幽门螺杆菌表面抗原免疫保护作用的体外与活体研究

目的：调查幽门螺杆菌（Hp)几种表面蛋白体外对T细胞增殖的影响和在小鼠体内的免疫保护作用。方法：评价Hp全菌抗原、尿原酶（Urease)、黏附素（hpaA)、外膜蛋白25（Hop25)和38（Hop38)对人外周血T细胞及小鼠CD4^ T细胞增殖的影响；与佐剂合用，评价上述重组蛋白对小鼠Hp感染的免疫预防作用。结果：Urease和Hop25可刺激人及鼠T细胞增殖，hapA只能刺激Hp^ PBL增殖，而Hop38则有毒性作用；Hop25和Hop38均可产生60％的完全保护，hpaA可产生100％的部分保护即降低细菌定植密度，而Urease只能产生40％的部分保护。结论：外膜蛋白可能是一组高效的Hp疫苗免疫原；其长期免疫效果及对T细胞功能的活体调节作用尚需进一步评价。国际上尚未见相关报道。

THE IN VIVO AND IN VITRO EVALUATION OF THE IMMUNE PROTECTIVE ACTIVITY OF THE RECOMBINANT SURFACE PROTEINS OF HELICOBACTER PYLORI

Objective:To evaluate the effect of several surface proteins of Helicobacter pylori(H pylori) on the proliferation of peripheral blood lymphocytes(PBL) and CD 4 + spleenocytes of mice and to determine their effect to protect H pylori infection in mice via immunization.Methods:PBL of H pylori infectious and unifectious patients were isolated and CD 4 + spleenocytes were isolated from mice.Their proliferation in response to the surface proteins of H pylori such as recombinant urease,hpaA,and outmembrane protein(Hop)25 and 38 were determined by the 3 H thymidine incorporation assay.Moreover,The colonization of H pylori in mice was evaluated after the immunization with these proteins as well as mucosal adjuvant,cholera toxin.Results:Urease and Hop25 were capable of inducing T cell proliferation,while hpaA can only induce the proliferation of T cell isolated from H pylori infectious subjects.The proliferation of both human and mouse T cells was inhibitied by Hop38.In regard to the immunization,Hop25 and 38 protected 60% fo the mice from infection.hpaA decreased the colonization of H pylori in all of the animals,whereas urease only generated 40% of partial protection.Conclusion:Outmenbrane protein of H pylori seems to be good immunogen candidates in the vaccine preparation.The long term effect of immunization and their effect in regulating T cell function in vivo remain to be studied.