- Research article
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Polymorphisms in the Th17 cell-related RORC gene are associated with spontaneous clearance of HCV in Chinese women
BMC Infectious Diseases volume 18, Article number: 254 (2018)
Abstract
Background
Female gender and favorable IFNL3 genotypes are the primary independent predictors of spontaneous clearance of HCV infection. However, chronic hepatitis C infection occurs in numerous women carrying favorable IFNL3 genotypes, indicating that other host and/or virological factors contribute to the prognosis of infection.
Methods
A cohort of 137 anti-HCV-positive female Han Chinese cases, including 64 chronic HCV carriers and 73 HCV spontaneous resolvers, was recruited in the study. 111 SNPs in 23 genes encoding HCV co-receptors, transcription factors, Toll-like receptors, co-stimulating molecules, and cytokines were selected for SNP analysis.
Results
After comparison of genotypes and allelotype frequencies of 111 SNPs in 23 genes in the primary cohort, the SNPs rs9826 (P = 0.024 for CC/TT/CT; P = 0.015 for C allele/T allele) and rs1521177 (P = 0.017 for GG/TT/GT; P = 0.006 for G allele/T allele) in the RORC gene were significantly associated with spontaneous HCV clearance. In the sub-cohort carrying favorable IFNL3 genotypes (rs12979860CC, rs8099917 TT, rs12980275 AA), rs1521177 (genotype: P = 0.040; allelotype: P = 0.021) remained significantly associated with spontaneous HCV clearance. Importantly, the most common RORC haplotype rs9826-T/rs1521177-T was presented at significantly different frequencies in resolvers and carriers in both the primary cohort (P = 0.0027) and the IFNL3 favorable sub-cohort (P = 0.0117).
Conclusions
This study indicates that genetic polymorphisms in human Th17-related RORC gene are associated with different natural prognosis of HCV infection. The RORC haplotype, rs9826-T/rs1521177-T, was favorable for spontaneous clearance of HCV infection.
Background
Chronic hepatitis C infection is a global healthcare burden and associated with the development of liver cirrhosis, hepatocellular carcinoma, and death if untreated [1]. In China, the prevalence of HCV in the general population is estimated as 1.6% [2, 3], which equals to approximately 20 million HCV carriers. A minority of acutely infected individuals (approximately 15–40%) will resolve their infection and recover spontaneously [4]. It is reported that among host factors, gender, age, IFNL3 (previously IL28B) genotypes, KIR/HLA alleles, ethnicity, and HIV co-infection are associated with spontaneous recovery [5,6,7,8,9,10,11,12,13,14,15]. In particular, female sex and favorable IFNL3 genotypes (rs12979860 CC, rs8099917 TT and rs12980275 AA) are primary independent predictors of spontaneous clearance [5,6,7,8,9, 12, 13, 16, 17]. In the general Han Chinese ethnic population, the three favorable IFNL3 genotypes are very common (~ 90%) [12, 16,17,18]. However, many females carrying favorable IFNL3 genotypes suffer from chronic hepatitis C in China, indicating that besides IFNL3 genotypes and possibly some KIR/HLA alleles, other immune-related factors associated with HCV spontaneous clearance still remain unknown.
T helper 17 cells (Th17), a subset of pro-inflammatory T helper cells, play an important role in adaptive immunity, protecting hosts against pathogens primarily through secreting cytokines(such as IL-17A, IL-17F, IL-21, and IL-22) [19]. Receptor retinoic acid-related orphan receptor C (RORC), a DNA-binding transcription factor, is the master regulator of Th17 differentiation [20, 21]. Th17 cells may have dual roles (both harmful and beneficial) in HCV-related disease [22]. Specific HCV-Th17 cells are involved in immune response modulation, and their levels are associated with severity of fibrosis and intrahepatic inflammatory status [23, 24]. On the other hand, several reports indicate that elevated Th17 responses could be associated with spontaneous HCV clearance [25, 26].
In this study, two single nucleotide polymorphisms (SNPs) located in the RORC gene were identified to be associated with spontaneous clearance of HCV infection.
