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利用单克隆抗体对胎儿三毛滴虫抗原进行鉴定

Characterization of Tritrichomonas foetus antigens by use of monoclonal antibodies.

作者信息

Hodgson J L, Jones D W, Widders P R, Corbeil L B

机构信息

Department of Veterinary Microbiology and Pathology, Washington State University, Pullman 99164-7040.

出版信息

Infect Immun. 1990 Sep;58(9):3078-83. doi: 10.1128/iai.58.9.3078-3083.1990.

Abstract

The specificity for and function of monoclonal antibodies against Tritrichomonas foetus were characterized. Four monoclonal antibodies generated by immunization of mice with live T. foetus were selected on the basis of enzyme-linked immunosorbent assay reactions. The approximate molecular masses of the predominant proteins were determined by Western blotting (immunoblotting). Monoclonal antibody TF3.8 recognized a predominant band at approximately 155 kilodaltons, whereas TF3.2 reacted with several bands. Monoclonal antibodies TF1.17 and TF1.15 recognized broad bands between 45 and 75 kilodaltons. The first two antibodies (TF3.8 and TF3.2) did not react with the surface of T. foetus, as determined by live-cell immunofluorescence, agglutination, and immobilization, whereas two other monoclonal antibodies (TF1.17 and TF1.15) did react with surface epitopes, as determined by these criteria. The latter two monoclonal antibodies also mediated complement-dependent killing of T. foetus and prevented of adherence of organisms to bovine vaginal epithelial cells. One antibody, TF1.15, also killed in the absence of complement. Since these functions are in vitro correlates of protection, the antigens recognized by these monoclonal antibodies may induce protective immunity.

摘要

对针对胎儿三毛滴虫的单克隆抗体的特异性和功能进行了表征。基于酶联免疫吸附测定反应,选择了用活的胎儿三毛滴虫免疫小鼠产生的四种单克隆抗体。通过蛋白质印迹法(免疫印迹)测定主要蛋白质的近似分子量。单克隆抗体TF3.8识别出一条约155千道尔顿的主要条带,而TF3.2与几条条带发生反应。单克隆抗体TF1.17和TF1.15识别出45至75千道尔顿之间的宽带。通过活细胞免疫荧光、凝集和固定化测定,前两种抗体(TF3.8和TF3.2)不与胎儿三毛滴虫的表面发生反应,而另外两种单克隆抗体(TF1.17和TF1.15)根据这些标准确实与表面表位发生反应。后两种单克隆抗体还介导了补体依赖性的胎儿三毛滴虫杀伤作用,并阻止了生物体与牛阴道上皮细胞的粘附。一种抗体TF1.15在没有补体的情况下也具有杀伤作用。由于这些功能是体外保护作用的相关指标,这些单克隆抗体识别的抗原可能诱导保护性免疫。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4415/313614/a9f59ff19f30/iai00057-0366-a.jpg

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