Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina, UNIFESP, São Paulo, Brazil.
PLoS Negl Trop Dis. 2012;6(2):e1518. doi: 10.1371/journal.pntd.0001518. Epub 2012 Feb 14.
An important area in the cell biology of intracellular parasitism is the customization of parasitophorous vacuoles (PVs) by prokaryotic or eukaryotic intracellular microorganisms. We were curious to compare PV biogenesis in primary mouse bone marrow-derived macrophages exposed to carefully prepared amastigotes of either Leishmania major or L. amazonensis. While tight-fitting PVs are housing one or two L. major amastigotes, giant PVs are housing many L. amazonensis amastigotes. In this study, using multidimensional imaging of live cells, we compare and characterize the PV biogenesis/remodeling of macrophages i) hosting amastigotes of either L. major or L. amazonensis and ii) loaded with Lysotracker, a lysosomotropic fluorescent probe. Three dynamic features of Leishmania amastigote-hosting PVs are documented: they range from i) entry of Lysotracker transients within tight-fitting, fission-prone L. major amastigote-housing PVs; ii) the decrease in the number of macrophage acidic vesicles during the L. major PV fission or L. amazonensis PV enlargement; to iii) the L. amazonensis PV remodeling after homotypic fusion. The high content information of multidimensional images allowed the updating of our understanding of the Leishmania species-specific differences in PV biogenesis/remodeling and could be useful for the study of other intracellular microorganisms.
细胞内寄生生物学中的一个重要领域是原核或真核细胞内微生物对寄生泡（PV）的定制。我们很好奇，想比较一下在精心制备的利什曼原虫或亚马逊利什曼原虫的无鞭毛体刺激下，原代小鼠骨髓来源的巨噬细胞中 PV 的生物发生情况。虽然紧密贴合的 PV 中仅容纳一个或两个利什曼原虫无鞭毛体，但巨大的 PV 中容纳了许多亚马逊利什曼原虫无鞭毛体。在这项研究中，我们使用活细胞的多维成像，比较和描述了巨噬细胞中 PV 的生物发生/重塑：i）宿主利什曼原虫或亚马逊利什曼原虫的无鞭毛体，和 ii）负载溶酶体追踪剂，一种溶酶体亲脂性荧光探针。记录了三种利什曼原虫无鞭毛体寄生泡的动态特征：它们从 i）溶酶体追踪剂瞬态进入紧密贴合、易裂变的利什曼原虫无鞭毛体寄生泡；ii）在利什曼原虫 PV 裂变或亚马逊利什曼原虫 PV 扩大过程中，巨噬细胞酸性囊泡数量减少；到 iii）利什曼原虫 PV 重塑后同源融合。多维图像的高信息量更新了我们对利什曼原虫种间在 PV 生物发生/重塑方面差异的理解，并可用于研究其他细胞内微生物。
