乙型肝炎表面抗原-破伤风类毒素结合疫苗的制备及免疫学特性  

Preparation and immunological properties of hepatitis B virus surface antigen-tetanus toxoid conjugate vaccine

作　　者：崔仑标[1] 胡忠玉[2] 葛以跃[1] 翟祥军[1] 史智扬[1] 焦永军[1] 戚宇华[1] 梁争论[2] 李凤祥[2] 汪华[1] 

机构地区：[1]江苏省疾病预防控制中心微生物科,南京210009 [2]中国药品生物制品检定所

出　　处：《中华微生物学和免疫学杂志》2008年第11期1016-1019,共4页Chinese Journal of Microbiology and Immunology

基　　金：江苏省科技支撑计划（BE2008683）;江苏省科技创新人才启动项目（BK2007608）

摘　　要：目的制备乙型肝炎（乙肝）表面抗原（HBsAg）与破伤风类毒素（TT）结合疫苗，并研究其免疫学特性。方法TT经溴化氢活化后与己二酰肼形成TT-酰肼基衍生物，在碳二亚胺作用下与HBsAg共价结合。将结合物、HBsAg及乙肝疫苗免疫小鼠，并设生理盐水对照组，分别于7、14、21、28d取血，应用化学发光法检测小鼠血清抗-HBs，于第7、28天应用酶联免疫斑点法（ELISPOT）检测脾单个核细胞（mononuclear cells，MNC）分泌HBsAg特异性IFN-γ和IL-2能力。结果成功制备了HBsAg—TT结合物，该结合物诱导抗-HBs阳转率及抗体滴度均较单独免疫HBsAg及乙肝疫苗组高，主要以IgG2a抗体为主，且分泌IFN-γ和IL-2的淋巴细胞数也显著增加。结论用该方法制备的HBsAg-TT结合疫苗，不仅可诱导较强的体液免疫，还可诱导以TH1应答为主的细胞免疫。Objective To prepare and study the immunogenicity of hepatitis B virus surface antigen （HBsAg）-tetanus toxoid （TT） conjugate vaccine. Methods TT was activated by cyangen bromide and reacted with adipic acid dihydrazide, then HBsAg-TT conjugate was prepared by carbodiimide. Conjugate, HBsAg or hepatitis B vaccine was injected subcutaneously into mice. Anti-HBsAg and HBsAg-specific T cell response elicited by these immunogens were assayed. Results New HBsAg-TT conjugate elicited higher levels of anti-HBsAg and HBsAg positive conversion rates after the immunization than did HBsAg alone or hepatitis B vaccine. Conjugate induced mostly antibodies of the IgG2a subclass, while HBsAg alone or hepatitis B vaccine mainly elicited anti-HBsAg in the IgG1 subclass. The number of IFN-γand IL-2 secreting T cells induced by conjugate was also significantly higher than that did by HBsAg or hepatitis B vaccine. Conclusion This study indicated new HBsAg-TT conjugate can induce both stronger humoral and TH 1 type of cellular immune response.

关 键 词：乙型肝炎表面抗原 破伤风类毒素 结合疫苗 免疫应答 

分 类 号：R392[医药卫生—免疫学]