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单克隆抗体识别的保护性蛋白免疫原在多杀性巴氏杆菌兔分离株外膜上的分布

Distribution of a monoclonal antibody-recognized protective protein immunogen on the outer membranes of Pasteurella multocida rabbit isolates.

作者信息

Lu Y S, Gerrity L W, Afendis S J, Watkins L, Pakes S P

机构信息

Department of Pathology, University of Texas Southwestern Medical Center, Dallas 75235.

出版信息

J Clin Microbiol. 1988 Jul;26(7):1326-30. doi: 10.1128/jcm.26.7.1326-1330.1988.

Abstract

The distribution of a monoclonal antibody (MAb)-recognized protective protein immunogen on the outer membrane of 153 Pasteurella multocida rabbit isolates was determined by dot blot (DB) analysis. MAb 1608 reacted with 36 (24%) of the 153 clinical isolates. The DB-positive clinical isolates expressed capsular antigens A, D, and nontypable and somatic antigens 2, 3, 10, 12, 15, and nontypable. Western blot (immunoblot) analysis with adsorbed and eluted MAb 1608 confirmed that the antigenic determinant identified was located on the cell surface. With MAb 1608 as a probe for antibody-accessible radioimmunoassay, 31 of 36 DB-positive P. multocida rabbit isolates were shown to have surface-exposed and antibody-accessible antigenic determinants, while 44 of 44 DB-negative isolates were negative by antibody-accessible radioimmunoassay. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed DB-negative P. multocida isolates both with (6 of 13, 46%) and without (7 of 13, 54%) the 37.5-kilodalton protein. This study establishes that the protective antigenic determinant of the 37.5-kilodalton outer membrane protein is present in 24% of rabbit clinical isolates tested and is detectable in P. multocida strains distributed among the major somatic types (3, 10, 12, and 15) and the capsular types (A and D) commonly isolated from rabbits in North America.

摘要

通过斑点印迹(DB)分析确定了单克隆抗体(MAb)识别的保护性蛋白免疫原在153株多杀性巴氏杆菌兔分离株外膜上的分布。MAb 1608与153株临床分离株中的36株(24%)发生反应。DB阳性临床分离株表达荚膜抗原A、D和不可分型以及菌体抗原2、3、10、12、15和不可分型。用吸附和洗脱的MAb 1608进行蛋白质印迹(免疫印迹)分析证实,所鉴定的抗原决定簇位于细胞表面。以MAb 1608作为抗体可及性放射免疫测定的探针,36株DB阳性多杀性巴氏杆菌兔分离株中有31株显示有表面暴露且抗体可及的抗原决定簇,而44株DB阴性分离株在抗体可及性放射免疫测定中均为阴性。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示,DB阴性的多杀性巴氏杆菌分离株中既有(13株中的6株,46%)也有(13株中的7株,54%)37.5千道尔顿蛋白。本研究证实,37.5千道尔顿外膜蛋白的保护性抗原决定簇存在于24%的受试兔临床分离株中,并且在北美常见的从兔中分离出的多杀性巴氏杆菌菌株中,分布于主要菌体类型(3、10、12和I5)和荚膜类型(A和D)的菌株中均可检测到。

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