Methods
Participants
In the present study, 190 HBsAg-negative, anti-HCV-positive, Han Chinese females were identified by screening a total of 1252 residents (80% of the local population) in Wangying Village, Shangcai County, Henan province, in August 2009. More than 90% of patients were former plasma donors, and the remaining individuals composed of their parents, spouses, or children. Samples were tested for anti-HCV antibody using the Architech anti-HCV system (Abbott Diagnostics, USA), and those with signal/cut-off ratios between 1.0 and 5.0 were confirmed by RIBA assay (HCV BLOT 3.0, MP diagnostics, USA). Plasma HCV viral load was determined using the Abbott RealTime HCV Amplification Kit (Abbott Molecular Inc., USA) according to the manufacturer’s instructions. Spontaneous resolvers were defined as subjects who were positive for HCV antibody and negative for plasma HCV RNA with no history of HCV-specific treatment. HCV RNA-negative status was confirmed in a second sample collected in 2012 and/or 2013. Chronic HCV carriers were defined as those patients positive for both HCV antibody and plasma HCV viral load and also confirmed by the follow-up detection. No participants had received any type of HCV-specific antiviral therapy. All subjects were screened for HIV-1 infection status. Anti-HIV-1 antibody status was initially tested by ELISA assay (GBI biotech Co., Ltd., Beijing, China) and confirmed by HIV Blot 2.2 WB assay (Genelabs Diagnostics, Singapore). All HIV-positive patients had received first-line antiretroviral therapy for 6~ 8 years, regularly or intermittently. None of the participants received any types of anti-HCV treatment.
Of 190 female anti-HCV-positive individuals, HIVpos women with chronic HCV infection (n = 53) were excluded from the study, as HIV infection could affect the ability of some individuals to spontaneously resolve their HCV infection. The final case number of primary cohort was 137, which consisted of a “Chronic” group (HIVneg HCV carriers, n = 64) and a “Resolved” group (n = 73, including 45 HIVneg HCV resolvers and 28 HIVpos HCV resolvers). Overall, the cohort exhibited a similar high frequency of IFNL3 favorable genotypes to Han Chinese nationality (Additional file 1: Table S1), indicating that the study was based on a representative cohort of the Chinese population [12, 16]. To eliminate the influence of IFNL3 SNPs on viral clearance, a sub-cohort carrying favorable IFNL3 genotypes (rs12979860CC, rs8099917TT, rs12980275 AA) was constructed and analyzed. The IFNL3 favorable sub-cohort included 54 and 67 participants in the “Chronic” and “Resolved” (HIVneg, n = 43; HIVpos, n = 24) groups, respectively. A flow diagram for cohort construction was provided as Fig 1. The clinical and biochemical characteristics of the primary cohort were presented in Table 1.
SNP selection and genotyping
A total of 111 SNPs in 23 genes encoding HCV co-receptors (CD81, SCARB1, CLDN1, OCLN, NPC1L1, APOE, LDLR), transcription factors(IRF3, RORC, TBX21, FOXP3, BCL6), Toll-like receptors (TLR3, TLR7, TLR9), co-stimulating molecules (ICOS, CXCR5, CD40LG), and cytokines(IL4, IFNG, IL21, IFNL3, CXCL13) were selected for SNP analysis (Additional file 1: Table S2). The selected SNPs met at least one of the following criteria:1) a reported minor allele frequency > 10% in the Han Chinese population according to SNP browser software 4.0 (Applied Biosystems), with reference to the NCBI SNP database (http://www.ncbi.nlm.nih.gov/projects/SNP/); 2) published evidence that the SNP was associated with disease.
Genomic DNA extracted from whole blood samples using a DNeasy Blood & Tissue kit (Qiagen, USA) was dissolved in sterile double distilled water and stored at − 20 °C until use. DNA purity was checked and DNA samples with 260/280 ratios< 1.7 were re-purified. SNP genotyping was performed using the iPLEX Sequenom MassARRAY system (Sequenom Inc., USA).
Statistical analyses
Statistical and graphical analyses were performed using GraphPad Prism 5.0, Microsoft Excel 2007, or SPSS 20.0. The allele frequency and genotype distributions of each SNP were descriptively summarized as numbers of cases and frequencies. Chi-square (χ2) and Fisher’s exact tests were used to examine differences in frequencies of individual SNPs between HCV carriers and spontaneous resolvers. SNP-specific deviation from Hardy–Weinberg Equilibrium (HWE) in the whole study population was tested using a χ2 test in SHEsis software, and SNPs with HWE violation (P < 0.01) were excluded. Pairwise D′ and r2 measures of linkage disequilibrium for RORC SNPs were calculated using SHEsis (http://analysis.bio-x.cn/myAnalysis.php). Specific parameters were set as previously described [27]. P-values, odds ratios (ORs), and 95% confidence intervals (95% CIs) were used for association analysis. All P-values were two-tailed, and were considered significant at < 0.05.
