Abstract
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.
Highlights
Leishmania spp. are dimorphic trypanosomatid parasites that alternate between extracellular promastigote forms found in insect vectors and intracellular amastigote forms found in mammalian hosts
Using multidimensional live cell imaging, we examined the biogenesis of the two parasitophorous vacuoles (PVs) phenotypes in macrophages exposed to L. major or L. amazonensis amastigotes
The biogenesis/remodeling of the two types of Leishmania PVs is accompanied by a reduction in the number of macrophage acidic vesicles
Summary
Leishmania spp. are dimorphic trypanosomatid parasites that alternate between extracellular promastigote forms found in insect vectors and intracellular amastigote forms found in mammalian hosts. The large PVs that shelter parasites of the L. mexicana complex, such as L. amazonensis, can contain numerous amastigotes, often bound by their posterior poles to the internal face of the PVs [4] The biogenesis of these two types of PVs involves the acquisition of host cell late endosomes membrane markers, as shown in infected cells immunostained for lysosomeassociated membrane proteins (LAMPs), Rab GTPases, cathepsin, proton ATPases, and MHC class II molecules [2,5,6,7,8]. The acquisition of these markers is a coordinated event that results in a ‘‘mature’’ PV, which is presumably required for the survival and multiplication of the parasites
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