沙眼衣原体蛋白水解酶CT841在感染细胞中的定位及抗原性研究  

Localization of Chlamydia protease CT841 in infected cells and the antigenicity analysis of the protein

作　　者：陆春雪[1] 曾浩[2] 吴移谋[1] 彭波[1] 胡四海[1] 蔡恒玲[1] 李忠玉[1] 钟光明[3] 

机构地区：[1]南华大学微生物与免疫学教研室,衡阳421001 [2]第三军医大学临床微生物学及免疫学教研室,重庆400038 [3]美国德克萨斯大学圣安东尼奥健康科学中心微生物学与免疫学系,德州78229

出　　处：《免疫学杂志》2012年第3期203-207,共5页Immunological Journal

基　　金：国家自然科学基金(81102230);湖南省高校科技创新团队(湘教通[2010]53号)

摘　　要：目的分析沙眼衣原体蛋白水解酶CT841在感染细胞中的定位并探讨其抗原性。方法利用PCR技术获得衣原体CT841基因,将基因序列克隆到载体pGEX6p,转化大肠杆菌XL1-blue,IPTG诱导表达融合蛋白GST-CT841。融合蛋白经纯化后免疫小鼠制备特异性抗体,间接免疫荧光法分析CT841在感染细胞中的分布特征;ELISA法分析CT841的抗原性。结果CT841原核表达重组体成功构建;CT841在感染细胞的表达模式与主要外膜蛋白MOMP相似,而与衣原体蛋白酶样活性因子CPAF及包涵体膜蛋白IncA的分布模式不同;CT841与衣原体感染患者、猴、鼠血清均发生强烈的免疫反应。结论沙眼衣原体蛋白酶CT841是定位于衣原体菌体上的免疫优势抗原。This study aims to localize protease CT841 in Chlamydia trachomatis-infected cells and investigate the antigenicity of the protein.Firstly,Chlamydia trachomatis CT841 gene was amplified by PCR and insert into pGEX-6p vector,and then the constructed vector was transformed into E.coli XL1Blue and induced by IPTG.After purified with Glutathione Sepharose 4B beads,the CT841 fusion protein was used to immunize mice to produce polyclone antibody,which were subsequently used to localize the endogenous CT841 protein by indirect immunofluorescence assay（IFA）.At same time,ELISA was used to determine the reactivity of CT841 with sera samples collected from chlamydia infected patients,nonhuman primates or mice.These methods illustrated that the cellular distribution pattern of the protease CT841 is similar to that of the major outer membrane protein（MOMP） and was highly recognized by chlamydia infected antisera.All results indicated that protease CT841 is localized on the bacterial organism in C.trachomatis-infected cells and it is an immunodominant antigen of chlamydia.

关 键 词：沙眼衣原体 CT841 细胞定位 抗原性 

分 类 号：R392.3[医药卫生—免疫学]