摘要
目的:制备高效价E型沙眼衣原体(Ct)主要外膜蛋白(MOMP)168多表位特异性免疫血清并研究其免疫学特性。方法:纯化的Trx-his/Ct MOMP168融合蛋白经弗氏佐剂乳化后,小鼠背部皮下多点免疫注射,间隔2周后加强免疫,共3次,并分别于免疫前和每次免疫后2周取血,采用ELISA法检测血清融合蛋白和E型Ct全菌体特异性抗体水平及变化趋势,通过体外中和反应检测多表位特异性血清的免疫学特性。结果:小鼠用Ct MOMP168多表位融合蛋白全程免疫后产生了抗融合蛋白和E型Ct全菌体特异性抗体,ELISA法检测最高滴度达1:500。ELISA和体外中和实验检测结果显示,该抗体能特异性识别MOMP多表位蛋白和E型Ct全菌体,并能有效抑制E型Ct感染Hela229细胞。结论:沙眼衣原体MOMP多表位融合蛋白免疫血清可特异性结合并中和E型Ct。
Objective：To prepare high-titer immune serum specifically against multi-epitope fusion protein of Chlamydia trachomatis（Ct） major outer membrane protein（MOMP168） and analyze its immunological characteristics.Methods： BALB/c mice were subcutaneously immunized with the purified Trx-his/Ct MOMP168 protein formulated with Freund adjuvant.Three vaccinations were given at 2-week intervals.Sera were taken prior to the first immunization and 2 weeks after each immunization.The specific antibodies to the fusion protein and the whole cell of Ct were assayed with ELISA.The immunological feature of immunized sera was tested by in vitro neutralization assays.Results： All immunized mice produced high-titer specific antibody to the fusion protein and the whole cell of Ct serotype E after full immunizations.The highest antibody titer was arrived at 1：500.As measured with ELISA and in vitro neutral-ization assays,specific antibodies induced not only perfectly recognized the Trx-his/Ct MOMP168 fusion protein and the whole cell of Ct serotype E,but also displayed a significant inhibitory effect on the invasion of Ct serotype E to Hela229 cell.Conclusion： The immune sera against the Ct MOMP multi-epitope fusion protein can specifically bind Ct of serovar E and show neutralization activity.
出处
《温州医学院学报》
CAS
2012年第1期6-9,共4页
Journal of Wenzhou Medical College
基金
国家自然科学基金资助项目(30972669)
关键词
沙眼衣原体
主要外膜蛋白
表位
中和作用
Chlamydia trachomatis
major outer membrane protein（MOMP）
epitope
neutral-ization
