Doyle P S, Engel J C, Pimenta P F, da Silva P P, Dwyer D M
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Exp Parasitol. 1991 Oct;73(3):326-34. doi: 10.1016/0014-4894(91)90104-5.
L. donovani promastigotes were subjected to heat treatment yielding an axenic amastigote stage which was long-term cultured at 37 degrees C. No differences were observed between the growth rates of axenic amastigotes and promastigotes. Flow cytometry-derived DNA histograms of axenic amastigotes and promastigotes were typical of exponentially growing cell populations. Moreover, axenic amastigotes were metabolically active as evidenced by the release of an immunoprecipitable extracellular acid phosphatase (SAcP) into their culture supernatant. Cell transformation was confirmed by transmission electronmicroscopic examination of thin sections and extended by fracture-flip survey which allowed differentiation of cell membranes. The ultrastructure and nanoanatomy of axenic amastigotes was identical to that of intracellular amastigotes. The production of large amounts of heat-shock axenic amastigotes suitable for biochemical and biological studies of differentiation in Leishmania donovani may have important implications in the development of prevention and/or treatment strategies.
杜氏利什曼原虫前鞭毛体经过热处理后形成无细胞内共生体的无鞭毛体阶段，并在37℃下进行长期培养。无细胞内共生体的无鞭毛体和前鞭毛体的生长速率未观察到差异。无细胞内共生体的无鞭毛体和前鞭毛体的流式细胞术衍生DNA直方图是典型的指数生长细胞群体。此外，无细胞内共生体的无鞭毛体具有代谢活性，这可通过可免疫沉淀的细胞外酸性磷酸酶（SAcP）释放到其培养上清液中得到证明。通过透射电子显微镜对薄片进行检查证实了细胞转化，并通过断裂-翻转调查进行了扩展，该调查允许区分细胞膜。无细胞内共生体的无鞭毛体的超微结构和纳米解剖结构与细胞内无鞭毛体相同。大量适合杜氏利什曼原虫分化生化和生物学研究的热休克无细胞内共生体无鞭毛体的产生可能对预防和/或治疗策略的发展具有重要意义。
