免疫复合物型乙型肝炎疫苗的免疫原性分析

摘要

目的：分析免疫复合物型乙型肝炎疫苗在小鼠体内和食蟹猴体内的免疫原性。方法免疫复合物型乙型肝炎疫苗单次免疫小鼠，不同时间点采血进行anti-HBs滴度动态检测；不同免疫程序免疫小鼠，进行anti-HBs滴度检测；肌肉注射5μg/只免疫小鼠，ELISPOT方法检测IFN-γ形成斑点数并进行阳转率统计；测定稳定性试验样品的ED50；分别以20μg/只和100μg/只剂量免疫食蟹猴，不同时间点采血检测anti-HBs滴度。结果免疫复合物型乙型肝炎疫苗动态检测anti-HBs滴度均高于重组乙型肝炎疫苗（酿酒酵母）；免疫复合物型乙型肝炎免疫2针次anti-HBs滴度可达到重组乙型肝炎疫苗（酿酒酵母）免疫3针次水平；免疫复合物型乙型肝炎疫苗细胞免疫检测IFN-γ斑点形成细胞（ SFC）及阳转率高于重组乙型肝炎疫苗（酿酒酵母）及酿酒乙型肝炎疫苗国家参考品；免疫复合物型乙型肝炎疫苗长期稳定性优于酿酒酵母乙肝疫苗，ED50值低于后者，高低两种剂量免疫食蟹猴，均可诱导高滴度anti-HBs，且持续性良好。结论免疫复合物型乙型肝炎疫苗有较好的免疫原性，其体液免疫和细胞免疫水平均优于重组乙型肝炎疫苗（酿酒酵母）。%Objective To evaluate the immunogenicity of an immune complexed hepatitis B vac-cine ( HBsAg-HBIG immune complexes , IC) in mouse and cynomolgus monkeys by using recombinant hepa-titis B vaccine ( Saccharomyces cerevisiae, HBsAg) as the control .Methods BALB/c mice were vaccinated with single dose of IC and single dose of HBsAg respectively and then serum samples were collected at differ -ent time points for the detection of dynamic anti-HBs by using ELISA .The serum anti-HBs titers in BALB/c mice vaccinated with different immunization strategies were also analyzed .ELISPOT assay was performed to detect the numbers of IFN-γSFC and IFN-γpositive rate in splenocytes of BALB/c mice intramuscularly im-munized with IC, HBsAg or standard hepatitis B vaccine at 5μg/mouse.ED50 was measured to evaluate the stability of IC.Twelve cynomolgus monkeys were equally divided into two groups and immunized with high dose (100 μg) and low dose (20 μg) of IC respectively and then , serum anti-HBs levels at different time points were detected .Results The serum anti-HBs titers in IC immunized group at different time points were higher than those immunized with HBsAg .Moreover, the anti-HBs titer induced by two doses of IC reached a level comparable to that elicited by three doses of HBsAg .ELISPOT assay showed that both the numbers of IFN-γSFC and IFN-γpositive rate were the highest in IC immunized group as compared with those immunized with HBsAg and standard hepatitis B vaccine .IC had a lower ED50 than HBsAg, indicating a good long term stability .Cynomolgus monkeys immunized with high or low dose of IC produced high levels of anti-HBs titer during a long time period .Conclusion IC has a higher immunogenicity inducing both hu-moral immunity and cellular immunity as compared with HBsAg or standard hepatitis B vaccine .