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目的观察不同种株利什曼原虫抗原的异质性。方法培养杜氏利什曼原虫、热带利什曼原虫及婴儿利什曼原虫前鞭毛体及体外转化无鞭毛体,提取总蛋白,经SDS-PAGE电泳后,分别以抗杜氏利什曼原虫及热带利什曼原虫前鞭毛体血清进行Western blot,分析抗原的多样性。结果经SDS-PAGE电泳和考马斯亮蓝染色,杜氏利什曼原虫、热带利什曼原虫及婴儿利什曼原虫前鞭毛体及体外转化无鞭毛体总蛋白呈现出分子质量单位为12～150ku的相似蛋白带型;Western blot显示,抗杜氏利什曼原虫前鞭毛体多克隆血清识别的抗原组分与抗热带利什曼原虫前鞭毛体多克隆血清识别的抗原相似;3种利什曼原虫前鞭毛体和无鞭毛体存在48、65及35ku共同抗原,杜氏利什原虫和婴儿利什曼原虫的无鞭毛体存在38ku期特异抗原,热带利什曼原虫的前鞭毛体和无鞭毛体存在45、24ku种特异抗原。结论杜氏利什曼原虫、热带利什曼原虫及婴儿利什曼原虫间存在抗原多样性,为认识利什曼病感染免疫提供了新的视角。
Objective To observe the heterogeneity of different strains of Leishmania antigens. Methods Leishmania donovani, Leishmania tropicalis and Leishmania infantum promastigotes were cultured and transformed into amastigotes in vitro. Total protein was extracted. After SDS-PAGE electrophoresis, Leishmania promastigotes serum was subjected to Western blot to analyze antigenic diversity. Results The results of SDS-PAGE and Coomassie brilliant blue staining showed that the total protein of Leishmania donovani, Leishmania tropicalis and Leishmania infantum and the transformed lectin in vitro showed a molecular weight of 12-150 ku Similar protein bands; Western blot showed that anti-Leishmania promastigote polyclonal serum antigenic components identified with anti-Leishmania promastigotes polyclonal serum antigens identified similar; three species of Leishmania Precursor and amastigotes exist 48,65 and 35ku common antigens, there are 38ku phase specific antigen in the amastigotes of Leishmania donovani and Leishmania infantum, promastigotes and amastigotes of Leishmania tropic exist 45, 24ku kinds of specific antigen. Conclusion There are antigenic diversity between Leishmania donovani, Leishmania leishmanii and Leishmania infantum and provide a new perspective for understanding the immunity of leishmaniasis infection.