Results
Associations between target gene SNPs and spontaneous HCV clearance in the primary cohort
Genotype distributions and allele frequencies were calculated for 111 candidate SNPs in both the “Chronic” and “Resolved” groups of the primary cohort (Additional file 1: Table S2). The frequencies of favorable IFNL3 genotypes were higher in “Resolved” group than “Chronic” group (resolved vs. chronic: rs12979860, CC 96% vs. 86%; rs8099917, TT 95% vs. 91%; and rs12980275, AA 96% vs. 86%), although only the difference in the frequency of the rs12979860 C allele was statistically significant (P = 0.043) (Table 2).
Of the 22 remaining genes tested (excluding IFNL3), only SNPs rs9826 and rs1521177, located in non-coding regions of RORC, exhibited frequency differences between the “Resolved” and “Chronic” groups (rs9826: P = 0.024 for CC/TT/CT, P = 0.015 for C allele/T allele, OR = 1.97, 95% CI: 1.14-3.41; rs1521177: P = 0.017 for GG/TT/GT, P = 0.006 for G allele/T allele, OR = 2.18, 95% CI: 1.24-3.82)(Table 2). The rs9826 TT and rs1521177 TT genotypes were associated with improved viral clearance. Overall, the primary cohort exhibited a similar distribution of RORC genotypes to Han Chinese nationality (Additional file 1: Table S3). The Hardy–Weinberg Equilibrium (HWE) test of three IFNL3 SNPs and two RORC SNPs in the primary cohort were tested, and no SNPs with HWE violation were presented in the study (Additional file 1: Table S4).
In the present study, a minority of “Resolved” individuals were co-infected with HIV (HIVpos resolved, n = 28) and very similar IFNL3 (rs12979860, rs8099917, and rs12980275) and RORC (rs9826 and rs1521177) SNP genotype distributions were identified among HIVneg, HIVpos, and total resolved individuals (Additional file 1: Figure S1). No association was observed between RORC SNPs (rs9826 and rs1521177) and HCV RNA levels in HIV-uninfected “Chronic” individuals (Additional file 1: Figure S2). In addition, we did not find any differences in genotypes and allele frequency distributions of RORC SNPs (rs9826 and rs1521177) between HIVpos HCV carriers and HIVpos HCV resolvers (Additional file 1: Table S5), and no associations between RORC SNPs and type of HCV genotypes (HCV 2a vs. HCV 1b) was found in HIVneg HCV carriers (Additional file 1: Table S6).
Re-evaluation of RORC SNP associations with spontaneous HCV clearance in the IFNL3 favorable sub-cohort
In the IFNL3 favorable sub-cohort, cases with unfavorable IFNL3 genotypes in the “Chronic” (n = 10) and “Resolved” (n = 6) groups were excluded. Analysis of the sub-cohort indicated that significant differences between the two groups remained for rs1521177 (P = 0.040 for GG/TT/GT, P = 0.021 for G allele/T allele, OR = 2.00, 95% CI: 1.11-3.55), although no significant differences were identified for rs9826 (P = 0.092 for CC/TT/CT, P = 0.062 for C allele/T allele, OR = 1.74, 95% CI: 0.97-3.19) (Table 3). These data suggested that SNP alleles in RORC had an independent effect on HCV viral clearance, which couldnot be explained by the distribution of IFNL3 polymorphisms.
