乙型肝炎疫苗初次免疫成年正常应答和高应答者3年抗体持久性观察

Anti-HBs persistence following primary vaccination with three doses of hepatitis B vaccine among normal and high-responder adults：a 3-year follow-up study

目的：探讨乙型肝炎疫苗（HepB）初次免疫成年正常应答和高应答者3年的抗体持久性及其影响因素。方法于2009年9月，选择山东省章丘市北部相邻的3个人口流动性较小的乡镇，在其全部79个村中选择既往无乙型肝炎病毒（HBV）感染史和/或HepB免疫史、居住6个月以上、健康状况良好的18~49岁人群，共24237名。采集静脉血3~5 ml，采用ELISA法检测乙型肝炎表面抗原（HBsAg）、乙型肝炎表面抗体（抗-HBs）和乙型肝炎核心抗体（抗-HBc）。经检测，以上3项指标均为阴性者11590名，按照以村为单位整群随机抽样方法，将其分为4组，按照0-1-6免疫程序分别接种3剂次20μg重组酵母HepB（HepB-SC）、20μg重组中国仓鼠卵巢细胞（HepB-CHO）、10μg HepB-SC和10μg重组汉逊酵母HepB（HepB-HP）进行初次免疫，对其中的6386名正常应答（2869名）和高应答（3517名）者进行随访（调查基本情况、HBV感染史、HepB免疫史、吸烟史、饮酒史、慢性病史等），共随访4677名，随访率为73.24%。于初次免疫后1个月（T1）和3年（初免后3年）采集血标本，采用化学发光微粒子免疫分析法定量检测抗-HBs、HBsAg和抗-HBc。分别采用多因素非条件logistic回归模型和多因素线性回归模型分析抗-HBs阳性和抗-HBs几何平均浓度（GMC）的影响因素。结果4677名随访对象中，男性占43.06%（2014名），女性占56.94%（2663名）。T1时抗-HBs阳性率为100%，初免后3年为80.99%（3788例）；T1时抗-HBs的GMC为1413.48（95%CI：1358.86～1470.30）mU/ml，初免后3年为60.33（95%CI：56.97～63.90）mU/ml。与初次免疫接种20μg HepB-CHO者相比，接种20μg HepB-SC、10μg HepB-SC和10μg HepB-HP者初免后3年抗-HBs阳性率较低，OR（95%CI）值分别为0.65（0.50～0.84）、0.52（0.41～0.67）和0.31（0.28～0.45），其抗-HBs的GMC亦较低，b（95%CI）值分别为-0.33（-0.47～-0.20）、-0.41（-0.55～-0.28）和-0.78（-0.92～-0.65）；与初次免疫接种年龄为18~29岁者相比，30~39和40~49岁者初免后3年抗-HBs的GMC较低，b（95%CI）值分别为-0.31（-0.47～-0.15）和-0.24（-0.39～-0.09）；与T1时抗-HBs的GMC为100~999 mU/ml者相比，GMC为1000~1999、2000～2999和≥10000 mU/ml者抗-HBs阳性率较高，OR（95%CI）值分别为4.97（3.80～6.49）、7.87（16.19～10.01）和9.67（6.47～14.44），其抗-HBs的GMC亦较高，b（95%CI）值分别为1.00（0.87～1.14）、1.85（1.74～1.97）和3.28（3.12～3.44）。初免后3年HBsAg阳性者4例，抗-HBc阳转率为4.68%（219/4677）。结论 HepB初次免疫成年正常应答和高应答者初免3年后，抗-HBs的GMC下降，但抗-HBs的阳性率仍维持在较高水平，保护水平较强；初免3年后抗体持久性主要与初次免疫疫苗类型、年龄和T1时抗-HBs的GMC有关。

