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用荧光素偶联凝集素检测卡氏肺孢子虫表面碳水化合物

Detection of surface carbohydrates on Pneumocystis carinii by fluorescein-conjugated lectins.

作者信息

Yoshikawa H, Tegoshi T, Yoshida Y

机构信息

Department of Medical Zoology, Kyoto Prefectural University of Medicine, Japan.

出版信息

Parasitol Res. 1987;74(1):43-9. doi: 10.1007/BF00534931.

Abstract

Lectins react with a wide range of different carbohydrates (Table 1). Even so-called monospecific anti-H(O) lectins from Lotus tetragonolobus, Ulex europaeus, and Anguilla anguilla react not only with the anti-H determinant but also with several fucosylated carbohydrates. Consequently, the type of lectin receptor existing on the surface of Pneumocystis carinii should be determined, because only a carbohydrate analysis can demonstrate the kind of carbohydrates which exist on the cell surface of this parasite. For the purpose of this study we used fluorescent isothiocyanata (FITC)-conjugated lectins. Concanavalin A (Con A) and Maclura pomifera (MPA) agglutinin reacted to P. carinii at low concentrations, and the fluorescence intensity was gradually increased with the concentration of the lectins. With lectins from Bauhinia purpurea (BPA), Dolichos biflorus (DBA), Glycine max (SBA), Griffonia simplicifolia (GS-I, GS-II), and Triticum vulgaris (WGA), fluorescence was emitted at high concentrations, while Arachis hypogaea (PNA) and Ulex europaeus (UEA-I) agglutinins did not show fluorescence. The results suggest that P. carinii has abundant Con A- and MPA-specific carbohydrates on the surface.

摘要

凝集素能与多种不同的碳水化合物发生反应(表1)。即便如此,来自四角豆、荆豆和欧洲鳗鲡的所谓单特异性抗H(O)凝集素不仅能与抗H决定簇发生反应,还能与几种岩藻糖基化碳水化合物发生反应。因此,应确定卡氏肺孢子虫表面存在的凝集素受体类型,因为只有碳水化合物分析才能证明该寄生虫细胞表面存在的碳水化合物种类。为了本研究的目的,我们使用了异硫氰酸荧光素(FITC)偶联的凝集素。刀豆球蛋白A(Con A)和桑科柘属植物(MPA)凝集素在低浓度下就能与卡氏肺孢子虫发生反应,且荧光强度会随着凝集素浓度的增加而逐渐增强。对于来自紫羊蹄甲(BPA)、双花扁豆(DBA)、大豆(SBA)、非洲血根草(GS-I、GS-II)和普通小麦(WGA)的凝集素,在高浓度时会发出荧光,而花生(PNA)和荆豆(UEA-I)凝集素则不显示荧光。结果表明,卡氏肺孢子虫表面有丰富的Con A和MPA特异性碳水化合物。

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