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Position: Home > Articles > Evaluation of Immune Effect of Clostridium Chauvoei NanA Gene Nucleic Acid Vaccine Progress in Veterinary Medicine 2019 (12) 68-73

气肿疽梭菌NanA基因核酸疫苗的免疫效果评价

作者：

陈竞;张皓;马玉腾;于成东;齐强;金鑫

单位：

延边大学珲春校区;延边大学农学院

关键词：

气肿疽梭菌;NanA基因;核酸疫苗;免疫原性

摘要：

为了检测气肿疽梭菌NanA基因核酸疫苗的免疫效果,在成功构建pVAX1-NanA真核表达重组质粒的基础上将其转染至Vero细胞,通过间接免疫荧光鉴定其表达,并将成功构建的pVAX1-NanA质粒大量提取后免疫Balb/c小鼠,设置pVAX1-NanA核酸疫苗组、pVAX1空载体组和PBS对照组。对小鼠采血并分离血清,用ELISA测定血清中的IgG、IgG1、IgG2a抗体水平和IFN-γ、IL-4细胞因子水平;第3次免疫2周后,用流式细胞仪测定小鼠脾脏中的CD4~+和CD8~+T细胞水平。结果表明,pVAX1-NanA核酸疫苗免疫Balb/c小鼠后,小鼠血清中IgG、IgG1、IgG2a的抗体水平极显著高于pVAX1空载体组和PBS对照组(P<0.01);IFN-γ和IL-4细胞因子水平极显著高于pVAX1空载体组和PBS对照组(P<0.01);CD4~+、CD8~+T细胞分析表明, pVAX1-NanA免疫组的CD4~+和CD8~+T细胞水平显著高于pVAX1组和PBS对照组(P<0.05),pVAX1-NanA核酸疫苗组的CD4~+/CD8~+T细胞比值高于pVAX1空载体组与PBS对照组(P<0.05),pVAX1空载体组和PBS对照组之间无显著差异(P>0.05)。试验成功制备了pVAX1-NanA核酸疫苗,且pVAX1-NanA核酸疫苗可刺激Balb/c小鼠产生体液免疫和细胞免疫。

译名：

Evaluation of Immune Effect of Clostridium Chauvoei NanA Gene Nucleic Acid Vaccine

作者：

CHEN Jing;ZHANG Hao;MA Yu-teng;YU Cheng-dong;QI Qiang;JIN Xin;College of Agriculture,Yanbian University;College of Yanbian University Hunchun Campus;

关键词：

Clostridium chauvoei;;NanA gene;;nucleic acid vaccine;;immunogenicity

摘要：

In order to detect the immune effect of Clostridium chauvoei NanA gene nucleic acid vaccine,this experiment successfully built the eukaryotic expression recombinant plasmid pVAX1-NanA,and transfected into Vero cells,the expression was identified by indirect immunofluorescence,after the pVAX1-NanA plasmid was extracted,Balb/c mice were immunized,pVAX1-NanA nucleic acid vaccine group,pVAX1 empty vector group and PBS control group were set.The sera of mice were collected,and IgG,IgG1,IgG2a antibody levels and IFN-γ,IL-4 were measured by ELISA.After 2 weeks of the third immunization,CD4~+ and CD8~+ levels in mouse spleen lymphocytes were measured by flow cytometry.The results showed that after immunization of Balb/c mice with pVAX1-NanA nucleic acid vaccine,the antibody levels of IgG,IgG1 and IgG2a in sera of mice were significantly higher than those of pVAX1 empty vector group and PBS control group(P<0.01); IFN-γ and IL-4 cytokine levels in sera of mice were significantly higher than that of pVAX1 empty vector group and PBS control group(P<0.01). lymphocyte subsets analysis showed significantly higher levels of CD4~+ and CD8~+ T cells in the pVAX1-NanA immunized group,compared with the pVAX1 group and the PBS control group(P<0.05),the ratio of CD4~+/CD8~+ in the pVAX1-NanA nucleic acid vaccine group was higher than that in the pVAX1 empty vector group and the PBS control group(P<0.05),and there was no significant difference between the pVAX1 empty vector group and the PBS control group(P>0.05).The pVAX1-NanA nucleic acid vaccine was successfully prepared,and the pVAX1-NanA nucleic acid vaccine stimulated humoral and cellular immunity in Balb/c mice

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气肿疽梭菌NanA基因核酸疫苗的免疫效果评价

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