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柔嫩艾美耳球虫:一种单克隆抗体在确定折光体细胞器的细胞内命运及对体外发育影响方面的应用

Eimeria tenella: use of a monoclonal antibody in determining the intracellular fate of the refractile body organelles and the effect on in vitro development.

作者信息

Danforth H D, Augustine P C

机构信息

Protozoan Diseases Laboratory, Livestock and Poultry Sciences, Beltsville, Maryland 20705.

出版信息

Exp Parasitol. 1989 Jan;68(1):1-7. doi: 10.1016/0014-4894(89)90002-7.

DOI:10.1016/0014-4894(89)90002-7
PMID:2645159
Abstract

A monoclonal antibody, which recognizes the refractile body of Eimeria sporozoites, was used to study the developmental fate of this organelle during asexual development of E. tenella and to determine the effect of this monoclonal antibody on in vitro development of the parasite. Through use of immunofluorescent antibody and gold-labeling techniques at the light and electron microscopy level, the refractile body at 48 to 96 hr postinoculation was found to separate into 6 to 10 small globules, then diffuse throughout the schizont cytoplasm, and eventually reconcentrate as a small dot of material in each of the mature first-generation merozoites. The schizont did not develop to maturity if diffusion of the refractile body did not occur. The refractile body material was quickly lost as the merozoite left the schizont and invaded new cells and was not detected in any later developmental stages. The in vitro development of first- and second-generation schizonts of E. tenella was greatly inhibited (up to 100%) with exposure to the monoclonal antibody. There was an increase in the number of schizonts with nondispersed refractile body in the monoclonal antibody-treated cells when compared to the untreated controls, and the few mature schizonts seen had up to a 50-fold decrease in the number of merozoites. Immunofluorescent antibody labeling of the refractile body of intracellular sporozoites and schizonts treated in vitro with the monoclonal antibody for 24-96 hr postinoculation indicated that the antibody had crossed the host cell and parasite plasma membrane during incubation.

摘要

一种识别艾美耳球虫子孢子折光体的单克隆抗体,被用于研究该细胞器在柔嫩艾美耳球虫无性发育过程中的发育命运,并确定这种单克隆抗体对该寄生虫体外发育的影响。通过在光学和电子显微镜水平上使用免疫荧光抗体和金标技术,发现在接种后48至96小时的折光体分离成6至10个小球,然后扩散到裂殖体细胞质中,并最终在每个成熟的第一代裂殖子中重新浓缩为一小点物质。如果折光体不发生扩散,裂殖体就不会发育成熟。随着裂殖子离开裂殖体并侵入新细胞,折光体物质迅速丢失,在任何后期发育阶段都未检测到。暴露于单克隆抗体后,柔嫩艾美耳球虫第一代和第二代裂殖体的体外发育受到极大抑制(高达100%)。与未处理的对照相比,在单克隆抗体处理的细胞中,具有未分散折光体的裂殖体数量增加,并且所见到的少数成熟裂殖体中的裂殖子数量减少了多达50倍。对接种后用单克隆抗体体外处理24 - 96小时的细胞内子孢子和裂殖体的折光体进行免疫荧光抗体标记表明,在孵育过程中抗体已穿过宿主细胞和寄生虫的质膜。

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