Analysis of the RORCrs9826/rs1521177 haplotype
Linkage disequilibrium tests for RORC and IFNL3 SNPs in the primary cohort were shown in Additional file 1: Figure S3. The RORC SNPs, rs9826 and rs1521177, were determined to be in linkage disequilibrium (Primary cohort: D’ = 0.831, r2 = 0.639; IFNL3 favorable sub-cohort: D’ = 0.812, r2 = 0.645) (Additional file 1: Table S7). Two main haplotypes were present in the primary cohort (rs9826-T/rs1521177-T: 62% vs. 79%, chronic vs. resolved; rs9826-C/rs1521177-G: 27% vs. 16%, chronic vs. resolved) and the IFNL3 favorable sub-cohort (rs9826-T/rs1521177-T: 64% vs. 78%, chronic vs. resolved; rs9826-C/rs1521177-G: 26% vs. 17%, chronic vs. resolved). Overall, the presence of the rs9826-T/rs1521177-T haplotype was associated with a higher likelihood of spontaneous resolution in both the primary cohort (P = 0.0027, OR = 0.445, 95% CI: 0.26–0.76) and the IFNL3 favorable sub-cohort (P = 0.0117, OR = 0.485, 95% CI:0.28–0.86). In addition, the frequency of the rs9826-C/rs1521177-G haplotype was significantly different between the chronic and resolved groups in the primary cohort (P = 0.0315, OR = 1.893, 95% CI:1.05–3.40), suggesting that individuals carrying this haplotype was difficult in spontaneous resolution of HCV infection (Table 4).
Discussion
Genetic polymorphisms of innate immunity-related genes were reported to be associated with HCV clearance [28, 29]. Specially, SNPs of the IFNL3 (IL-28B) have shown strong association with spontaneous clearance of HCV and with the response to anti-HCV therapy [30,31,32,33]. Another important correlation of HCV spontaneously viral clearance was a robust and sustained HCV-specific T cell response. Cumulative studies indicated that HLA-associated viral polymorphisms were closely correlated with immune recognition of T cells to virus (both HIV and HCV) [34,35,36,37].
Exploration of adaptive immune factors involved in spontaneous clearance of viral infection will help to understand HCV pathogenesis. Besides IFNL genotypes, HCV-specific T cells are typically detectable 5–9 weeks after acute infection [38, 39]. Recently, Grebely et al. reported that the median time for acute HCV clearance among 173 spontaneous resolvers undergoing follow-up observation was 16.5 weeks [9], indicating that a strong, broad, and persistent HCV-specific adaptive immune response is required during acute infection for viral clearance [40].
The role of Th17 responses in HCV pathogenesis is intriguing but not well characterized. Numerous studies have focused on its destructive potentials, since they could aggravate the severity of fibrosis and intrahepatic inflammatory status [22, 41, 42]. However, some studies have also reported a role for Th17 responses in spontaneous HCV clearance. Kared et al. reported that the differentiation of IL-17A-producing CD4+ T cells was correlated with prognosis of infection [26]. In addition, the concentration of plasma IL-17A was significantly higher in the acute phase of HCV infection in patients with self-limiting infection than in those with chronically evolving hepatitis [26]. Moreover, Seetharam et al. described a transient IL-17 response followed by a subsequent reactivation of Th1 responses, resulting to spontaneous recovery in a liver transplant recipient with recurrent hepatitis C [25]. It is possible that Th17 responses in HCV-associated disorders may act as a double-edged sword, playing different roles in different disease courses.
In this study, a number of the HCV resolvers had a background of HIV infection, which could be primarily ascribed to historical contaminated commercial blood donations in the late 1990s. As HCV is more efficiently transmitted through blood contamination or infected needles than HIV, self-limiting acute HCV infection in these individuals may usually precede, or be coincident with, HIV infection [6, 43, 44]. Regardless of whether spontaneous recovery from HCV occurred before or after HIV infection, it remains conceivable that genetic immune characteristics of these individuals could be associated with spontaneous HCV eradication. By contrast, we excluded chronically HIV/HCV-co-infected patients from the study, due to the possibility that some HIV-co-infected individuals who could clear HCV spontaneously in the absence of HIV became chronic HCV carriers. In addition, all the participants in this study came from the same village and shared similar characteristics of viral contamination mode, daily diet, surrounding environment, ethnicity, and income level, which strengthed the comparison of genetic immune factors between HCV carriers and resolvers. Among the two RORC SNPs (rs9826 and rs1521177) identified in this study, the rs9826 polymorphism was associated with severity of rheumatoid arthritis in the Polish population [45]. Further functional studies to explore the associations between RORC SNPs genotypes and Th17 responses, as well as Th17 responses and viral clearance, should be performed in the future.
