幽门螺杆菌黏附素HpaA蛋白在大肠杆菌中表达及其免疫保护作用

目的表达和纯化幽门螺杆菌(Helicobacter pylori,H.pylori)黏附素HpaA蛋白,研究其免疫活性和对小鼠的免疫保护作用。方法诱导TB1(pMAL-c2X-hpaA)表达H.pylori黏附素HpaA蛋白,Amyloss树脂预装柱进行分离、纯化,Western blot鉴定其免疫学活性;纯化蛋白免疫小鼠后,H.pylori国际标准菌株NCTC11637攻击感染,观察H.pylori定植情况及免疫保护效果。结果获得了高纯度且免疫活性良好的目的蛋白;HpaA蛋白免疫小鼠的保护率为53.33%(8/15),与对照组统计学差异显著(P=0.002)。结论纯化HpaA蛋白对小鼠有免疫保护作用,可作为H.pylori基因工程疫苗的候选组分。

Expression of HpaA protein from Helicobacter pylori in E.coli and its Immunoprotection

The objective of this study was to express and purify the HpaA protein of Helicobacter pylori（H.pylori）,and to investigate its immunogenicity and the immunoprotection of mice against H.pylori infection.TB1（pMAL-c2X-hpaA）were induced by IPTG to express HpaA protein of H.pylori.The soluble recombinant HpaA protein was purified by amyloss pre-packed column and the purity was detected by Western blot.After immunizing with HpaA protein,mice were challenged by H.pylori strains NCTC11637 and the gastric H.pylori infection was observed.Results displayed that the purified HpaA protein was confirmed to have high immunogenicity.The protective rate of HpaA protein immunized mice was 53.33%（8/15）and significantly higher than that of control group（P =0.002）.The highly purified HpaA protein showed good immunogenicity and immunoprotection in this study and could be used for further development of H.pylori vaccine.