Objective To assess the 3-year anti-HBs persistence after primary vaccination with three-dose of hepatitis B vaccine （HepB） among normal and high-responder adults. Methods A total of 24 237 healthy adults who had no histories of hepatitis B infection and hepatitis B vaccination, resided in local areas for more than six months and were aged 18-49 years were selected from 79 villages of Zhangqiu county, Shandong province, China in 2009. Blood samples were obtained and hepatitis B surface antigen （HBsAg）, antibody against hepatitis B surface antigen （anti-HBs） and antibody against hepatitis B core antigen （anti-HBc） were detected using ELISA method. A total of 11 590 persons who were negative for all of these indicators were divided into four groups by cluster sampling method. Each group was vaccinated with one of the following four types of HepB at 0-1-6 months schedule： 20 μg HepB derived in Saccharomyces cerevisiae （HepB-SC）, 20μg HepB derived in Chinese hamster ovary cell （HepB-CHO）, 10μg HepB-SC and 10 μg HepB derived in Hansenula polymorpha （HepB-HP）. Blood samples were collected one month after the third dose of primary immunization and tested for anti-HBs using chemiluminescence microparticle immunoassay （CMIA）. During the follow-up to normal and high-responders, the following information was collected： the demographic characteristic （including age and gender）, histories of hepatitis B infection, hepatitis B vaccination, smoking, drinking and chronic diseases. Blood samples were collected one month （T1） and three years after primary vaccination （T2） and anti-HBs, anti-HBc and HBsAg （if anti-HBs〈10 mU/ml） were detected by CMIA. The risk factors associated with positive rate of anti-HBs and GMC of anti-HBs were identified by multiple logistic regression analysis and multifactor linear regression model analysis, respectively. Results A total of 4 677 normal and high-responders were identified. Among 4 677 participants, 2 014 （43.06%） were males and 2 663 （56.94%） were females. The positive rate was 100%at T1 and it decreased to 80.99% （3 788/4 677） three years after vaccination. The corresponding GMC was decreased from 1 413.48 （95%CI：1 358.86-1 470.30） mU/ml to 60.33 （95%CI：56.97-63.90） mU/ml. When comparing with those vaccinated 20 μg HepB-CHO, the significantly lower positive rate of anti-HBs three years after vaccination was observed in those vaccinated 20 μg HepB-SC, 10 μg HepB-SC and 10 μg HepB-HP. The OR （95%CI） was 0.65 （0.50-0.84）, 0.52 （0.41-0.67） and 0.31 （0.28-0.45）, respectively. The GMC of anti-HBs was also significantly lower among those vaccinated 20μg HepB-SC, 10μg HepB-SC and 10 μg HepB-HP. The b （95%CI） was -0.33 （-0.47- -0.20）, -0.41 （-0.55- -0.28） and -0.78 （-0.92--0.65）, respectively. The GMC of anti-HBs in those aged 30-39 years old and 40-49 years old were lower than that in 18-29 years. The b （95%CI） was-0.31 （-0.47--0.15） and-0.24 （-0.39--0.09）, respectively. When comparing with those whose anti-HBs titer was less than 999 mU/ml at T1, the significantly higher positive rate of anti-HBs three years after vaccination was observed in those whose anti-HBs titer was 1 000-1 999 mU/ml, those whose anti-HBs titer was 2 000-9 999 mU/ml and those whose anti-HBs titer was ≥10 000 mU/ml. The OR （95%CI） was 4.97 （3.80-6.49）, 7.87 （16.19-10.01） and 9.67 （6.47-14.44）, respectively. When comparing with those whose anti-HBs titer was ≤999 mU/ml at T1, the GMC of anti-HBs three years after vaccination was also significantly higher among those whose anti-HBs titer at T1 was 1 000-1 999 mU/ml, those whose anti-HBs titer at T1 was 2 000-2 999 mU/ml and those whose anti-HBs titer at T1 was≥10 000 mU/ml. The b （95%CI） was 1.00 （0.87-1.14）, 1.85 （1.74-1.97） and 3.28 （3.12-3.44）, respectively. Four subjects showed HBsAg seroconversion and anti-HBc conversion rate was 4.68% at T2. Conclusions Anti-HBs GMC decreased rapidly three years after primary vaccination among normal and high-responder adults, while the positive rate of anti-HBs still kept at a high level. The anti-HBs persistence after primary vaccination was associated with HepB type, age and GMC of anti-HBs one month after vaccination.