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利用单克隆抗体对西西里斑点热群立克次体菌株进行鉴定

Characterization of Sicilian strains of spotted fever group rickettsiae by using monoclonal antibodies.

作者信息

Vitale G, Di Stefano R, Damiani G, Mansueto S

机构信息

Cattedra di Patologia Speciale Medica e Metodologia Clinica, Piazza delle Cliniche, Palermo, Italy.

出版信息

J Clin Microbiol. 1989 May;27(5):1081-5. doi: 10.1128/jcm.27.5.1081-1085.1989.

Abstract

Twenty-two hybridomas producing anti-Rickettsia conorii monoclonal antibodies were obtained by nine fusion experiments. The strain chosen for immunization of mice was MAVI, an R. conorii strain isolated from a Sicilian patient with Boutonneuse fever. When tested for immunoglobulin isotype by an indirect immunofluorescence (IIF) assay, 46.6% of supernatants from the 22 hybridomas were immunoglobulin M. The supernatants were tested in the IIF assay for binding to the MAVI strain and four spotted fever group rickettsia strains isolated from Sicilian ticks (two virulent and two nonpathogenic when inoculated intraperitoneally in male guinea pigs). Only five of the supernatants showed a positive IIF result on all tested strains, although they produced different titers to the various strains, possibly an indication that they recognized an antigen common to spotted fever group rickettsiae. Immunodominant epitopes for humans were determined by using patient sera to analyze inhibition of binding to the MAVI strain. Although a limited number of serum samples were screened, a high percentage of Boutonneuse fever patients produced antibodies recognizing the same epitopes as were recognized by the mouse monoclonal antibodies. A striking heterogeneity was found both in the expression of mouse-recognized epitopes on the five rickettsial strains and in the serum antibody responses of Boutonneuse fever patients to these epitopes.

摘要

通过九次融合实验获得了22株产生抗康氏立克次体单克隆抗体的杂交瘤。用于免疫小鼠的菌株是MAVI,这是一种从一名患纽扣热的西西里患者分离出的康氏立克次体菌株。通过间接免疫荧光(IIF)试验检测免疫球蛋白同种型时,22株杂交瘤中有46.6%的上清液为免疫球蛋白M。在IIF试验中检测上清液与MAVI菌株以及从西西里蜱中分离出的四种斑点热群立克次体菌株(腹腔注射雄性豚鼠时,两种有毒,两种无毒)的结合情况。尽管上清液对不同菌株产生的滴度不同,但只有五种上清液在所有测试菌株上显示IIF结果为阳性,这可能表明它们识别斑点热群立克次体共有的一种抗原。通过使用患者血清分析对与MAVI菌株结合的抑制作用来确定人类的免疫显性表位。尽管筛选的血清样本数量有限,但高比例的纽扣热患者产生的抗体识别与小鼠单克隆抗体相同表位。在五种立克次体菌株上小鼠识别的表位表达以及纽扣热患者对这些表位的血清抗体反应中均发现了显著的异质性。

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