Correlation Of Plasma Estradiol And Interleukin 12 Levels In HCV Infected Patients And Association Of Estrogen Receptor 2 Polymorphisms With Hepatitis C Virus Infection Outcomes In A High-risk Chinese Han Population

Section 1 Correlation of plasma estradiol and interleukin 12 levels in HCV infected patients[Objective] In the late 1980 s to 90 s, some nonstandard and illegal commercial blood centers were existed in regions of Jiangsu province, caused spread of blood transmission virus such as hepatitis C virus(HCV). The Hepatitis C caused by HCV infection is easily develops chronic infection, ranging from minimal inflammation to cirrhosis and hepatocellular carcinoma, which seriously influcend their life quality and survive rate. To determine the correlation of plasma estradiol(E2) and cytokines levels such as interleukin 12(IL-12) in HCV infected paid donors.[Methods] The study was conducted among 199 paid donors. All participants were divided into three groups: 72 healthy controls, 39 spontaneous clearances and 88 persistent infection subjects. We performed all participates’ plasma E2 level, IL-12 concentration and liver function.[Results] The history of paid blood donation, the frequency of blood donation, alanine aminotransferase and aspartate aminotransferase were significantly different among the three groups(all P < 0.05). Multiple logistic regression analysis showed that the history of full blood donation(OR = 2.83, 95% CI = 1.05-7.70) and the history of plasma donation(OR = 5.57, 95% CI = 2.26-13.75) were significantly associated with HCV infection. Furthermore, the plasma E2 concentration was positively correlated with IL-12 level(r = 0.654,P < 0.001) among HCV infected person.[Conclusion] Paid blood donation is a risk factor of HCV infection. Our study indicates estradiol may positively regulate the expression of IL-12, and then affect HCV infection outcomes.Section 2 Association of estrogen receptor 2 polymorphisms with hepatitis C virus infection outcomes in a high-risk Chinese Han population[Background] Hepatitis C virus(HCV) has different clinical and biological characteristics in women versus men, which suggests the potential involvement of estrogen. Estrogen signaling is mediated by the estrogen receptor(ER), and genetic variations in the estrogen receptor gene(ESR) might affect the pathology of HCV infection. However, it is currently unclear whether potentially functional single nucleotide polymorphisms(SNPs) within ESR affect the development and outcomes of HCV infection.[Methods] This population-based, case-control study was conducted with 813 patients with persistent infection, 479 patients with spontaneous clearance and 1051 controls. After informed consents from all the participants, question aires were used to collect epidemiological data, and blood samples were collected for genotyping. We performed logistic regression analysis to explore the associations between three potentially functional SNPs(rs1256049, rs4986938 and rs944459) of estrogen receptor 2(ESR2) and HCV infection outcomes by calculating the odds ratio(OR) and its 95% confidence interval(CI). We selected the Dual-Luciferase reporter assay system as carrier, and build the reporter gene plasmids of ESR2 containing different SNPs. Then these plasmids were transiently transfected into HEK293 and He La cell. The relative luciferase activity was calculated to explore the role of different SNPs on transcriptional activities of different 3ˊ-untranslated region(3ˊ-UTRs).[Results] Male carriers of the SNP rs4986938 A allele were significantly associated with increased susceptibility to HCV infection after adjustment for age and route of infection(additive model: adjusted OR = 1.493, 95% CI = 1.102-2.022, P = 0.010). The A allele of rs4986938 had a significantly reduced risk of HCV infection in the female subgroup(GA vs. GG: adjusted OR = 0.710, 95% CI = 0.544-0.928, P = 0.012; dominant model: adjusted OR = 0.686, 95% CI = 0.530-0.889, P = 0.004; additive model: adjusted OR = 0.703, 95% CI = 0.560-0.883, P = 0.002). In addition, females carrying the AA genotype appeared to clear HCV spontaneously more readily(adjusted OR = 0.237, 95% CI = 0.078-0.716, P = 0.011), and additive model analyses showed that each additional allele contributed a decreased risk of approximately 34% for HCV chronicity(adjusted OR = 0.659, 95% CI = 0.489-0.889, P = 0.006). Furthermore, a significant multiplicative interaction between the combined rs1256049 and rs4986938 genotypes was found to decrease HCV infection risk(adjusted OR = 0.583, 95% CI = 0.436-0.780, P = 3.000×10-4). The area under the curve(AUC), based on the model and including the factors of age, gender and HCV genotypes, was not significantly related to the clearance of HCV(95% CI = 0.496-0.560,P = 0.092). Considering the three genotypes, the model for viral clearance prediction with the above factors produced an AUC of 0.550(95% CI = 0.518-0.582, P = 0.003). As compared with the wild plasmid, the mutational plasmids had significantly increased 3ˊ-UTR activity in both cells.[Conclusions] We provide here the first report that rs4986938 in the ESR2 gene played a potential sex-specific role in the etiology of HCV infection in a high-risk Chinese Han population, suggesting that ESR2 is a candidate susceptibility gene for HCV infection and viral clearance.