国家高技术“863”计划资助项目(102-07-03-06),国家自然科学基金资助项目(30270078)和军队“十五”医药卫生科研课题资助项目(OIMA-132).
This work was supported by grants from The State 863 High Technology R & D Project of China(102-07-03-06),The National Natural Sciences Foundation of China(30270078) and Mlitary ‘Ten-Five’ Commanding Project.
探讨幽门螺杆菌(Hp)保守区(AB)蛋白的体外安全性、免疫原性和黏附作用,以确定AB在Hp疫苗研制中的应用价值.ELISA法测定Hp感染者血清中抗AB抗体,四唑盐比色法(MTT)测定T细胞对AB的增殖反应,流式细胞术检测AB致T细胞表达FasL的作用,二苯胺(DPA)法测定AB致T细胞凋亡率,光镜计数法研究AB抗体对Hp与胃癌细胞黏附的影响.ELISA法共检测了55份血清,以快速尿素酶实验(RUT)作为平行对照,两法的评价判断一致性程度的指标卡帕系数为0.76. 同时,低剂量AB即可刺激Hp+T细胞的增殖.体外安全性实验表明,AB无明显调节T细胞表达FasL的作用以及无明显致Hp-T细胞凋亡的作用. AB抗血清能部分阻断Hp与胃癌细胞系的黏附,在光镜下表现为经抗AB兔血清预处理后,每细胞周围黏附的细菌数较免疫前兔血清预处理组显著减少(P<0.05).研究表明AB是安全的、具有免疫原性的Hp菌体成分,既可以刺激体液免疫,又能够提高细胞免疫,并且其抗体还可防止Hp与胃上皮细胞的黏附.
To evaluate its safety, biological activity and immunogenicity of the recombinant H.pylori (Hp) adhesin conservation region(AB) in vitro so that to determine the feasibility of AB in Hp vaccination. ELISA assay was used to measure AB-specific antibody in serum of Hp infected patients. The proliferation of T cell in response to AB was examined by MTT test. Flow cytometry was used to evaluate the increase in FasL expression on T cells under the stimulation of Hp AB,T cell apoptosis induced by AB was detected by DAP assay.The effect of anti-AB serum on Hp binding of human gastric carcinoma cell lines was determined by light microscopy. Antibodies were detected in sera samples from 55 patients by ELISA, with RUT as parallel compare. Kappa value is 0.76. The low dose AB was capable of stimulating proliferation of T cells from Hp positive subjects. The effect of AB was significantly lower in both the induction of apoptosis and FasL expression of T cells than that of ATCC26695. Anti-AB serum could partially inhibit Hp binding to gastric epithelial cells. Under light microscopy, the adhesion of Hp to MGC-803 was significantly inhibited when pretreated the bacteria with anti-AB rabbit serum, compared with negative control which pretreated with pre-immunized rabbit serum. AB was a safe, immunogenicity thallus component, which can stimulate humoral and cellular immunity.Its antibody was capable of preventing the binding of Hp to gastric epithelial cell.
白杨,张亚历,陈烨,王继德,周殿元.幽门螺杆菌黏附素保守区蛋白的免疫原性、安全性和黏附作用的体外评价[J].生物化学与生物物理进展,2003,30(3):422-426
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