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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 and characterize multi-epitope specific immunity serum of high-titer Chlamydia trachomatis (Ct) 168 major outer membrane protein (MOMP). Methods: The purified Trx-his / Ct MOMP168 fusion protein was emulsified with Freund’s adjuvant. Mice were immunized subcutaneously with multiple immunization subcutaneously twice a week for 3 times before immunization and after each immunization The blood samples were collected for 2 weeks. The levels of serum fusion proteins and whole-body-specific antibodies of E-type Ct were determined by ELISA. The immunological characteristics of multi-epitope-specific serum were detected by in vitro neutralization. Results: Mice immunized with Ct MOMP168 multi-epitope fusion protein produced full-fledged anti-fusion protein and whole-cell antibody against type E Ct. The highest titer was 1: 500 by ELISA. ELISA and in vitro neutralization test results show that the antibody can specifically recognize MOMP polytope protein and E-type Ct whole cell, and can effectively inhibit E-Ct infection Hela229 cells. CONCLUSION: Chlamydia trachomatis MOMP multi-epitope fusion protein can specifically bind to and neutralize type E Ct.