Our cohort design precluded the influence factors of ethnicity, gender and IFNL-3. However, the sample size of our cohort was still limited, and future larger-scale queues were needed for better verification of our conclusion. In addition, the RORC SNPs rs9826-T and rs1521177-T were validated in only Chinese Han population in this study. The adaptation of these SNPs in other ethnic and national populations needs to be further analyzed.
Conclusions
The present study determined that polymorphisms in human Th17-related RORC gene were associated with differing natural prognosis for Chinese Han females with HCV infection. The most common RORC haplotype in this cohort, rs9826-T/rs1521177-T, was favorable for spontaneous clearance of HCV infection.
Abbreviations
- 95% CIs:
-
95% confidence intervals
- HAART:
-
High active antiretroviral therapy
- HCC:
-
Hepatocellular carcinoma
- HCV:
-
Hepatitis C virus
- HIV:
-
Human immunodeficiency virus
- IFNL3 :
-
Interferon lambda 3
- LD:
-
Linkage disequilibrium
- OR:
-
Odds ratios
- RORC:
-
Receptor retinoic acid-related orphan receptor C
- SNP:
-
Single nucleotide polymorphisms
- Th17:
-
T helper 17 cells
References
Messina JP, Humphreys I, Flaxman A, Brown A, Cooke GS, Pybus OG, Barnes E. Global distribution and prevalence of hepatitis C virus genotypes. Hepatology. 2015;61(1):77–87.
Kao JH, Ahn SH, Chien RN, Cho M, Chuang WL, Jeong SH, Liu CH, Paik SW. Urgency to treat patients with chronic hepatitis C in Asia. J Gastroenterol Hepatol. 2017;32(5):966–74.
Bennett H, Waser N, Johnston K, Kao JH, Lim YS, Duan ZP, Lee YJ, Wei L, Chen CJ, Sievert W, et al. A review of the burden of hepatitis C virus infection in China, Japan South Korea and Taiwan. Hepatol Int. 2015;9(3):378–90.
Mohd Hanafiah K, Groeger J, Flaxman AD, Wiersma ST. Global epidemiology of hepatitis C virus infection: new estimates of age-specific antibody to HCV seroprevalence. Hepatology. 2013;57(4):1333–42.
Huang J, Huang K, Xu R, Wang M, Liao Q, Xiong H, Li C, Tang X, Shan Z, Zhang M, et al. The associations of HLA-A*02:01 and DRB1*11:01 with hepatitis C virus spontaneous clearance are independent of IL28B in the Chinese population. Sci Rep. 2016;6:31485.
van den Berg CH, Grady BP, Schinkel J, van de Laar T, Molenkamp R, van Houdt R, Coutinho RA, van Baarle D, Prins M. Female sex and IL28B, a synergism for spontaneous viral clearance in hepatitis C virus (HCV) seroconverters from a community-based cohort. PLoS One. 2011;6(11):e27555.
Alric L, Bonnet D, Fort M. Association between female sex, IL28B genotype, but also DQB1*0301 allele and the outcome of acute hepatitis C virus infection. Hepatology. 2014;60(6):2127.
Ikezaki H, Furusyo N, Hiramine S, Ura K, Mitsumoto-Kaseida F, Takayama K, Shimizu M, Toyoda K, Ogawa E, Kainuma M, et al. Association of IL28B rs8099917 genotype and female sex with spontaneous clearance of hepatitis C virus infection: a Japanese cross-sectional study. Arch Virol. 2016;161(3):641–8.
Grebely J, Page K, Sacks-Davis R, van der Loeff MS, Rice TM, Bruneau J, Morris MD, Hajarizadeh B, Amin J, Cox AL, et al. The effects of female sex, viral genotype, and IL28B genotype on spontaneous clearance of acute hepatitis C virus infection. Hepatology. 2014;59(1):109–20.
Moqueet N, Infante-Rivard C, Platt RW, Young J, Cooper C, Hull M, Walmsley S, Klein MB. Favourable IFNL3 genotypes are associated with spontaneous clearance and are differentially distributed in aboriginals in Canadian HIV-hepatitis C co-infected individuals. Int J Mol Sci. 2015;16(3):6496–512.
Peng J, Chen X, He J, Zheng J, Qin B, Jiang Y. Relationship between interleukin 28B, equilibrative nucleoside transporters 1 gene polymorphisms and spontaneous clearance of HCV in HIV/HCV co-infectors. Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi. 2015;36(4):379–82.
