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探讨表达幽门螺杆菌 (Helicobacterpylori,Hp)粘附素保守区 (AB)的减毒鼠伤寒沙门菌X4 0 72 (pYA2 4 8 AB)的免疫治疗效果 ,以及可能的免疫治疗机制 ,以确定X4 0 72 (pYA2 4 8 AB)在Hp疫苗研制中的应用价值 .建立Hp感染小鼠模型 ,在该模型中评价X4 0 72 (pYA2 4 8 AB)治疗Hp感染的效果 ,运用细菌培养法观察Hp根除率及定植量的改变 ,流式细胞术分析免疫后小鼠脾细胞亚型 ,IL 2和IL 4细胞依赖株的细胞测活法检测细胞因子 ,ELISA法检测小鼠血清及肠液中抗AB抗体产生情况 .结果表明 ,免疫治疗后疫苗治疗组根除率为 5 3 3% ,显著高于X4 0 72 (pYA2 4 8)和PBS对照组 (P <0 0 1 ) .未根除Hp的小鼠 ,疫苗治疗组Hp的定植密度明显低于其他两组 (P <0 0 1 ) .应用流式细胞仪分析免疫小鼠脾CD4 + /CD8+ T淋巴细胞的比值时 ,发现疫苗治疗组和鼠伤寒菌对照组的比值均显著高于PBS对照组 (P <0 0 5 ) ,而疫苗治疗组的比值又显著高于鼠伤寒菌对照组 (P <0 0 5 ) .进一步对细胞因子进行分析发现 ,与PBS对照组相比免疫治疗组和鼠伤寒菌对照组IL 2和IL 4明显升高(P <0 0 5 ) .同时 ,免疫治疗组与鼠伤寒菌对照组相比 ,IL 4增高明显 (P <0 0 5 ) .针对AB的抗体测定结果发现 ,仅在免疫治疗组的小鼠肠液中检测到了IgA抗体
To investigate the immunotherapeutic effect of attenuated Salmonella typhimurium X4 072 (pYA2 4 8 AB) expressing Helicobacter pylori (Hp) adhesin conserved region (AB) and possible immunotherapeutic mechanisms to determine X4 0 72 (pYA2 4 8 AB) in the development of Hp vaccine.A mouse model of Hp infection was established and the effect of X4 0 72 (pYA2 4 8 AB) on Hp infection was evaluated in this model. The bacterial culture method was used to observe the eradication of Hp Rate and colonization rate. Cytokines were detected by flow cytometry (FCM) in subtypes of splenocytes after immunization, IL-2 and IL-4 cell-dependent cell viability assay, anti-AB antibody in serum and intestinal juice The results showed that the eradication rate of vaccine treatment group after immunotherapy was 53.3%, which was significantly higher than that of X4 0 72 (pYA2 48) and PBS control group (P 0 01) .The mice that did not eradicate Hp, The colonization densities of Hp in the vaccine-treated group were significantly lower than those in the other two groups (P <0.01). When the ratio of splenic CD4 + / CD8 + T lymphocytes in immunized mice was analyzed by flow cytometry, it was found that the vaccine treated group and S. typhimurium The ratio of the control group were significantly higher than the PBS control group (P <0 05), while the vaccine treatment group (P <0.05) .Further analysis of cytokines showed that compared with the PBS control group, the levels of IL-2 and IL-4 in the immunotherapy group and the control group were significantly increased (P < P <0.05) .At the same time, IL 4 increased significantly in the immunotherapy group compared with the control group (P <0.05) .Ab antibody assay results showed that only in the immunotherapy group of mouse intestinal fluid IgA antibody was detected