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研究TBK-1基因免疫对HBcAg核酸疫苗诱导小鼠特异性体液免疫及细胞免疫的影响。采用pcDNA3.1(+)作为载体,构建真核表达质粒pcTBK-1;用HBcAg核酸疫苗(pc HBc)单独或联合质粒pcTBK-1免疫BALB/c小鼠,分别在第0、3、6周进行免疫,在第8周检测特异性抗体,脾脏淋巴细胞增殖,细胞因子IFN-γ及IL-4表达水平以及HBcAg特异性CTL活性等免疫学指标。结果显示,pc HBc联合pcTBK-1免疫小鼠的抗-HBc水平显著增高(P<0.05);淋巴细胞在体外经HBcAg刺激后,其增殖程度明显高于pc HBc单独免疫组(P<0.05);经HBcAg刺激后的淋巴细胞培养上清中,联合免疫组IFN-γ表达水平显著增高(P<0.05),而IL-4表达水平无明显改变(P>0.05);单独pc HBc免疫及联合免疫的小鼠中,HBcAg特异性CTL杀伤率均高于对照组,其中联合免疫组小鼠的CTL杀伤率显著增高(P<0.05)。提示TBK-1能够增强HBcAg核酸疫苗诱导的特异性体液免疫及细胞免疫应答,主要通过Th1型免疫应答途径,为HBV DNA疫苗的研究奠定了基础。
To investigate the effect of TBK-1 gene immunization on specific humoral and cellular immunity induced by HBcAg DNA vaccine in mice. The pcDNA3.1 (+) vector was used to construct the eukaryotic expression plasmid pcTBK-1. BALB / c mice were immunized with HBcAg nucleic acid vaccine (pc HBc) alone or in combination with pcTBK-1 plasmid at week 0, At the 8th week, immunological indexes such as specific antibody, spleen lymphocyte proliferation, cytokine IFN-γ and IL-4 expression level and HBcAg-specific CTL activity were detected. The results showed that the levels of anti-HBc in pc HBc and pcTBK-1 immunized mice were significantly increased (P <0.05). The proliferation of lymphocytes was significantly higher than that in pc HBc immunized mice (P <0.05) (P <0.05), while the expression level of IL-4 had no significant change (P> 0.05). In addition, pcBc immunization and combination Immunized mice, HBcAg-specific CTL killing rate were higher than the control group, in which the combination of immunized mice CTL killing rate was significantly increased (P <0.05). It is suggested that TBK-1 can enhance the specific humoral and cellular immune responses induced by HBcAg DNA vaccine, which lays the foundation for the study of HBV DNA vaccine through the Th1-type immune response pathway.