鼠疫双组分疫苗的免疫学评价

发布时间：2018-06-19 10:46

本文选题：F1+rV + 蛋白　；参考：《福建医科大学》2011年硕士论文

【摘要】：目的:本课题以提取的鼠疫F1抗原和重组鼠疫V抗原免疫小鼠,豚鼠两种动物模型,评价鼠疫亚单位疫苗的免疫效果,为鼠疫疫苗研制奠定基础。方法:1.在小鼠动物模型上,本课题用F1和rV与氢氧化铝进行物理混合后免疫小鼠。通过脾淋巴细胞增殖实验、脾淋巴细胞细胞因子,流式细胞分析术和抗体滴度的测定四种方法对F1和rV蛋白疫苗的免疫原性进行了初步评价。2.在豚鼠动物模型上,实验采用不同剂量的F1和rV抗原与氢氧化铝佐剂进行物理混合后免疫豚鼠,以探求合适的疫苗浓度;通过分离免疫后不同时间的外周血单个核细胞进行增殖实验,对疫苗末次注射八周后的豚鼠进行过敏性迟发型超敏反应实验,并测定免疫后不同时间抗体滴度等实验,对F1和rV蛋白疫苗的免疫原性进行了评价。3.在疫苗效力保护实验中,通过不同剂量的鼠疫双组分疫苗免疫小鼠,并于一周后用一定剂量的鼠疫菌株对实验小鼠进行攻击,选择一定的时间对小鼠脾脏的细菌进行培养计数,从而观察疫苗的保护效果。结果:1.小鼠动物模型结果:ELISA检测结果显示F1+rV+Al(OH)3组有较强的体液免疫应答;ELISPOT,MTT细胞增殖结果显示F1+rV+Al(OH)3组在不同时间点经F1,rV抗原刺激后分泌IFN-r的脾脏淋巴细胞数均高于其余各组。2.豚鼠动物模型结果:ELISA检测结果显示各疫苗组均有较强的体液免疫应答,同时活疫苗组对F1抗原特异性的体液IgG效价明显高于其余各组,结果具有统计学意义。MTT细胞增殖结果显示不同剂量的疫苗组F1+rV组及活疫苗组在不同时间点经F1,rV抗原刺激后的增殖与阴性对照组无显著性差异;迟发型超敏反应显示F1+rV复合疫苗能介导较强的细胞免疫应答。3.疫苗效力保护实验结果显示阴性对照组有数量不等的菌落生长,而疫苗组小鼠脾脏均无细菌生长。结论:鼠疫双组分疫苗能引起小鼠和豚鼠明显的免疫应答,Al(OH)3佐剂能够显著提高其应答水平,同时对小鼠具有一定的保护力。确证了其与氢氧化铝佐剂物理混合后能够激发高水平的免疫应答,为进一步研制新型鼠疫疫苗奠定了基础
[Abstract]:Objective: to evaluate the immune effect of Yersinia pestis subunit vaccine in mice and guinea pigs by immunizing mice with F1 antigen and recombinant V antigen in order to lay a foundation for the preparation of Yersinia pestis vaccine. Method 1: 1. In this study, mice were immunized with F1 and rV and aluminum hydroxide. The immunogenicity of F1 and rV protein vaccines was preliminarily evaluated by spleen lymphocyte proliferation test, spleen lymphocyte factor, flow cytometry and antibody titer determination. The guinea pigs were immunized with different doses of F1 and rV antigens and aluminum hydroxide adjuvant in order to find the appropriate vaccine concentration. The proliferation of peripheral blood mononuclear cells (PBMC) at different time after immunization was carried out, the allergic delayed type hypersensitivity test was performed on guinea pigs after the last injection of the vaccine eight weeks later, and the antibody titers were measured at different times after immunization. The immunogenicity of F1 and RV protein vaccines was evaluated. In the protective experiment of vaccine efficacy, mice were immunized with different doses of plague two-component vaccine, and a certain dose of Yersinia pestis strain was used to attack the experimental mice one week later, and selected a certain time to culture and count the bacteria in the spleen of mice. The protective effect of the vaccine was observed. The result is 1: 1. The results of mouse animal model showed that F1 rV Albumin OHH3 group had a strong humoral immune response and ELISPOTH MTT cell proliferation. The results showed that the spleen lymphocytes secreting IFN-r in F1 rV Alo OHH3 group were higher than those in other groups at different time points after stimulation by F1 rV antigen. The results of the guinea pig animal model showed that each vaccine group had a strong humoral immune response, and the live vaccine group had higher specific humoral IgG titer against F1 antigen than the other groups. Results the results of MTT cell proliferation showed that there was no significant difference between F1 rV group and live vaccine group stimulated by F1rV antigen at different time points. Delayed type hypersensitivity reaction showed that F1 rV complex vaccine could mediate strong cellular immune response. 3. The results of vaccine protection test showed that there were different numbers of colonies in negative control group, but no bacteria growth in spleen of vaccine group. Conclusion: Yersinia pestis two-component vaccine can induce a significant immune response in mice and guinea pigs. It was confirmed that it could stimulate a high level of immune response after physical mixing with aluminum hydroxide adjuvant, which laid a foundation for the further development of a new plague vaccine.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392

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