幽门螺杆菌表位疫苗的设计、构建、表达及其免疫原性研究  

Design, construction, expression, and immunogenicity of Helicobacter pylori epitope vaccine

作　　者：毛旭虎[1] 石云[1] 吴超[1] 张卫军[1] 邹全明[1] 

机构地区：[1]第三军医大学医学检验系临床微生物学及免疫学教研室重庆市生物制药工程技术研究中心,重庆400038

出　　处：《免疫学杂志》2006年第4期381-383,386,共4页Immunological Journal

基　　金：国家自然科学基金资助项目(30400406)

摘　　要：目的设计幽门螺杆菌(Hp)的表位疫苗并对其免疫原性进行研究。方法将Hp的尿素酶B亚单位的三个Th表位及一个B细胞表位串联,表位之间用两个赖氨酸(KK)间隔,合成全基因,克隆到pET22b载体上,在大肠杆菌BL21(DE3)中表达;表达的重组蛋白经鉴定纯化后皮下免疫Balbc小鼠,检测细胞免疫应答及体液免疫应答。结果克隆表达的重组表位疫苗蛋白纯化后纯度达到95%,经N端测序证实为设计的表位疫苗蛋白,Th表位多肽(U546-561、U229-244、U237-251)、表位疫苗蛋白及rUreB均能够刺激表位疫苗致敏的小鼠淋巴细胞增殖(SI>2),并且此疫苗能够刺激机体产生特异性抗体。结论本研究中表位疫苗的设计方法能够使疫苗中包含的四个表位均发挥功能,并且具有较强的免疫原性,为研究Hp预防性和治疗性疫苗提供实验基础。Objective To design the Helicobacter pylori epitope vaccine and study its immunogenicity in Balb/c mice. Methods The epigene was designed and synthesized, containing three Th cell epitopes and one B cell epitope that come from urease B subunit. The epigene was cloned into expression vector pET-22b（ ＋ ） and expressed in Escherichia coliBL21 （DE3）. Balb/c mice were immunized subcutaneously by the purified protein and then the cellular and humoral immune response was detected. Results Recombinant protein was successfully expressed in Escherichia coli BL21 and could elicit strong cellular and humoral immune response. Conclusion The method of designing the epitope vaccine is proved to be successful and the expressed protein way be a potential candidate of tberapeutic and preventive vaccine of Helicobacter pylori.

关 键 词：幽门螺杆菌 尿素酶 B亚单位 表位 疫苗 

分 类 号：R392[医药卫生—免疫学]