摘要
目的以300L发酵罐中试规模生产的冻干Mtb Ag85A质粒DNA疫苗为基础免疫小鼠,对该疫苗的免疫效能进行研究。方法将4～6周龄的BALB/c小鼠20只通过数字表法随机分为2组,一组免疫MtbAg85A质粒DNA疫苗(结核DNA疫苗免疫组),另一组免疫PBS作为阴性对照(PBS对照组);于免疫前采血,肌内注射免疫3次。在第3次免疫后3周摘眼球处死小鼠,收集血清,进行血清特异性抗体检测和细胞因子(IFN-γ、IL-4)检测;同时对每只小鼠取脾细胞进行特异性γ干扰素(IFN-γ)分泌检测和脾细胞CD4+、CD8+百分率检测。结果 (1)免疫前后免疫组与对照组抗体水平吸光度A值均小于0.2。(2)结核DNA疫苗免疫组血清细胞因子IL-4水平[(146.2±34.3)pg/ml]低于PBS对照组[(177.7±28.1)pg/ml],差异有统计学意义(t=2.244,P=0.038),而IFN-γ水平[(129.6±159.0)pg/ml]虽然高于PBS对照组[(76.5±21.5)pg/ml],但差异无统计学意义(t=-1.047,P=0.309)。(3)酶联免疫斑点检测结果表明结核DNA疫苗免疫组脾细胞特异性IFN-γ分泌水平(分泌IFN-γ细胞个数)(103.60±112.14)高于PBS对照组(5.78±5.83),差异有统计学意义(t=36.538,P=0.018)。(4)结核DNA疫苗免疫组CD4+细胞百分率均值(21.57%)高于PBS对照组(12.17%),差异有统计学意义(t=3.043,P=0.038);CD8+细胞百分率均值(13.70%)与PBS对照组(10.57%)相比,差异无统计学意义(t=0.847,P=0.445)。结论Mtb Ag85A质粒DNA疫苗主要刺激机体Th1型细胞免疫,而对诱导机体产生Th2型细胞免疫应答没有显著的促进作用。
Objective To study the immunogenicity of Mycobacterial tuberculosis Ag85A DNA vaccine with pilot scale production in mice.Methods Twenty BALB/c mice were randomly divided into 2 groups,and then immunized intramuscularly for 3 times with Mtb Ag85A DNA or PBS respectively.The mice were killed and the whole blood samples were collected at 3 weeks after the last immunization.The specific antibody and cytokines（IL-4） in sera were detected.The Mtb Ag85A specific interferon-γ（IFN-γ） level and the percentages of CD4＋ or CD8＋ T cell in mouse spleen cells were also detected.Results The data obtained by statistical analyses demonstrated that：（1） The absorbance values of Ag85A-specific antibody in the sera of each group mice before and after immunization were all less than 0.2.（2） The IL-4 level in sera from Ag85A DNA group [（146.2±34.3） pg/ml] was significantly lower than that from PBS group [（177.7±28.1） pg/ml]（t=2.244,P=0.038）.The IFN-γ level in sera from Ag85A DNA group [（129.6±159.0） pg/ml]was higher than that from PBS group [（76.5±21.5） pg/ml],but there was no significant difference（t=-1.047,P=0.309）.（3） The data from the ELISPOT assay for IFN-γ showed that the number of spot-forming spleen cells in Ag85A DNA group（103.60±112.14） was significantly higher than that in PBS group（5.78±5.83）（t=36.538,P=0.018）.（4） The percentage（21.57%） of CD4＋ T cells in Ag85A DNA group was higher than that（12.17%） in PBS group（t=3.043,P=0.038）;however,there was no significant difference on the percentage of CD8＋ T cells between the two groups（13.70% vs.10.57%,t=0.847,P=0.445）.Conclusion The Th1 cellular immunity was mainly stimulated in the mice immunized by Mtb Ag85A DNA vaccine.But Th2 cellular immune response was not significantly induced in these mice.
出处
《中国防痨杂志》
CAS
2012年第3期168-171,共4页
Chinese Journal of Antituberculosis
基金
"十一五"国家科技重大专项(2008ZX-10003-013(2))
