| Objectives:1.To investigate the prevalence of intrauterine transmission of hepatitis B virus(HBV)among hepatitis B surface antigen(HBsAg)positive pregnant women in Wuhan,Hubei province;to investigate the influencing factors of HBV intrauterine transmission,and to identify the key risk factors.2.To investigate the associations between maternal genetic variants and risk of HBV intrauterine transmission,and to identify the susceptibility genes and loci of HBV intrauterine transmission.3.To investigate the associations between neonatal genetic variants and risk of HBV intrauterine infection,and to identify the susceptibility genes and loci of HBV intrauterine infection.4.To explore the gene-environment interactions in modifying susceptibility to HBV intrauterine infection.Methods:1.In the first part,HBsAg positive pregnant women and their neonates were consecutively enrolled in this study from May 2012 to May 2016 at Wuhan Women and Children Medical and Healthcare Center,Wuhan,China.Epidemiological survey was carried out to investigate the influencing factors of HBV intrauterine infection.Independent-samples t test,Pearson chi-square test,trend chi-square test,Fisher's exact test,univariate and multivariate unconditional Logistic regression analysis were conducted to evaluate the risk factors of HBV intrauterine infection.2.In the second and third part,a case-control study was conducted.81 pregnant women with HBV intrauterine transmission and their neonates were classified as case group,and 162 pregnant women delivering during the same period who were free from HBV intrauterine transmission and their neonates were classified as control group.Maternal candidate single nucleotide polymorphisms(SNPs)in key genes of LT?R/APOBEC3B,PD-1/PD-L1 and TRAIL/TRAIL-R2 pathway and neonatal candidate SNPs in genes encoding important pattern recognition receptors(TLR9,RIG-1,MDA5 and cGAS)and key adaptor proteins(MyD88,MAVS and STING)in the innate immune response were genotyped using MiSeq genotyping platform.Independent-samples t test,Pearson chi-square test,trend chi-square test,Fisher's exact test,Spearman rank correlation test,univariate and multivariate unconditional Logistic regression analysis,decision tree analysis,and cumulative effect analysis were applied to evaluate the associations of each SNP of mothers and neonates with risk of HBV intrauterine infection.3.In the fourth part,mothers from the second part and neonates from the third part were included in the analysis.The interactions between gene(maternal and neonatal susceptibility loci)and environment(maternal HBeAg during pregnancy and mode of delivery)were calculated using multivariate unconditional Logistic regression analysis and the excel sheet made by Andersson et al.in both additive and multiplicative model.Results:1.During the study period,969 neonates born to HBsAg positive mothers had serum HBsAg and HBV DNA detected,among whom,81 were infected with HBV intrauterinely.The HBV intrauterine infection rate was 8.36%(81/969).The results of multivariate unconditional Logistic regression analysis showed that:maternal HBeAg positivity during pregnancy(OR=2.31,95%CI:1.02-5.20)was an independent risk factor of HBV intrauterine transmission,while cesarean section(OR =0.34,95%Cl:0.19-0.58)was significantly associated with decreased risk of HBV intrauterine infection.Mothers with medium(6.00-7.99 log,o copies/mL)and high(?8.00 log10 copies/mL)HBV DNA level in the third trimester had a 3.21-fold(OR = 3.21,95%CI:1.38-7.43)and a 6.05-fold(OR=6.05,95%CI:1.91-19.15)increased risk of HBV intrauterine transmission compared to those with low(<6.00 log10 copies/mL)HBV DNA level,and statistical significance was also observed in the trend test(P= 0.001).2.The associations of each maternal SNP with HBV intrauterine transmission were evaluated in the second part,and the results showed that:after adjusting for maternal HBeAg and mode of delivery,mothers who carried the rs2239704 TT genotypes in LTA gene had a higher risk of HBV intrauterine transmission compared to those who carried GG genotypes(OR=2.94,95%CI:1.03-8.41);a significantly increased risk of HBV intrauterine transmission was also found under the additive model(OR=1.66,95%CI:1.04-2.64).Under the additive model,maternal PDCD1 rs2227981 and TNFRSF10B rs1129424 T allele carriage were associated with a 45%(OR=0.55,95%CI:0.32-0.95)and a 55%(OR=0.45,95%CI:0.21-0.98)decreased risk of HBV intrauterine transmission,respectively.Moreover,the results of gene-gene interaction analysis indicated that a high-order interaction existed among maternal PDCD1 rs2227981,LTBR rs3759333 and CD274 rs822336.Furthermore,the accumulative effects of maternal LTA rs2239704,PDCD1 rs2227981 and TNFRSF10B rs1129424 were observed(P for trend=0.008);mothers who carried 2-3 risk genotypes had a significantly increased risk of HBV intrauterine transmission when compared to those who did not carry any risk genotypes(OR=4.38,95%CI:1.68-11.44).3.The associations between each neonatal SNP and HBV intrauterine infection were e-valuated in the third part,and the results revealed that:after adjusting for maternal HBeAg and mode of delivery,neonates who carried the rs3746661 CG and CG/CC genotypes in MAVS gene had a 159%(OR=2.59,95%Cl:1.37-4.90)and a 151%(OR=2.51,95%CI:1.36-4.65)increased risk of HBV intrauterine infection compared to those who carried GG genotypes.Under the additive model,neonatal MAVS rs3746661 C allele and rs8126207 G allele carriage were associated with a 87%(OR=1.87,95%CI:1.16-3.02)and a 84%(OR=1.84,95%CI=1.06-3.20)increased risk of HBV intrauterine infection,respectively.Moreover,the results of gene-gene interaction analysis showed that a significant interaction existed between neonatal MAVS rs3746661 and MB21D1 rs311678.Furthermore,the accumulative effects of neonatal MAVS rs3746661 and rs8126207 were observed(P for trend=0.002);neonates who carried one risk genotype and two risk genotypes had a 180%(OR=2.80,95%Cl:1.20-6.52)and a 323%(OR=4.23,95%Cl:1.71-10.43)increased risk of HBV intrauterine infection when compared to those who did not carry any risk genotypes.4.Gene-environment interactions analysis showed that interactions of maternal LTA rs2239704(AP=0.75,95%Cl:0.17-1.33)and TNFRSF10B rs1129424(AP=0.76,95%Cl:0.21-1.30)collaborating with maternal HBeAg during pregnancy to modify the risk of HBV intrauterine transmission.Moreover,a significant multiplicative interaction(Pmul=0.044)and additive interaction(RERI=2.03,95%CI:0.29-3.76;AP=0.74,95%CI:0.33-1.14)were observed between neonatal MAVS rs8126207 and mode of delivery in modifying the risk of HBV intrauterine infection.Conclusions:1.The prevalence of HBV intrauterine transmission among HBsAg positive pregnant women was 8.36%in Wuhan,Hubei province.Maternal HBeAg positivity during pregnancy and high HBV DNA loads in the third trimester were two key risk factors of HBV intrauterine infection,while cesarean section might be a protective factor of HBV intrauterine infection.2.Maternal LTA rs2239704,PDCD1 rs2227981and TNFRSF10B rs1129424 were significantly associated with the risk of HBV intrauterine transmission.3.Neonatal MAVS rs3 746661 and rs8126207 were significantly associated with the risk of HBV intrauterine infection.4.The two maternal genetic variants LTA rs2239704,TNFRSF10B rs1129424 interacting with maternal HBeAg during pregnancy to influence the risk of HBV intrauterine transmission.Neonatal MAVS rs8126207 collaborating with mode of delivery to modify the risk of HBV intrauterine infection. |
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