective To investigate the relationship between interleukin (IL)-28B gene polymorphisms (rs12979860 and rs8099917) and treatment response in patients with chronic hepatitis C in China.Methods Taqman probes single nucleotide polymorphism genotyping methods were used to detect the genotypes of rs12979860 (C/T) and rs8099917 (T/G) located at IL-28B gene in 105 included patients.The patients were treated with standard doses of pegylated interferon plus ribavirin and were followed up regularly for therapeutic response and adverse reaction.The relationship between IL-28B gene polymorphism and antiviral treatment response of patients were analyzed.Categorical data were analyzed using Pearson chi-square test or Fisher exact test.Results Totally 105 cases were included in our study and 2 cases lost to follow-up because of moving away.Eight-one cases (78.6%) of the remaining 103 patients were CC/TT genotype (CC/TT group) at rs12979860 and rs8099917, 19 cases (18.4%) were CT/TG (CT/TG group) and 3 cases (2.9%)were TT/TG (TT/TG group).No other genotypes were detected and linkage disequilibrium was discovered at the two polymorphism loci (r2=0.11).After 4 weeks of treatment, 35 cases (43.2%) in CC/TT group, 3 cases (15.8%) in CT/TG group and non in TT/TG group achieved rapid virological response (RVR).There were statistically significant differences among three groups (P=0.033).After 12 weeks of treatment, 45 cases (55.6%) in CC/TT group, 6 cases (31.6%) in CT/TG group and none in TT/TG group achieved early virological response (EVR).There were statistically significant differences among three groups (P=0.025).At the end of the treatment, 68 cases (83.9%) in CC/TT group, 10 cases (52.6%) in CT/TG group and only 1 case (33.3%) in TT/TG group achieved end-of-treatment response (ETR).There were significant statistical differences among the three groups (P=0.003).After 24 weeks of follow-up, 62 cases (76.5%) in CC/TT group, 9 cases (47.4%) in CT/TG group and 1 case (33.3%) in TT/TG group achieved sustained virological response (SVR).There were statistically significant differences among the three groups (P=0.014).One hundred and one cases in CC/TT group developed adverse events, among them 19 cases needed clinical treatment.There were 43 cases in CT/TG group developed adverse events and 9 cases needed treatment.Seven cases in TT/TG group developed adverse events and only 1 case needed treatment.There were no statistically significant difference among three groups (χ2=0.139,P>0.05).Conclusions The genotype of rs12979860 (C/T) and rs8099917 (T/G) at IL-28B gene could affect the treatment response in patients with chronic hepatitis C.RVR and SVR are higher in patients with genotype CC/TT, which might help to guide HCV treatment.
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Objective To explore the association between interleukin 28B (IL-28B),equilibrative nucleoside transporters 1 (ENT1) gene polymorphisms and spontaneous clearance of HCV in HIV/HCV co-infectors in Hunan province.Methods Genotypes of IL-28B and ENT1 (rs12980275,rs12979860,rs8099917 and rs760370) were analyzed in 107 HIV/HCV co-infectors in Hunan province and the distributions of gene polymorphisms were compared between chronic hepatitis and spontaneous clearance groups.Results The major genotypes in rs12980275,rs12979860 and rs8099917 of IL-28B were AA,CC and TT in HIV/HCV co-infectors,which accounted for 84.1％ of each.The three single nucleotide polymorphisms were highly linkage disequilibrium (r2＞0.94) and the differences of genotype distribution were statistically significant between chronic hepatitis and the spontaneous clearance groups (P＜0.05).Infectors which carrying the major genotypes were more susceptible to spontaneous clearance of HCV.Differences of the genotype distributions in rs760370 of ENT1 were not statistically significant between the two groups.Conclusion Genotypes AA,CC and TT of IL-28B were related to spontaneous clearance of HCV in HIV/HCV co-infectors.
