Effects Of Maternal Anti-HBs On Infants Immunne Response To Hepatitis B Vaccine And The Association Between Polymorphisms Of Hepatitis C Virus NS3 Proteins And Liver Cirrhosis

Part I Seroprevalence of HBsAg among pregnant women in multi-centers in ChinaObjective To investigate the prevalence of hepatitis B surface antigen (HBsAg) among pregnant women in ChinaMethods The investigation was divided into three parts:Part I was conducted from January 2010 to December 2015, pregnant women were screened HBsAg in Hubei province(Dangyang city, Daye city, Huanggang city, Xiaogan city, Yicheng city, Badong country, Tongcheng country), Xinjiang uygur autonomous region (Yining city, Shihezi city), Shanxi province(Taiyuan city, Yuncheng city), Guangdong province(Taishan city), Qinghai province and Hainan province to survey epidemics of HBsAg; Part Ⅱ was conducted from March 2012 to December 2015, pregnant women were tested HBV markers in Hubei province, and detect HBV-DNA in HBsAg positive pregnant women. In Part Ⅲ, HBsAg positive pregnant women form Shanxi province and Hubei province in part I and part Ⅱ were enrolled and tested HBV markers as well as HBV-DNA, in orlder to analyze the status of HBV replication.Results Among 324138 pregnant women in part Ⅰ, HBsAg positive rate was 5.8%. Pregnant women in Qinghai province own the lowest HBsAg positive rate (2.6%), and pregnant women in Hainan province own the highest HBsAg positive rate (13.4%), the prevalence of HBsAg of pregnant women in Xinjiang uygur autonomous region, Shanxi province, Hubei province and Guangdong province was 4.7%,4.2%, 6.6% and 8.5%, respectively. Among 1584 pregnant women of Hubei province in part Ⅱ, the prevalence of HBsAg was 7.1%; Among 1190 HBsAg positive pregnant women in part Ⅲ,38.7% of them were positive for HBeAg, the detection rate and titers of HBV DNA was higher in HBeAg positive women when compared to HBeAg negative women.Conclusion The multicenter study shows that positive rate of HBsAg was high among pregnant women and the prevalence HBsAg was different among regions in China. It is necessary to continue to strengthen the screening of maternal infection HBV, in order to further improve mother-to-child transmission.Part Ⅱ Effects of maternal anti-HBs on the immune response of infants to hepatitis B vaccineObjective Hepatitis B vaccination is the most effective way to prevent HBV infection. Although the protective effect of HB vaccine was remarkable, anti-HBs titers of 5% to 15% children failed to reach the levels necessary for protection (anti-HBs titers ≥10IU/L) even after completing 0-,1- and 6-month scheme. It is still controversial whether maternal anti-HB antibodies (anti-HBs) affect infants’ immune response to HB vaccination. A multi-center study conducted aims to address this question.Methods A multi-center prospective study was conducted from March 2012 to November 2015 in Zhongnan hospital of Wuhan University, Mother and Child Hospital of Wuxue city, Centers for Disease Control and Prevention of Chongyang country, Centers for Disease Control and Prevention of Xiaonan District, Centers for Disease Control and Prevention of Guangshui city, Centers for Disease Control and Prevention of Chibi city. The enrolled subjects were HBsAg-negative pregnant women and their infants of 7-24 months old who received HB vaccines. In total,1063 mother-and-infant pairs were enrolled. For determining the transplacental transfer of maternal anti-HBs, pregnant women were recruited between March to April in 2012 in the department of Obstetrics and Gynecology of Zhongnan hospital of Wuhan University. Total 90 mother-and-infant pairs were enrolled.Results The subjects were divided into three groups by maternal anti-HBs titers of 10-99 IU/L (26,29%),100-499 IU/L (29,32%) and≥500 IU/L (35,39%). The transplacental transfer rate of anti-HBs in the three groups was 100%,100% and 89.7%, respectively (P<0.05). The transplacental transfer rate of anti-HBs was higher in the group 100-499 IU/L and≥500 IU/L when compared to the group 10-99 IU/L. Total 1063 mother-and-infant pairs were enrolled and divided into three groups by maternal anti-HBs titers of<10 IU/L (negative),10-499 IU/L and≥500 IU/L, with 62.1% of these mothers were positive. The infants’ anti-HBs positive rate and titers were negatively correlated with the maternal anti-HBs titers:the anti-HBs positive rate of infants were 88.9%(360/405),84.5%(381/451) and 77.3%(160/207), respectively,p<0.0001. Median titers of anti-HBs (IU/L) among infants were 169.1, 141.0 and 79.4 respectively,p=0.020. One hundred and sixty-two infants were negative for anti-HBs after the standard vaccination, and 120 out of 131 of these infants (91.6%) reached anti-HBs-positivity after the first catch-up dose. The maternal anti-HBs titers did not significantly affect infant response to this catch-up dose.Conclusion High titers of maternal anti-HBs may suppress immune response of infants to the standard HB vaccination. The current schedule of the 0-,1- and 6-month may not be the optimal choice of infants with anti-HBs positive mothers.Part Ⅲ Exploring the effects of anti-HBs on the antigenicity of