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The trypanosomatidsLeishmania amazonensis andTrypanosoma cruzi are excellent models for the study of the cell biology of intracellular protozoan infections. After their uptake by mammalian cells, the parasitic protozoan flagellatesL. amazonensis andT. cruzi lodge within acidified parasitophorous vacuoles (PVs). However, whereasL. amazonensis develops in spacious, phagolysosome-like PVs that may enclose numerous parasites,T. cruzi is transiently hosted within smaller vacuoles from which it soon escapes to the host cell cytosol. To investigate if parasite-specific vacuoles are required for the survival and differentiation ofT. cruzi , we constructed chimeric vacuoles by infection ofL. amazonensis amastigote-infected macrophages withT. cruzi epimastigotes (EPIs) or metacyclic trypomastigotes (MTs). These chimeric vacuoles, easily observed by microscopy, allowed the entry and fate ofT. cruzi inL. amazonensis PVs to be dynamically recorded by multidimensional imaging of coinfected cells. We found that althoughT. cruzi EPIs remained motile and conserved their morphology in chimeric vacuoles,T. cruzi MTs differentiated into amastigote-like forms capable of multiplying. These results demonstrate that the large adaptive vacuoles ofL. amazonensis are permissive toT. cruzi survival and differentiation and that noninfective EPIs are spared from destruction within the chimeric PVs. We conclude thatT. cruzi differentiation can take place inLeishmania -containing vacuoles, suggesting this occurs prior to their escape into the host cell cytosol.

Trypanosoma cruzi Differentiates and Multiplies within Chimeric Parasitophorous Vacuoles in Macrophages Coinfected with Leishmania amazonensis