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目的 研究HBsAg 抗HBs 质粒DNA复合型疫苗的免疫原性及其诱生细胞免疫应答的类型。方法 分别以HBsAg、HBsAg 抗 HBs(IC)、pI/AmpHBs、IC pI/AmpHBs及IC pI/Amp免疫小鼠 ,检测抗 HBs的效价 ,分析抗 HBsIgG亚类 (ELISA) ;取免疫小鼠脾细胞 ,体外抗原刺激 ,用竞争性RT PCR方法检测IFN γ及IL 4mRNA转录水平。结果 IC pI/AmpHBs诱生的抗 HBs效价明显高于IC或pI/AmpHBs单独免疫组 ,其IgG2a/IgG1比值高于IC免疫组 ,而低于pI/AmpHBs免疫组。IC pI/AmpHBs免疫小鼠脾细胞在HBsAg刺激下 ,IFN γmRNA转录水平明显高于其他免疫组 ,其IFN γmRNA的T/C(目的片段Target/竞争片段Competitor)比值为IC免疫小鼠脾细胞的 10倍 ;IC pI/AmpHBs免疫小鼠脾细胞IL 4mRNA转录水平亦高于其他免疫组 ,其IL 4mRNA的T/C比值为IC免疫小鼠脾细胞的 2倍。结论 HBsAg 抗HBs 质粒DNA复合型疫苗在增强体液免疫应答的同时可诱导脾细胞IFN γ的表达水平增高。
Objective To study the immunogenicity of HBsAg anti-HBs plasmid DNA composite vaccine and its type of induced cellular immune response. Methods The mice were immunized with HBsAg, HBsAg anti-HBs (IC), pI / AmpHBs, IC pI / AmpHBs and IC pI / Amp respectively to detect the anti-HBs titer and analyze anti-HBs IgG subclass Cells and in vitro antigens were stimulated. IFNγ and IL 4 mRNA transcription levels were detected by competitive RT-PCR. Results The titer of anti-HBs induced by IC pI / AmpHBs was significantly higher than that of IC or pI / AmpHBs alone group, and the ratio of IgG2a / IgG1 was higher than that of IC and lower than that of pI / AmpHBs. The transcription level of IFN-γmRNA of splenocytes immunized with IC pI / AmpHBs was significantly higher than that of other immunized groups under the stimulation of HBsAg. The ratio of T / C of IFNγmRNA (Target / Competitor) was 10-fold. The transcription level of IL-4 mRNA in splenocytes of ICI / AmpHBs-immunized mice was also higher than that of other immunized groups. The T / C ratio of IL-4mRNA was 2-fold that of splenocytes in IC-immunized mice. Conclusion HBsAg anti-HBs plasmid DNA composite vaccine can enhance the humoral immune response and induce IFNγ expression in splenocytes.