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针对镰形艾美耳球虫并与两种禽球虫的子孢子发生交叉反应的单克隆抗体的制备与特性分析

Production and characterization of monoclonal antibodies directed against Eimeria falciformis and cross-reactive with sporozoites from two species of avian coccidia.

作者信息

Kazanji M, Rhalem A, Péry P

机构信息

Unité de Virologie et Immunologie Moléculaires, INRA, Jouy-en-Josas, France.

出版信息

Parasitol Res. 1994;80(5):396-402. doi: 10.1007/BF00932377.

Abstract

Monoclonal antibodies (mAbs) were obtained against the surface antigens of the Eimeria falciformis sporozoite by immunizing mice with whole homogenized sporozoite. The hybridomas were selected by their reactivities against oocyst extracts, then against glutaraldehyde-treated sporozoites. Three mAbs recognized both the surface of E. falciformis, E. tenella, and E. acervulina and their refractile bodies, whereas a fourth mAb recognized only one epitope on the refractile bodies. All mAbs bound to the same immunoaffinity-purified antigens in Western-blot analysis (P27 for E. falciformis and P25 for E. tenella and E. acervulina). Thus, the mAbs define at least two shared epitopes between sporozoite antigens from different eimerian species. Two of these mAbs are involved in the in vitro phagocytosis of E. falciformis sporozoites by macrophages and also in their lysis by neutrophils. Altogether, these properties showed that the four mAbs came from different activated B-cells. The P27 antigen recognized by our mAbs represents a major target of the in vitro destructive immune response.

摘要

通过用全匀浆子孢子免疫小鼠,获得了针对镰形艾美耳球虫子孢子表面抗原的单克隆抗体。通过杂交瘤对卵囊提取物的反应性进行筛选,然后对经戊二醛处理的子孢子进行反应性筛选。三种单克隆抗体识别镰形艾美耳球虫、柔嫩艾美耳球虫和堆型艾美耳球虫的表面及其折光体,而第四种单克隆抗体仅识别折光体上的一个表位。在蛋白质印迹分析中,所有单克隆抗体均与相同的免疫亲和纯化抗原结合(镰形艾美耳球虫的P27以及柔嫩艾美耳球虫和堆型艾美耳球虫的P25)。因此,这些单克隆抗体定义了来自不同艾美耳属物种的子孢子抗原之间至少两个共同表位。其中两种单克隆抗体参与巨噬细胞对镰形艾美耳球虫子孢子的体外吞噬作用以及中性粒细胞对它们的裂解作用。总之,这些特性表明这四种单克隆抗体来自不同的活化B细胞。我们的单克隆抗体识别的P27抗原是体外破坏性免疫反应的主要靶点。

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