摘要
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 adjuvant effect of TBK-1 on the induction of humoral and cellular immune responses by HBV core DNA vaccine,the eukaryotic expression plasmid pcTBK-1 was constructed by inserting mouse TBK-1 gene into the vector pcDNA3.1（＋）,and BALB/c mice were immunized by three intramuscular injections of HBcAg DNA vaccine plasmid alone or in combination with pcTBK-1 at 0 th,3 th and 6 th week.At 8 th week,the anti-HBc level,splenocyte proliferation,IFN-γ and IL-4 level and HBcAg specific CTL activity were measured.It was found that the level of anti-HBc in the co-immunization group was higher than that in group of mice immunized with HBV core DNA vaccine alone（P0.05）.In addition,the HBcAg specific splenocyte proliferation in the co-immunization group was significantly higher than that in the pcHBc single immunization group（P0.05）.The level of IFN-γ in the supernatant of splenocyte cultured with HBcAg in vitro in the co-immunization group was significantly increased（P0.05） while the level of IL-4 showed no significant difference（P0.05）.Also,the CTL activities both in the pcHBc group and co-immunization group were increased,while the CTL activity in co-immunization groupwasincreased more significantly than that in pcHBc group（P0.05）.Thus,the plasmid encoding TBK-1 together with HBcAg DNA vaccine can enhance the humoral and cellular immune responses induced in mice mainly through the Th1-like way,and this study can lay a foundation for the further study on the HBV DNA vaccines.
出处
《现代免疫学》
CAS
CSCD
北大核心
2010年第6期443-447,共5页
Current Immunology
基金
浙江省自然科学基金资助项目(Y207534)