Shi X, Pan Y, Wang M, Wang D, Li W, Jiang T, Zhang P, Chi X, Jiang Y, Gao Y, et al. IL28B genetic variation is associated with spontaneous clearance of hepatitis C virus, treatment response, serum IL-28B levels in Chinese population. PLoS One. 2012;7(5):e37054.
Rao HY, Sun DG, Jiang D, Yang RF, Guo F, Wang JH, Liu F, Zhang HY, Zhang HH, Du SC, et al. IL28B genetic variants and gender are associated with spontaneous clearance of hepatitis C virus infection. J Viral Hepat. 2012;19(3):173–81.
Sagnelli E, Santantonio T, Coppola N, Fasano M, Pisaturo M, Sagnelli C. Acute hepatitis C: clinical and laboratory diagnosis, course of the disease, treatment. Infection. 2014;42(4):601–10.
Khakoo SI, Thio CL, Martin MP, Brooks CR, Gao X, Astemborski J, Cheng J, Goedert JJ, Vlahov D, Hilgartner M, et al. HLA and NK cell inhibitory receptor genes in resolving hepatitis C virus infection. Science. 2004;305(5685):872–4.
Dong ZX, Zhou HJ, Xiang XG, Guo SM, Zhuang Y, Zhao GD, Xie Q. IL28B genetic variations are associated with treatment response of patients with chronic hepatitis C in a Chinese Han population. J Dig Dis. 2015;16(2):90–7.
Jin G, Kang H, Chen X, Dai D. Evaluation of the relationship between IL28B, IL10RB and IL28RA single-nucleotide polymorphisms and susceptibility to hepatitis C virus in Chinese Han population. Infec Genet Evol. 2014;21:8–14.
Liao XW, Ling Y, Li XH, Han Y, Zhang SY, Gu LL, Yu DM, Yao BL, Zhang DH, Jin GD, et al. Association of genetic variation in IL28B with hepatitis C treatment-induced viral clearance in the Chinese Han population. Antivir Ther. 2011;16(2):141–7.
Zambrano-Zaragoza JF, Romo-Martinez EJ, Duran-Avelar Mde J, Garcia-Magallanes N, Vibanco-Perez N. Th17 cells in autoimmune and infectious diseases. Int J Inflamm. 2014;2014:651503.
Benoit G, Cooney A, Giguere V, Ingraham H, Lazar M, Muscat G, Perlmann T, Renaud JP, Schwabe J, Sladek F, et al. International Union of Pharmacology LXVI. Orphan nuclear receptors. Pharmacol Rev. 2006;58(4):798–836.
Sun Z, Unutmaz D, Zou YR, Sunshine MJ, Pierani A, Brenner-Morton S, Mebius RE, Littman DR. Requirement for RORgamma in thymocyte survival and lymphoid organ development. Science. 2000;288(5475):2369–73.
Balanescu P, Ladaru A, Voiosu T, Nicolau A, Ene M, Balanescu E. Th17 and IL-17 immunity in chronic hepatitis C infection. Rom J Intern Med. 2012;50(1):13–8.
Chang Q, Wang YK, Zhao Q, Wang CZ, Hu YZ, Wu BY. Th17 cells are increased with severity of liver inflammation in patients with chronic hepatitis C. J Gastroenterol Hepatol. 2012;27(2):273–8.
Sun HQ, Zhang JY, Zhang H, Zou ZS, Wang FS, Jia JH. Increased Th17 cells contribute to disease progression in patients with HBV-associated liver cirrhosis. J Viral Hepat. 2012;19(6):396–403.
Seetharam AB, Borg BB, Subramanian V, Chapman WC, Crippin JS, Mohanakumar T. Temporal association between increased virus-specific Th17 response and spontaneous recovery from recurrent hepatitis C in a liver transplant recipient. Transplantation. 2011;92(12):1364–70.
Kared H, Fabre T, Bedard N, Bruneau J, Shoukry NH. Galectin-9 and IL-21 mediate cross-regulation between Th17 and Treg cells during acute hepatitis C. PLoS Pathog. 2013;9(6):e1003422.
