结核分枝杆菌三基因融合表达的亚单位疫苗研究

发布时间：2018-03-31 09:14

本文选题：结核分枝杆菌　切入点：亚单位疫苗　出处：《中国预防兽医学报》2017年11期

【摘要】：为筛选出安全、有效的抗结核分枝杆菌感染的亚单位疫苗,本研究选择了9个结核分枝杆菌的重要抗原(Rv0199、Rv0394c、Omp A、Rv3802c、Ag85a、TB10.4、Rv2660c、Rv0125、Mpt51),利用小鼠模型评价其免疫效果。每3个抗原一组串联融合表达,将其分别与弗氏不完全佐剂及沙门氏菌菌蜕混合,共分成7组,通过皮下注射的方式免疫小鼠,共免疫3次。每次免疫后两周检测其血清抗体分型与效价、脾脏淋巴细胞的分化及其细胞因子的分泌。结果表明,这些融合蛋白均能够刺激机体产生高滴度的抗体,具有较强的免疫原性,其中融合蛋白Rv0199-Rv0394c-Omp A在一免后2周即可以刺激机体产生较高滴度的抗体。抗体分型结果显示,除阴性对照组外,其余组小鼠血清中Ig G1的滴度均高于Ig G2a。并且机体的免疫反应倾向于体液免疫。其中,沙门氏菌菌蜕的佐剂作用与商品化的弗氏不完全佐剂免疫效果一致。本研究为研制结核分枝杆菌亚单位疫苗奠定了基础。
[Abstract]:In order to screen out a safe and effective subunit vaccine against Mycobacterium tuberculosis infection, we selected 9 important antigens of Mycobacterium tuberculosis, Rv0199994, Rv0394c, omp Agno Rv3802cngAg85a1, TB10.4, Rv2660cU, Rv0125Mpt51. the immunological effect of the vaccine was evaluated by using a mouse model. Each group of three antigens was fused in tandem. They were mixed with Freund's incomplete adjuvant and salmonella slough, and were divided into 7 groups. The mice were immunized by subcutaneous injection for 3 times. The serum antibody typing and titer were detected two weeks after each immunization. The differentiation of spleen lymphocytes and the secretion of cytokines. The results show that these fusion proteins can stimulate the production of high titers of antibodies and have strong immunogenicity. The fusion protein Rv0199-Rv0394c-Omp A could stimulate the body to produce higher titer antibodies 2 weeks after immunization. The results of antibody typing showed that, except for negative control group, The titer of IgG G1 in serum of the other groups was higher than that of IgG2a, and the immune response of the body tended to be humoral immunity. The adjuvant effect of salmonella slough was consistent with that of commercial Freund's incomplete adjuvant. This study laid a foundation for the development of Mycobacterium tuberculosis subunit vaccine.
【作者单位】：中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物细菌病创新团队;
【基金】：国家自然科学基金面上项目(31272538) 兽医生物技术国家重点实验室基本科研业务费项目(SKLVBP2015006) 牛羊常见细菌病的诊断与检测新技术研究/牛羊副结核分枝杆菌诊断与检测新技术研究(2016YFD0500905)
【分类号】：R392

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