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目的 制备表达幽门螺杆菌 (H pylori)尿素酶B亚单位 (UreB)的减毒鼠伤寒杆菌活菌重组疫苗 ,并观测Balb/c小鼠口服该重组疫苗后的免疫效果。方法 构建表达UreB的重组减毒鼠伤寒杆菌的疫苗株 ,SL3 2 61 /pTC0 1 UreB ,并用Western blot检测UreB的表达。将SL3 2 61 /pTC0 1 UreB口服免疫Balb/c小鼠 ,1 2周后检测肠液和血清中的特异性抗体反应及脾细胞培养液上清中的IFN γ和IL 1 0含量。结果 Western blot显示减毒鼠伤寒杆菌株SL3 2 61 /pTC0 1 UreB能表达约 61KD的蛋白 ,与H pyloriUreB亚单位蛋白分子量相符 ,具有抗原性。口服免疫小鼠后 ,疫苗组小鼠的肠液和血清中可分别检测到针对UreB的特异性IgA和IgG抗体 ,小鼠脾细胞培养上清中的IFN γ和IL 1 0的含量亦明显升高。病理学检查显示疫苗组小鼠与对照组小鼠胃粘膜炎症指数无差异。结论 表达H pyloriUreB的减毒鼠伤寒杆菌SL3 2 61 /pTC0 1 UreB可用作抗H pylori感染的口服疫苗
Objective To prepare an attenuated attenuated Salmonella typhimurium recombinant vaccine expressing the H pylori urease B subunit (UreB) and observe the immune effect of the recombinant vaccine administered to Balb / c mice. Methods The recombinant attenuated Salmonella typhimurium vaccine strain expressing UreB was constructed, SL3 2 61 / pTC0 1 UreB, and the expression of UreB was detected by Western blot. Balb / c mice were orally immunized with SL3 2 61 / pTC0 1 UreB and the specific antibody responses in intestinal juice and serum and the contents of IFN γ and IL 1 0 in the supernatants of splenocytes were detected after 1 and 2 weeks. Results Western blot showed that the attenuated Salmonella typhimurium strain SL3 2 61 / pTC0 1 UreB expressed about 61 kD protein, which was consistent with the molecular weight of H pylori UreB subunit protein and was antigenic. After oral immunization, the specific IgA and IgG antibodies against UreB were detected in the intestinal fluid and serum of mice in the vaccine group, and the levels of IFN-γ and IL-10 in the spleen cell culture supernatant of mice were also significantly increased . Pathological examination showed no difference in gastric mucosal inflammatory index between vaccine group and control group. Conclusion The attenuated Salmonella typhimurium SL3 2 61 / pTC0 1 UreB expressing H pylori UreB can be used as an oral vaccine against H pylori infection