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Objective To explore the correlation between polymorphism of interleukin-28B(IL-28B) rs12979860 T/ C and susceptibility of hepatocellular carcinoma( HCC). Methods All eligible case-control studies published up to 2014-09-30 were identified by searching PubMed,EMBase,CNKI,CBM,VIP and WanFang databases. Two reviews independently identified the literature according to inclusion and exclusion criteria. Meta-analysis was performed using Rev Man 5. 2 and Stata 12. 0 software. Results A total of 6 stud-ies comprising 1 138 cases and 955 controls were finally included. Meta-analysis showed that IL-28B rs12979860 T/ C polymorphism was associated with the susceptibility of HCC. Compared with the genotype CC and CT + CC,genotype TT increased the risk of suffering from HCC(TT vs CC:OR = 2. 26,95% CI:1. 40-3. 64,Z = 3. 33,P = 0. 000 9;TT vs CT + CC:OR = 1. 90,95% CI:1. 23-2. 93,Z = 2. 89,P = 0. 004). In stratification analysis by ethnicity,we observed that the polymorphism of IL-28B rs12979860 T/ C was associat-ed with the susceptibility of HCC among Caucasian populations(TT vs CC:OR = 2. 06,95% CI:1. 22-3. 47, Z = 2. 70,P = 0. 007;TT vs CT + CC:OR = 1. 71,95% CI:1. 07-2. 72,Z = 2. 23,P = 0. 03). Conclusion The polymorphism of IL-28B rs12979860 T/ C is associated with the susceptibility of HCC,genotype TT may increase the susceptibility to HCC.
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PURPOSE: The role of IL28B gene variants and expression in hepatitis B virus (HBV) infections are not well understood. Here, we evaluated whether IL28B gene expression and rs12979860 variations are associated with HBV outcomes. MATERIALS AND METHODS: IL28B genetic variations (rs12979860) were genotyped by pyrosequencing of DNA samples from 137 individuals with chronic HBV infection [50 inactive carriers (IC), 34 chronic hepatitis B (CHB), 27 cirrhosis, 26 hepatocellular carcinoma (HCC)], and 19 healthy controls. IL28A/B mRNA expression in peripheral blood mononuclear cells was determined by qRT-PCR, and serum IL28B protein was measured by ELISA. RESULTS: Patients with IL28B C/C genotype had greater IL28A/B mRNA expression and higher IL28B protein levels than C/T patients. Within the various disease stages, compared to IC and healthy controls, IL28B expression was reduced in the CHB, cirrhosis, and HCC cohorts (CHB vs. IC, p=0.02; cirrhosis vs. IC, p=0.01; HCC vs. IC, p=0.001; CHB vs. controls, p<0.01; cirrhosis vs. controls, p<0.01; HCC vs. controls, p<0.01). When stratified with respect to serum HBV markers in the IC and CHB cohorts, IL28B mRNA and protein levels were higher in HBeAg-positive than negative individuals (p=0.01). Logistic regression analysis revealed that factors associated with high IL28B protein levels were C/C versus C/T genotype [p=0.016, odds ratio (OR)=0.25, 95% confidence interval (CI)=0.08-0.78], high alanine aminotransferase values (p<0.001, OR=8.02, 95% CI=2.64-24.4), and the IC stage of HBV infection (p<0.001). CONCLUSION: Our data suggest that IL28B genetic variations may play an important role in long-term development of disease in chronic HBV infections.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Alanine Transaminase/blood , Asian People/genetics , Biomarkers/blood , Carcinoma, Hepatocellular/genetics , Case-Control Studies , China , DNA, Viral/blood , Enzyme-Linked Immunosorbent Assay , Genotype , Hepatitis B virus/genetics , Hepatitis B, Chronic/ethnology , Interleukins/blood , Leukocytes, Mononuclear , Liver Cirrhosis/blood , Liver Neoplasms/genetics , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
BACKGROUND/AIMS: There are few available data regarding the association between the single nucleotide polymorphisms (SNPs) of the gene encoding interleukin 28B (IL28B) and a sustained virologic response (SVR) to peginterferon (PEG-IFN) plus ribavirin (RBV) therapy in Korean chronic hepatitis C patients. METHODS: This was a retrospective cohort study of 156 patients with chronic hepatitis C virus (HCV) infection who received combination treatment of PEG-IFN plus RBV. Blood samples from these patients were analyzed to identify the IL28B SNPs at rs12979860, rs12980275, rs8099917, and rs8103142. Association analyses were performed to evaluate the relationships between each IL28B SNP and SVRs. RESULTS: Seventy six patients with HCV genotype 1 and 80 with genotype non-1 were enrolled. The frequencies of rs12979860 CC and CT genotypes were 90.4% and 9.6%, respectively; those of rs12980275 AA and AG genotypes were 87.2% and 12.8%, respectively; those of rs8099917 