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B群Nm 3407菌株外膜蛋白复合物 OMPC 的提取及免疫原性 |
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孙银燕 胡绪敬
【摘要】目的 B群脑膜炎奈瑟氏菌Nm荚膜多糖作为菌苗候选成分效果不理想，有些非荚膜的抗原成分存在血清型和亚型的限制，为了能提取一种免疫原性强、并具有广泛的抗原交叉反应性的B群Nm外膜蛋白复合物（outer membrane protein complex, OMPC）以研制更有效的B群Nm菌苗提供依据。方法 比较了从病人分离的B群Nm 3407菌株提取OMPC的3种方法，用ELISA、杀菌力试验、小鼠免疫试验、Western blotting测定OMPC的抗原滴度和免疫原性。结果 在提取OMPC的3种方法中，选用70％饱和硫酸铵初步沉淀细菌培养物的上清液，再经Sephacryl S-300HR纯化的方法,获得了3个洗脱峰。其中第一个洗脱峰中OMPC产量最高，溶解性最好。经SDS-PAGE及Western blotting测定，其主要含有分子量为(42，39，30，26和19)×103的蛋白和少量约(92和72)×103的蛋白。用ELISA测定此OMPC与兔抗菌株3407和542852血清反应的滴度分别为1∶32000和1∶64000；与小鼠抗此OMPC血清反应的滴度为1∶25600。另外，上述抗OMPC血清与菌株3407和542852反应的滴度均为1∶25600，与所试的几株不同型/亚型的B群Nm菌株反应滴度为1∶640～1∶3200。此OMPC免疫血清对菌株3407和542852的杀菌滴度均为1∶256。其它两种方法所提纯的OMPC其抗原性和免疫原性均较差。结论 上述B群Nm OMPC溶解性好、免疫原性强，而且具有广泛的抗原交叉反应性，以它作为B群Nm菌苗的候选成分或者作为偶联多糖抗原的载体是有希望的。
【主题词】 奈瑟氏球菌，脑膜炎 细菌外膜蛋白质类 免疫性 抗原抗体复合物
Extraction and immunogenicity of outer membrane protein complex from the strain 3407 of Nm serogroup B Sun Yinyan, Hu Xujing. Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Beijing 102206
【 Abstract 】 Objectives The capsular polysaccharide of Neisseria meningitidis serogroup B is not an effective components for a vaccine candidate because there are some limits of serotype and subserotype in some noncapsular antigens, it will be significant to prepare an effective group B vaccine if a high immunogenic and broadly cross reactive outer membrane protein complex (OMPC) can be extracted from Nm serogroup B. Methods Three methods for extracting OMPC from the strains 3407 of Nm serogroup B isolated from a clinical case were compared. ELISA, Bactericidal test, immunologic experiments with mice and Western-blotting were used for testing the antigenic titers and immunogenicity of OMPC. Results Among the three methods for extracting OMPC, the method that the cultural supernatant was primarily precipitated with 70% saturated ammonium sulfate and further purified on Sephacryl S-300 HR column was chosen, and three elution peaks[1] [2] [3] 下一页
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