Chen J, Liang Z, Lu F, Fang X, Liu S, Zeng Y, Zhu F, Chen X, Shen T, Li J, et al. Toll-like receptors and cytokines/cytokine receptors polymorphisms associate with non-response to hepatitis B vaccine. Vaccine. 2011;29(4):706–11.
Romero-Gomez M, Eslam M, Ruiz A, Maraver M. Genes and hepatitis C: susceptibility, fibrosis progression and response to treatment. Liver Int. 2011;31(4):443–60.
Selvarajah S, Tobler LH, Simmons G, Busch MP. Host genetic basis for hepatitis C virus clearance: a role for blood collection centers. Curr Opin Hematol. 2010;17(6):550–7.
Ge D, Fellay J, Thompson AJ, Simon JS, Shianna KV, Urban TJ, Heinzen EL, Qiu P, Bertelsen AH, Muir AJ, et al. Genetic variation in IL28B predicts hepatitis C treatment-induced viral clearance. Nature. 2009;461(7262):399–401.
Suppiah V, Moldovan M, Ahlenstiel G, Berg T, Weltman M, Abate ML, Bassendine M, Spengler U, Dore GJ, Powell E, et al. IL28B is associated with response to chronic hepatitis C interferon-alpha and ribavirin therapy. Nat Genet. 2009;41(10):1100–4.
Tanaka Y, Nishida N, Sugiyama M, Kurosaki M, Matsuura K, Sakamoto N, Nakagawa M, Korenaga M, Hino K, Hige S, et al. Genome-wide association of IL28B with response to pegylated interferon-alpha and ribavirin therapy for chronic hepatitis C. Nat Genet. 2009;41(10):1105–9.
Thomas DL, Thio CL, Martin MP, Qi Y, Ge D, O'Huigin C, Kidd J, Kidd K, Khakoo SI, Alexander G, et al. Genetic variation in IL28B and spontaneous clearance of hepatitis C virus. Nature. 2009;461(7265):798–801.
Gaudieri S, Rauch A, Park LP, Freitas E, Herrmann S, Jeffrey G, Cheng W, Pfafferott K, Naidoo K, Chapman R, et al. Evidence of viral adaptation to HLA class I-restricted immune pressure in chronic hepatitis C virus infection. J Virol. 2006;80(22):11094–104.
Ansari MA, Pedergnana V, LCI C, Magri A, Von Delft A, Bonsall D, Chaturvedi N, Bartha I, Smith D, Nicholson G, et al. Genome-to-genome analysis highlights the effect of the human innate and adaptive immune systems on the hepatitis C virus. Nat Genet. 2017;49(5):666–73.
Moore CB, John M, James IR, Christiansen FT, Witt CS, Mallal SA. Evidence of HIV-1 adaptation to HLA-restricted immune responses at a population level. Science. 2002;296(5572):1439–43.
von Delft A, Humphreys IS, Brown A, Pfafferott K, Lucas M, Klenerman P, Lauer GM, Cox AL, Gaudieri S, Barnes E. The broad assessment of HCV genotypes 1 and 3 antigenic targets reveals limited cross-reactivity with implications for vaccine design. Gut. 2016;65(1):112–23.
Thimme R, Bukh J, Spangenberg HC, Wieland S, Pemberton J, Steiger C, Govindarajan S, Purcell RH, Chisari FV. Viral and immunological determinants of hepatitis C virus clearance, persistence.and disease. Proc Nati Acad Sci USA. 2002;99(24):15661–8.
Thimme R, Oldach D, Chang KM, Steiger C, Ray SC, Chisari FV. Determinants of viral clearance and persistence during acute hepatitis C virus infection. J Exp Med. 2001;194(10):1395–406.
Klenerman P, Thimme R. T cell responses in hepatitis C: the good the bad and the unconventional. Gut. 2012;61(8):1226–34.
Basha HI, Subramanian V, Seetharam A, Nath DS, Ramachandran S, Anderson CD, Shenoy S, Chapman WC, Crippin JS, Mohanakumar T. Characterization of HCV-specific CD4+Th17 immunity in recurrent hepatitis C-induced liver allograft fibrosis. Am J Transplant Off J Am Soc Transplant Am Soc Transplant Surg. 2011;11(4):775–85.
