幽门螺杆菌UreB蛋白对小鼠的免疫调节作用

Effects of Helicobacter pylori UreB protein on immunity regulation of mice

目的鉴定幽门螺杆菌(Helicobacter pylori,H.pylori)UreB蛋白对小鼠的免疫调节作用,为H.pylori致病机制和免疫防治研究奠定基础。方法利用前期纯化制备的由大肠杆菌(Escherichia Coli,E.coli)TB1/pMAL-c2x-UreB表达的H.pyloriUreB重组蛋白(rUreB),对实验组小鼠进行背部皮下注射,对照组采用蛋白纯化过柱缓冲液(Column buffer)替代重组蛋白注射液。在注射后1周和2周采集小鼠血液样品,进行血细胞常规计数和淋巴细胞流式分析。对比分析实验组与对照组小鼠血液免疫细胞构成比的差异。结果全血细胞计数显示,实验组红细胞(RBC)和白细胞(WBC)计数与对照组比较,差异无统计学意义(P>0.05);在rUreB免疫后1周,小鼠外周血CD4^-CD3^+T细胞和CD3^+T细胞在总淋巴细胞中所占构成比均低于对照组,差异有统计学意义(P<0.05);2周后,组间差异无统计学意义(P>0.05);CD4^+CD3^+T淋巴细胞与总淋巴细胞比值、CD4^+CD3^+与CD4^-CD3^+T淋巴细胞比值在免疫组与对照组间的差异无统计学意义(P>0.05)。结论H.pyloriUreB经皮下注射可改变淋巴细胞亚群的构成比,且该效应具有明显时效性特征。该研究发现,对H.pylori感染免疫和致病机制的研究具有参考价值。

Objective To identify effect of Helicobacter pylori ( H.pylori ) UreB protein on immune regulation of mice and aim to lay basis for approaches on pathogenesis and immune prevention of H.pylori infection. Methods Using recombinant UreB expressed in Escherichia Coli ( E.coli ) TB1/pMAL-c2x-UreB and purified previously,mice of experimental group were immunized by subcutaneous injection into the back.For those of the controls,column buffer for the protein purification was used instead of the protein solution.One and two weeks after injection,blood of the mice was sampled and blood cell counting and flow cytometry were carried out.The difference in blood immunocyte component ratio between two groups was analyzed. Results It was determined that there was no statistically significant difference existed between experimental and control group of blood cell counts. One week of blood cell counts after the immunization with UreB,the component ratios of CD4^- CD3^+ T cells and that of CD3^+ T cells in total lymphocytes in experimental group were significantly lower than those in control group ( P <0.05).Nevertheless,the difference became statistically insignificant two weeks after infection.The difference in the ratios of CD3^+ CD4^+ T/total lymphocytes,CD4^+ CD3^+ T/CD4^- CD3^+ T was insignificant between two groups ( P >0.05). Conclusion The component ratios of certain lymphocyte subgroups in total lymphocytes can be changed by subcutaneous injection of H.pylori UreB,and the effects have apparent characters of timeliness.The findings are of reference value for researches on immunization and pathogenesis mechanisms of H.pylori infection.