recombinant HBsAg in vitroObjective Anti-HBs could neutralize HBsAg, but whether affect its antigenicity is unclear.Methods A study was conducted in Zhongnnan hospital from March 2016 to April 2016, PBMC were extracted from 8 healthy persons who was positive for anti-HBs and treated with rHBsAg and rHBsAg plus anti-HBs, lymphocytes proliferation rate, levels of IL-10, IL-4 and IFN-γ were measured.Results Lymphocytes proliferation rate was higher in PBMC treated with rHBsAg when compared with rHBsAg plus anti-HBs, the lymphocytes proliferation rate increased as the anti-HBs concentration decreased when cells treated with rHBsAg plus anti-HBs. Compared with treated with rHBsAg, the lymphocytes proliferation rate was significantly lower when the anti-HBs concentration were 10000IU/L and 1000 IU/L (P=0.002, P=0.045). The lymphocytes proliferation was not significant when treated with anti-HBs.The levels of IL-10 was higher in PBMC treated with rHBsAg when compared with rHBsAg plus anti-HBs, the levels of IL-10 increased as the anti-HBs concentration decreased when cells treated with rHBsAg plus anti-HBs. Compared with treated with rHBsAg, the levels of IL-10 was significantly lower when the anti- HBs concentration were 10000IU/L and 1000 IU/L (P<0.0001, P<0.0001). The levels of IL-10 was not significant when treated with anti-HBs.The levels of IL-4 was higher in PBMC treated with rHBsAg when compared with rHBsAg plus anti-HBs, the levels of IL-4 increased as the anti-HBs concentration decreased when cells treated with rHBsAg plus anti-HBs. Compared with treated with rHBsAg, the lymphocytes proliferation rate was significantly lower when the anti-HBs concentration was 10000 IU/L (P=0.012). The lymphocytes proliferation was significant when treated with anti-HBs.The levels of IFN-γ was higher in PBMC treated with rHBsAg when compared with rHBsAg plus anti-HBs, the levels of levels of IFN-γ increased as the anti-HBs concentration decreased when cells treated with rHBsAg plus anti-HBs. Compared with treated with rHBsAg, the levels of IFN-y was significantly lower when the anti-HBs concentration were 10000IU/L and 1000 IU/L (P<0.0001, P=0.0005). The levels of IFN-y was not significant when treated with anti-HBs.Conclusion The antigenicity of recombinant HBsAg could be inhibited by anti-HBs in vitro, and could inhibit the activation of Thl cells (secrete IL-10) and Th2 cells (secrete IL-4 and IFN-y), and the inhibition was enhanced by increasing of anti-HBs’s concentration.Part IV Investigation the prevalence of hepatitis c virus among paid blood donors in part area of hubei provinceObjective Paid blood donation is an important way of HCV infection. A study was conducted in order to investigate the prevalence of hepatitis c virus and the distribution of HCV genotype.Methods Form March 2014 to June 2015, paid blood donors were enrolled in Centers for Disease Control and Prevention of Wuxue city, Health Centre of Dougang Town in Xiaogan city and Health Centre of Yandian Town in Xiaogan city to test anti-HCV, only anti-HCV positive patients were enrolled to detect HCV-RNA. HCV-RNA titer was measured and HCV genotype was determined by sequencing among these patients who have detectable HCV-RNA.Results A total of 252 paid blood donors were enrolled, the average age of them were 54±6.9 years, including 46 men and 206 women. One hundred and fifty-seven patients(62.3%,157/252) were positive for anti-HCV, and 110 (70.1%,110/157) of them owned detectable HCV-RNA, the average titers of them were 5.8 ± 1.0 (log). Ninety-night (89.1%,98/110) of them were successful genotyped, among these patients,82 of them were HCV genotype 1b and 16 of them were HCV genotype 2a.Conclusion The prevalence of hepatitis c virus among paid blood donors in some part area of Hubei provience was high, the dominant genotype was HCV genotype 1 b. Only HCV genotype 1b was founded in Wuxue city, but both HCV genotype 1b and 2a were founded in Xiaogan city. There are no other genotypes was founded in this study.Part V Association between Polymorphisms of the NS3 Proteins of Hepatitis C Virus Genotype lb and Liver CirrhosisObjective Polymorphisms of the hepatitis C virus (HCV) core protein and NS3 have been described to be associated with liver cirrhosis (LC) and/or hepatocellular carcinoma in patients with chronic hepatitis C genotype lb (HCV lb). Here, we determine whether there is an association between LC and polymorphisms of viral core protein and NS3 in patients in Hubei province, China.Methods A total of 42 patients with chronic HCV lb (21 patients with LC and 21 with chronic hepatitis [CH]) were enrolled, amino acid sequence of the core and NS3 were obtained by direct sequencing and compared between patients with LC and patients CH.Results No significant difference of amino acid polymorphisms was observed between isolates from LC and CH patients in the core protein. However, amino acid polymorphisms Thr 1072, Val1074 and None-Thr1098 were correlated significantly with LC.Conclusion Polymorphisms of Thr 1072, Val1074 and None-Thr1098 in NS3 may be the risk factors for the development of LC in patients with chronic HCV 1 b.