Generation of Leishmania donovani axenic amastigotes:: their growth and biological characteristics
Generation of Leishmania donovani axenic amastigotes:: their growth and biological characteristics
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DOI：
10.1016/j.ijpara.2003.10.011
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发表时间：
2004-02-01
影响因子：
4
通讯作者：
Dwyer, DM
中科院分区：
文献类型：
--
作者：
Debrabant, A;Joshi, MB;Dwyer, DM
在这份报告中，我们描述了一种从杜氏利什曼原虫1S-CL2D品系中产生和繁殖无菌无鞭毛体的体外培养系统。对这些体外培养的无鞭毛体的精细结构分析表明，它们具有杜氏乳杆菌组织源无鞭毛体的形态特征。此外，这些无性无鞭毛体(LdAxAm)被证明能够合成和释放一种与细胞内无鞭毛体产生的分泌型酸性磷酸酶类似的同工酶。这种LdAxAm也表达了与组织来源的无鞭毛体相似的表膜3‘-核苷酸酶活性。此外，与前鞭毛体相比，LdAxAm表达了显著水平的无鞭毛体特异性A2蛋白。此外，长期培养的LdAxAm对体外培养的人巨噬细胞和体内的金黄地鼠均有明显的感染性。因此，本文描述的体外培养系统为杜诺瓦尼乳杆菌1S-CL2D无性无鞭毛体的大量统一群体的产生提供了有用的工具。这种材料的提供将极大地促进关于这种寄生虫发育阶段的细胞和分子生物学的研究。(C)2003澳大利亚寄生虫学会有限公司出版。保留所有权利。
In this report, we describe an in vitro culture system for the generation and propagation of axenic amastigotes from the well characterised 1S-CL2D line of Leishmania donovani. Fine structure analyses of these in vitro-grown amastigotes demonstrated that they possessed morphological features characteristic of L. donovani tissue-derived amastigotes. Further, these axenic amastigotes (LdAxAm) were shown to synthesise and release a secretory acid phosphatase isoform similar to that produced by intracellular amastigotes. Such LdAxAm also expressed surface membrane 3'-nucleotidase enzyme activity similar to that of tissue-derived amastigotes. Moreover, LdAxAm, in contrast to promastigotes, expressed significant levels of the amastigote-specific A2 proteins. In addition, LdAxAm, derived from long term cultures of Ld 1S-CL2D promastigotes, had significant infectivity for both human macrophages in vitro and for hamsters in vivo. Thus, the in vitro culture system described herein provides a useful tool for the generation of large quantities of uniform populations of axenic amastigotes of the L. donovani 1S-CL2D line. The availability of such material should greatly facilitate studies concerning the cell and molecular biology of this parasite developmental stage. (C) 2003 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.