Wang JM, Shi L, Ma CJ, Ji XJ, Ying RS, Wu XY, Wang KS, Li G, Moorman JP, Yao ZQ. Differential regulation of interleukin-12 (IL-12)/IL-23 by Tim-3 drives T(H)17 cell development during hepatitis C virus infection. J Virol. 2013;87(8):4372–83.
Shen T, Chen X, Zhang W, Xi Y, Cao G, Zhi Y, Wang S, Xu C, Wei L, Lu F, et al. A higher correlation of HCV core antigen with CD4+ T cell counts compared with HCV RNA in HCV/HIV-1 coinfected patients. PLoS One. 2011;6(8):e23550.
Jauncey M, Micallef JM, Gilmour S, Amin J, White PA, Rawlinson W, Kaldor JM, van Beek I, Dore GJ. Clearance of hepatitis C virus after newly acquired infection in injection drug users. J Infect Dis. 2004;190(7):1270–4.
Paradowska-Gorycka A, Stypinska B, Pawlik A, Romanowska-Prochnicka K, Haladyj E, Manczak M, Olesinska M. RORC2 genetic variants and serum levels in patients with rheumatoid arthritis. Int J Mol Sci. 2016;17(4):488.
Acknowledgements
We are grateful to all who provided blood samples for this study. We also thank Dr. Weidong Zhang and Dr. Yuanlin Xi for their assistance for construction of the study cohort.
Funding
This work was financially supported by the National Science and Technology Major Project of China (2017ZX10202101-003, 2018ZX10731101-001-008 and 2018ZX09739002-003), State Key Laboratory of Infectious Disease Prevention and Control (2015SKLID506) and the National Natural Science Foundation of China (81271826, 81201286).The funding bodies had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.
Availability of data and materials
All data generated and analyzed during this study are included in this published article and its Additional files. The datasets analyzed during the current study are available from the corresponding author on request.
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Contributions
ZX, YL and LL evaluated the data, performed analyses, and contributed to writing the manuscript. YL contributed the cellular total DNA isolation and purification. WC and HL participated in the study design. TS conceived of the study, also involved in the evaluation and interpretation of data, and wrote the manuscript. All authors read and approved the final manuscript.
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Ethics approval and consent to participate
The study was approved by the institutional review authorities of Peking University Health Science Center (Approval ID: PKUPHLL20090011). All patients provided written informed consent before enrollment in the study.
Competing interests
The authors declare that they have no competing interests.
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Additional file
Additional file 1:
Figure S1. Comparison of distributions of IFNL3 and RORC SNP genotypes among HIVneg, HIVpos, and total resolved individuals. (a) Distribution of genotypes (%) of three IFNL3 SNPs (rs12979860, rs8099917, and rs12980275). (b) Distribution of genotypes (%) of two RORC SNPs (rs9826, and rs1521177). Chi-square (χ2) and Fisher’s exact tests were used to evaluate the differences in SNP distributions between two groups. P-values (two-tailed) < 0.05 were considered significant (n.s., not significant). Figure S2. HCV viral load levels of RORC SNP genotypes (rs9826, rs1521177) among HIVneg HCV carriers of the primary cohort. Figure S3. Linkage disequilibrium tests for RORC SNPs (rs9826 and rs1521177) and IFNL3 SNPs (rs12979860, rs8099917 and rs12980275) in the primary cohort (a) D’ value. (b) r2 value. Table S1. Allele frequencies of IFNL3 gene SNPs in different populations. Table S2. Allele, genotype, and carrier frequencies and percentages of tested SNPs in the primary cohort. Table S3. RORC gene SNP allele frequencies in different populations. Table S4. The Hardy–Weinberg Equilibrium (HWE) test of all SNPs in the whole study population. Table S5. Genotype and allele frequency distributions of RORC SNPs in HIVpos group. Table S6. Genotype and allele frequency distributions of RORC SNPs in HIVneg HCV carriers of the primary cohort. Table S7. Linkage disequilibrium tests for RORC rs9826/rs1521177 in the primary cohort and the IFNL3 favorable sub-cohort. (DOC 1671 kb)
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Xie, Z., Li, Y., Long, L. et al. Polymorphisms in the Th17 cell-related RORC gene are associated with spontaneous clearance of HCV in Chinese women. BMC Infect Dis 18, 254 (2018). https://doi.org/10.1186/s12879-018-3153-2
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DOI: https://doi.org/10.1186/s12879-018-3153-2