Genetic Susceptibility Study Of Immune Response To Hepatitis B Vaccination

BackgroundsInfection with hepatitis B virus is (HBV) is a public health problem that seriously threatens human life. Approximately93million Chinese people are carriers of HBV, and300,000deaths occur every year in China as a result of HBV infection. Recombinant vaccines against HBV have been used since the1980s and are an effective method of preventing HBV infection and transmission. However,5-10%of healthy adults fail to produce protective levels of antibody against the hepatitis B vaccine (anti-HBs). The differed outcomes of immune response to hepatitis B vaccination bring out difficulties in HBV prevention. Up to now, several studies have documented that genetic polymorphisms were associated with immune response to hepatitis B vaccination. In the present study, case-control study was used to evaluate the potential SNPs association with immune response to hepatitis B vaccination.MethodsIndividuals from Beijing and Shandong Province were administered three doses of hepatitis B vaccine at0,1, and6months. In Beijing population, a total of10single nucleotide polymorphisms distributed in6cytokine and cytokine receptor genes (TNFRSF1A, IL12A, IL12B, IFNG, IL4, and IL10) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), polymerase chain reaction with sequence-specific primers (PCR-SSP), and TaqMan-MGB probe-based real-time polymerase chain reaction, among214high-responders (anti-HBs>1,000mIU/ml) and107low-responders (anti-HBs:10-99mIU/ml) to three dose of hepatitis B vaccine.A total of111single nucleotide polymorphisms (SNPs) in17genes that encodes T cell co-receptors and costimulatory molecules, were analyzed using the iPLEX MassARRAY system, among214high-responders and107low-responders to three dose of hepatitis B vaccine. The SNPs with P values<0.05were selected for follow-up analysis with the TaqMan-MGB or TaqMan-BHQ probe-based real-time polymerase chain reaction in Shandong population, which included1090high-responders and636-low responders to three dose of hepatitis B vaccine.A Genome Wide Association Study (GWAS) of non-response to hepatitis B vaccination was performed using Illumina_HumanOmniExpress SNP Array among78non-responders to six dose of hepatitis B vaccine and113high responders to three dose of hepatitis B vaccine. The SNPs with P-correction<0.05(FDR_BH multiple correction) were selected for follow-up analysis with the TaqMan-MGB or TaqMan-BHQ probe-based real-time polymerase chain reaction among high responders and non-responders to three dose of hepatitis B vaccine in Shandong (1122vs.374) and Beijing (214vs.46) population.Results1. Association of polymorphisms in cytokine and cytokine receptor genes with immune response to hepatitis B vaccinationThe minor allele'CTCTAA; of rs17860508in the IL12B gene was associated with a low response to hepatitis B vaccination (P=0.039, odds ratio=1.41,95%confidence interval=1.00-1.99). In addition, a significant gene-gene interaction was found:the frequency of the combined genotypes (IL12A rs2243115'TT'and IL12B rs17860508'CTCTAA/CTCTAA') was significantly higher in the low-response group than in the high-response group(P=0.008, odds ratio=2.19,95%confidence interval=1.23-3.93).2. Association of polymorphisms in genes that encode T cell co-receptors and costimulatory molecules with immune response to hepatitis B vaccinationThree SNPs (rs12133337and rs10918706in the CD3Z gene, rs10912564in the OX40L gene) were associated significantly with the immune response to hepatitis B vaccination in Beijing population (P=0.008,0.041, and0.019, respectively). Confirmation study of these three SNPs was performed in Shandong population, the minor allele'C'of rs12133337continued to show an association with a lower response to hepatitis B vaccination (P=0.033, odds radio=1.28,95%confidence interval=1.01-1.61). Furthermore, in the stratified analysis for both the Beijing and Shandong populations, the association of the minor allele'C' of rs12133337with a lower response to hepatitis B vaccination was more prominent after individuals who were overweight or obese (body mass index≤25kg/m2) were excluded (Beijing population:P=0.003; Shandong population:P=0.002).3. GWAS of non-response to hepatitis B vaccinationTwenty-seven SNPs in HLA-DR region were associated with non-response to booster hepatitis B vaccination after FDR_BH multiple corrections (P<0.05). Linkage disequilibrium analysis showed that other23SNPs were tagged by4SNPs (rs477515, rs3763316, rs28366298, rs13204672). Confirmation studies of these four SNPs were performed in Shandong and Beijing primary immune populations. These four SNPs were also associated with non-response to hepatitis B vaccination in both the Shandong and Beijing populations (Shandong population:P=2.82e-010-9.44e-07; Beijing population: P=1.58e-04-0.048).Conclusions1. The minor allele'CTCTAA'of rsl7860508in the IL12B gene is associated with susceptibility to a low response to hepatitis B vaccination. In addition, interaction between rs2243115in IL12A and rs17860508in IL12B enhances the risk of a low response to the hepatitis B vaccination.2. Polymorphism (rsl2133337) in the CD3Z gene might affect the immune response to hepatitis B vaccination, and that a lower BMI might increase the contribution of the polymorphism to immunity to hepatitis B vaccination.3. Four SNPs (rs477515, rs28366298, rs3763316, rsl3204672) located in HLA-DR region were significantly associated with non-response to hepatitis B vaccination; Haplotype'CACA'was a protective factor in immune response to hepatitis B vaccination; Polymorphisms in the HLA-DR region might affect the immune response to hepatitis B vaccination.