Production and characterization of species-specific monoclonal antibodies against Leishmania donovani for immunodiagnosis.
Production and characterization of species-specific monoclonal antibodies against Leishmania donovani for immunodiagnosis.
复制标题
用于免疫诊断的针对杜氏利什曼原虫的物种特异性单克隆抗体的生产和表征。
作者：
C. Jaffe;E. Bennett;G. Grimaldi;D. Mcmahon
制备了16种针对多诺瓦利什曼原虫膜的单克隆抗体。这些抗体只与多诺瓦氏乳杆菌上的决定因子反应。未发现与其他利什曼原虫或克氏锥虫膜发生交叉反应。通过使用整个promastigote匀浆作为抗原，对选定抗体的结合特异性进行了广泛的分析。单克隆抗体D-1、D-2、D-3和D-4从84个新旧世界利什曼原虫种群的交叉面板中正确识别了所有44个多诺瓦尼利什曼原虫种群。抗体D-1和D-2也可用于免疫荧光分类。未观察到与所检查的任何其他利什曼原虫种发生交叉反应。基于Western blot和/或放射免疫沉淀分析，鉴定出与L. donovani特异性抗原决定因子相关的五组不同的分子。这些分子的毫瓦从18到84千道尔顿不等。被D-2、D-10和D-13抗体识别的抗原分子也被内脏利什曼病（黑热病）患者血清中存在的抗体识别。在Western blot分析中，从旧世界和新世界的病例中获得的黑热病血清与这些单克隆抗体特异性竞争，以获得适当的抗原决定因子。这些单克隆抗体和/或纯化的蛋白抗原可用于发展血清学检测，用于临床诊断由多诺瓦氏乳杆菌引起的内脏利什曼病和利什曼病的流行病学研究。
Sixteen species-specific monoclonal antibodies were produced against membranes of Leishmania donovani. These antibodies only reacted with determinants present on L. donovani. No cross-reactions were found with any other species of Leishmania or with membranes of Trypanosoma cruzi. An extensive analysis of the binding specificities of selected antibodies was carried out by using whole promastigote homogenates as antigen. Monoclonal antibodies D-1, D-2, D-3, and D-4 correctly identified all 44 L. donovani stocks from a cross-panel of 84 New and Old World Leishmania stocks. Antibodies D-1 and D-2 were also useful for species classification by immunofluorescence. No cross-reactions were observed with any other Leishmania species examined. Based on either Western blot and/or radioimmunoprecipitation analyses, five distinct groups of molecules associated with L. donovani-specific antigenic determinants were identified. These molecules range in m.w. from 18 to 84 kilodaltons. The antigenic molecules recognized by antibodies D-2, D-10, and D-13 are also recognized by antibodies present in sera from patients with visceral leishmaniasis (kala-azar). Kala-azar sera obtained from cases in both the Old and New World specifically compete with these monoclonal antibodies for the appropriate antigenic determinants in Western blot analysis. These monoclonal antibodies and/or the purified protein antigens may be useful in the development of a serologic assay for the clinical diagnosis of visceral leishmaniasis caused by L. donovani and in epidemiologic studies of leishmaniasis.