TT and TG genotypes were 92.3% and 7.7%, respectively; and those of rs8103142 TT and CT genotypes were 90.4% and 9.6%, respectively. Among the patients with HCV genotype 1, the SVR rates were 69.7% and 80.0% for rs12979860 CC and CT, respectively (P=0.71). Among the HCV genotype non-1 patients, SVR rates were 88.0% and 100% for rs12979860 CC and CT (P=1.00), respectively. CONCLUSIONS: Genotypes of the IL28B SNP that are known to be favorable were present in most of the Korean patients with chronic hepatitis C in this study. Moreover, the IL28B SNP did not influence the SVR rate in either the HCV genotype 1 or non-1 patients. Therefore, IL28B SNP analysis might be not useful for the initial assessment, prediction of treatment outcomes, or treatment decision-making of Korean chronic hepatitis C patients.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Alleles , Antiviral Agents/therapeutic use , Asian People/genetics , Cohort Studies , Drug Therapy, Combination , Gene Frequency , Genotype , Hepatitis C, Chronic/drug therapy , Interferon-alpha/therapeutic use , Interleukins/genetics , Linkage Disequilibrium , Polyethylene Glycols/therapeutic use , Polymorphism, Single Nucleotide , Recombinant Proteins/therapeutic use , Republic of Korea , Retrospective Studies , Ribavirin/therapeutic useABSTRACT
BACKGROUND/AIMS: This study investigated the role of single nucleotide polymorphisms (SNPs) near the interleukin-28B (IL28B) gene with respect to clinical outcomes and the antiviral response in hepatitis C virus (HCV) infection to suggest the practical utility of IL28B genotyping in Korea. METHODS: Two SNPs near IL28B, rs12979860 and rs8099917, were analyzed using an allelic discrimination assay in a total of 454 individuals, including 147 health-check examinees and 307 patients with HCV infection. RESULTS: The CC genotype frequency was significantly higher in the spontaneous recovery group than in the chronic infection group and was higher in the chronic hepatitis group than in the liver cirrhosis or hepatocellular carcinoma group, suggesting its favorable role in the clinical outcome. Multivariate analysis revealed that the rs12979860 CC genotype was an independent predictor of sustained virologic response (SVR) in genotype 1 HCV infection. During the currently used response-guided therapy, IL28B genotyping was most helpful for the patients who exhibit early virologic responses without rapid virologic responses, as those patients exhibiting the non-CC type did not achieve SVR, although they represented approximately one-third of the total patients. CONCLUSIONS: The IL28B SNP is an independent predictor of SVR. Our results may be helpful if the findings are carefully applied to select patients in Korea.
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular/genetics , Hepatitis C, Chronic/genetics , Interleukins/genetics , Liver Cirrhosis/genetics , Liver Neoplasms/genetics , Polymorphism, Single Nucleotide , Prognosis , Republic of Korea , Retrospective StudiesABSTRACT
Objective To investigate the correlation between IL-28B rs8099917 polymorphism and the outcome of HBV infection.Methods Genotype ofrs8099917(T＞G) in IL-28B locus was determined by TaqMan SNP genotyping from 486 individuals which including 199 chronic HBV carriers (including 100 HBV-induced liver cirrhosis and 99 HBV-related HCC).143 people with selflimited infection and 144 healthy people served as controls.Multivariate analysis was used to assess the effect of IL-28B rs80999 1 7 SNP among all the studied groups.Results Distribution of genotype and allele of the rs8099917 locus were in accordance with Hardy-Weinberg equilibrium in different groups or with the total population.The frequencies of the rs8099917 TT,GT,GG genotypes were 89.3％,10.5％ and 0.2％,and the frequency of allele T and G accounted for 94.5％ and 5.5％,respectively.In respect of genotype or allele frequency,there was no significant differences found among the groups(P＞0.05 ).When comparing with the TT genotype,data from the multinomial logistic analysis showed that the ORs and (95％CI) of TG/GG genotypes were 1.589 (0.735-3.437),1.351 (0.550-3.316) and 1.704 (0.717-4.052),respectively.The genotype frequencies in different groups with different clinical features showed that TG/GG genotypes significantly increased the risk of r-GT Ⅱ( + ) for individuals with HBV-related HCC (X2=17.534,P=0.001 ),with OR as 14.821 (3.227-68.064).It was particularly so for males(X2=14.924,P=0.014),with OR(95％CI) as 45.000(2.772-730.571 ).Conclusion IL-28B rs8099917 SNP had no correlation with the outcome of HBV infection.