Leishmania donovani infection of macrophages results in quantitative and qualitative changes in the protein profile of extracellular vesicles (EVs) released by the infected host cells. We confirmed mass spectrometry results orthogonally by performing Western blots for several Leishmania-infected macrophage-enriched EVs (LieEVs) molecules. Several host cell proteins in LieEVs have been implicated in promoting vascular changes in other systems. We also identified 59 parasite-derived proteins in LieEVs, including a putative L. donovani homolog of mammalian vasohibins (LdVash), which in mammals promotes angiogenesis. We developed a transgenic parasite that expressed an endogenously tagged LdVash/mNeonGreen (mNG) and confirmed that LdVash/mNG is indeed expressed in infected macrophages and in LieEVs. We further observed that LieEVs induce endothelial cells to release angiogenesis promoting mediators including IL-8, G-CSF/CSF-3, and VEGF-A. In addition, LieEVs induce epithelial cell migration and tube formation by endothelial cells in surrogate angiogenesis assays. Taken together, these studies show that Leishmania infection alters the composition of EVs from infected cells and suggest that LieEVs may play a role in the promotion of vascularization of Leishmania infections.

中文翻译：

---

利什曼原虫感染的巨噬细胞释放可促进病变发展的细胞外囊泡。

巨噬细胞的利什曼原虫多诺万尼感染导致受感染宿主细胞释放的细胞外囊泡（EV）的蛋白质谱发生定量和定性变化。我们通过对几个感染利什曼原虫感染的巨噬细胞富集的电动汽车（LieEVs）分子进行蛋白质印迹来正交确认质谱结果。LieEV中的几种宿主细胞蛋白与促进其他系统中的血管变化有关。我们还发现了LieEV中的59种寄生虫衍生蛋白，包括推定的L. donovani哺乳动物血管抑制素（LdVash）的同系物，在哺乳动物中可促进血管生成。我们开发了一种转基因寄生虫，可表达内源标记的LdVash / mNeonGreen（mNG），并证实LdVash / mNG确实在感染的巨噬细胞和LieEV中表达。我们进一步观察到LieEVs诱导内皮细胞释放血管生成促进介体，包括IL-8，G-CSF / CSF-3和VEGF-A。另外，在替代性血管生成测定中，LieEVs通过内皮细胞诱导上皮细胞迁移和管形成。综上所述，这些研究表明利什曼原虫感染改变了感染细胞中电动车的组成，并表明LieEV可能在促进利什曼原虫感染的血管形成中起作用。