Chaves C S, Soares D C, Da Silva R P, Saraiva E M
Departamento de Imunologia, Instituto de Microbiologia-UFRJ, Rio de Janeiro, RJ, Brazil.
Exp Parasitol. 2003 Mar-Apr;103(3-4):152-9. doi: 10.1016/s0014-4894(03)00098-5.
Leishmania metacyclogenesis is associated with changes in morphology, gene expression, and structural alterations of the lipophosphoglycan (LPG), the promastigote most abundant surface glycolipid. Purification of metacyclics is accomplished using lectins or monoclonal antibodies (MAbs) that exploit stage-specific differences in the LPG. Besides, LPG displays extensive interspecies polymorphisms and is synthesized by promastigotes of all species investigated to date. In this work we studied the species- and stage-specificity of two MAbs (3A1-La and LuCa-D5) used to purify metacyclics of Leishmania amazonensis. Their ability to recognize different members of the Trypanosomatidae family was tested by direct agglutination, indirect immunofluorescence, and dot-blot analysis of LPG. We found that both MAbs were highly selective for L. amazonensis: 3A1-La recognized only promastigotes and LuCa-D5 labeled amastigote and promastigote stages of this species. These MAbs might be useful for Leishmania typing.
利什曼原虫的循环前体形成与形态变化、基因表达以及脂磷壁酸(LPG)的结构改变有关,LPG是前鞭毛体中最丰富的表面糖脂。利用凝集素或单克隆抗体(MAb)来纯化循环前体,这些凝集素或单克隆抗体利用了LPG中的阶段特异性差异。此外,LPG表现出广泛的种间多态性,并且由迄今为止研究的所有物种的前鞭毛体合成。在这项工作中,我们研究了用于纯化亚马逊利什曼原虫循环前体的两种单克隆抗体(3A1-La和LuCa-D5)的种属和阶段特异性。通过直接凝集、间接免疫荧光和LPG的斑点印迹分析,测试了它们识别锥虫科不同成员的能力。我们发现这两种单克隆抗体对亚马逊利什曼原虫具有高度选择性:3A1-La仅识别前鞭毛体,而LuCa-D5标记该物种的无鞭毛体和前鞭毛体阶段。这些单克隆抗体可能对利什曼原虫分型有用。