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针对立氏立克次体的单克隆抗体的制备与特性鉴定

Production and characterization of monoclonal antibodies to Rickettsia rickettsii.

作者信息

Lange J V, Walker D H

出版信息

Infect Immun. 1984 Nov;46(2):289-94. doi: 10.1128/iai.46.2.289-294.1984.

Abstract

Five mouse ascitic fluids (MAFs) containing monoclonal antibody to Rickettsia rickettsii were produced from three original fusions by murine hybridoma technology. The five MAFs were fractionated and purified; each contained monoclonal antibody of the immunoglobulin G2a subclass. Each monoclonal antibody-containing MAF was titrated by indirect immunofluorescence against three R. rickettsii isolates from humans and four other spotted fever group rickettsiae. Each MAF was also titrated in the complement fixation, latex agglutination, microagglutination, and indirect hemagglutination tests. Two of the MAFs were examined for their ability to prevent fever and rickettsemia in susceptible guinea pigs after a 1:100 dilution of each was mixed with viable R. rickettsii, and all five MAFs were titrated in the mouse toxicity phenomenon assay. All MAFs had high indirect immunofluorescence titers to the three strains of R. rickettsii (1:200,000 to 1:800,000), reduced indirect immunofluorescence titers to R. montana, and were nonreactive with R. akari, R. sibirica, and R. conorii. Each MAF was able to fix complement in the presence of spotted fever group antigen reagent and agglutinate a suspension of purified R. rickettsii, and each was negative in both the latex agglutination and the indirect hemagglutination tests. The two MAFs which were tested proved to be capable of preventing rickettsemia and death in guinea pigs, and each MAF was able to prevent death in mice at dilutions ranging from 1:40 to 1:80.

摘要

利用鼠杂交瘤技术从三次原始融合中制备了五种含有抗立氏立克次体单克隆抗体的小鼠腹水(MAF)。对这五种MAF进行了分级分离和纯化;每种都含有免疫球蛋白G2a亚类的单克隆抗体。通过间接免疫荧光法对每种含有单克隆抗体的MAF针对三株来自人类的立氏立克次体分离株和其他四株斑点热群立克次体进行了滴定。每种MAF还在补体结合、乳胶凝集、微量凝集和间接血凝试验中进行了滴定。将其中两种MAF在与活的立氏立克次体混合进行1:100稀释后,检测其预防易感豚鼠发热和立克次体血症的能力,并且对所有五种MAF进行了小鼠毒性现象测定。所有MAF对三株立氏立克次体菌株均具有高间接免疫荧光滴度(1:200,000至1:800,000),对蒙大拿立克次体的间接免疫荧光滴度降低,并且与小蛛立克次体、西伯利亚立克次体和康氏立克次体无反应。每种MAF在斑点热群抗原试剂存在下能够固定补体并凝集纯化的立氏立克次体悬液,并且在乳胶凝集和间接血凝试验中均为阴性。经检测的两种MAF被证明能够预防豚鼠的立克次体血症和死亡,并且每种MAF在1:40至1:80的稀释度下能够预防小鼠死亡。

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