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单克隆抗体可识别微小隐孢子虫子孢子和配子体中致密颗粒的一个亚群。

Monoclonal antibodies identify a subset of dense granules in Cryptosporidium parvum zoites and gamonts.

作者信息

Bonnin A, Gut J, Dubremetz J F, Nelson R G, Camerlynck P

机构信息

Laboratoire de Parasitologie-Mycologie, Hôpital du Bocage, Dijon, France.

出版信息

J Eukaryot Microbiol. 1995 Jul-Aug;42(4):395-401. doi: 10.1111/j.1550-7408.1995.tb01601.x.

Abstract

Two monoclonal antibodies raised against purified oocysts and excysted sporozoites of Cryptosporidium parvum identified antigens located in the anterior half of sporozoites by indirect immunofluorescence microscopic assay. The monoclonal antibodies also reacted with Triton-X-100-insoluble antigens of asexual and sexual stage parasites developing in epithelial cells in vitro and identified a 110 kilodalton antigen on immunoblots of sodium dodecyl sulfate-extracted oocysts. Immunoblotting reactivity was abolished by prior treatment of blotted antigen with periodic acid suggesting that the monoclonal antibodies recognize a carbohydrate or carbohydrate-dependent epitope(s). By immunoelectron microscopy, the antibodies reacted with a family of small, electron-dense granules located predominantly in the central region of merozoites and also with a population of cytoplasmic inclusions in macrogamonts. In addition, the monoclonal antibodies prominently labeled the parasitophorous vacuole membrane of all intracellular stages examined suggesting that the corresponding antigen(s) may be exocytosed from the granules to become associated with Triton X-100-insoluble components of the vacuolar membrane or cytoskeleton.

摘要

两种针对纯化的微小隐孢子虫卵囊和脱囊子孢子产生的单克隆抗体,通过间接免疫荧光显微镜检测法,鉴定出位于子孢子前半部分的抗原。这些单克隆抗体还与体外在上皮细胞中发育的无性和有性阶段寄生虫的Triton-X-100不溶性抗原发生反应,并在十二烷基硫酸钠提取的卵囊的免疫印迹上鉴定出一种110千道尔顿的抗原。通过先用高碘酸处理印迹抗原,免疫印迹反应性消失,这表明单克隆抗体识别碳水化合物或碳水化合物依赖性表位。通过免疫电子显微镜观察,这些抗体与主要位于裂殖子中心区域的一族小的、电子致密颗粒发生反应,也与大配子体中的一群胞质内含物发生反应。此外,单克隆抗体显著标记了所检测的所有细胞内阶段的寄生泡膜,这表明相应的抗原可能从颗粒中胞吐出来,与泡膜或细胞骨架的Triton X-100不溶性成分结合。

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