气肿疽梭菌pVAX1-NagH 基因核酸疫苗的免疫效果评价  

Evaluation of immune effect of nucleic acid vaccine of Clostridium chauvoei pVAX1-NagH

作　　者：曲庭伟 张文璇 崔春梅 姜人月 鲍君铎 赵仁双 金鑫[1] QU Tingwei;ZHANG Wenxuan;CUI Chunmei;JIANG Renyue;BAO Junduo;ZHAO Renshuang;JIN Xin(Agricultural College of Yanbian University,Yanji Jilin 133002,China;Shanghai Ruiyi Biotechnology Co.Ltd.,Shanghai 201199,China)

机构地区：[1]延边大学农学院,吉林延吉133002 [2]上海市锐翌生物科技有限公司,上海201199

出　　处：《延边大学农学学报》2022年第1期63-68,共6页Agricultural Science Journal of Yanbian University

基　　金：高等学校学科创新引智计划资助(D20034);东北寒区优质肉牛高效安全养殖技术应用与示范(2018YFD0501702)。

摘　　要：为检测气肿疽梭菌中NagH基因的核酸疫苗的免疫效果,在成功构pVAX1-NagH真核表达重组质粒后,将其转染至Vero细胞,通过间接免疫荧光方法对重组体进行鉴定。鉴定成功后,大量提取重组体质粒,对BALB/c小鼠进行免疫,免疫方法为肌肉注射。设置pVAX1-NagH核酸疫苗组、载体对照组以及PBS阴性对照组。对小鼠采血后分离血清,采用ELISA方法对IgG、IgG1、IgG2a抗体水平和IL-4、IFN-γ细胞因子水平进行测定。结果表明:IgG、IgG1、IgG2a抗体水平与IL-4、IFN-γ细胞因子水平呈显著上升趋势,且与另外2个对照组相比,均存在显著性差异,而2个对照组间未表现出显著性差异。该试验说明初步制备了pVAX1-NagH核酸疫苗,并且pVAX1-NagH核酸疫苗可刺激BALB/c小鼠产生良好的体液免疫和细胞免疫。In order to detect the immune effect of the nucleic acid vaccine of NagH gene in Clostridium chauvoei,the pVAX1-NagH eukaryotic expression recombinant plasmid was constructed successfully,then transfected it into Vero cells,and was identified by indirect immunofluorescence method.After successful identification,a large number of recombinant constitutional granules were extracted and immunized BALB/c mice by intramuscular injection.Three groups were set up,which named pVAX1-NagH nucleic acid vaccine group,vector control group and PBS negative control group.Serum from blood of the mice was isolated,the levels of IgG,IgG1 and IgG2a antibodies,IL-4 and IFN-γcytokine levels were measured by ELISA.The results showed that the IgG,IgG1,IgG2a antibody levels and IL-4,IFN-γcytokines levels had a significant upward trend.Compared with the other two control groups,there were significant differences,but there was no significant difference between the two control groups.This experiment illustrates that pVAX1-NagH nucleic acid vaccine has been prepared preliminarily,and pVAX1-NagH nucleic acid vaccine can stimulate BALB/c mice to produce humoral immunity and cellular immunity.

关 键 词：气肿疽梭菌 NagH 核酸疫苗 免疫 小鼠 

分 类 号：S852.61[农业科学—基础兽医学]