title: Extracellular amastigotes of Trypanosoma cruzi are potent inducers of phagocytosis in mammalian cells.
abstract: The protozoan parasite Trypanosoma cruzi, the aetiological agent of Chagas' disease, has two infective life cycle stages, trypomastigotes and amastigotes. While trypomastigotes actively enter mammalian cells, highly infective extracellular amastigotes (type I T. cruzi) rely on actin-mediated uptake, which is generally inefficient in non-professional phagocytes. We found that extracellular amastigotes (EAs) of T. cruzi G strain (type I), but not Y strain (type II), were taken up 100-fold more efficiently than inert particles. Mammalian cell lines showed levels of parasite uptake comparable to macrophages, and extensive actin recruitment and polymerization was observed at the site of entry. EA uptake was not dependent on parasite-secreted molecules and required the same molecular machinery utilized by professional phagocytes during large particle phagocytosis. Transcriptional silencing of synaptotagmin VII and CD63 significantly inhibited EA internalization, demonstrating that delivery of supplemental lysosomal membrane to form the phagosome is involved in parasite uptake. Importantly, time-lapse live imaging using fluorescent reporters revealed phagosome-associated modulation of phosphoinositide metabolism during EA uptake that closely resembles what occurs during phagocytosis by macrophages. Collectively, our results demonstrate that T. cruzi EAs are potent inducers of phagocytosis in non-professional phagocytes, a process that may facilitate parasite persistence in infected hosts.
authors.affiliation: Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, USA.
authors.forename: Maria Cecilia
authors.initials: MC
authors.lastname: Fernandes
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Flannery
authors.forename: Norma
authors.initials: N
authors.lastname: Andrews
authors.forename: Renato A
authors.initials: RA
authors.lastname: Mortara
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D014355:Chagas Disease / Q000378:metabolism / Q000503:physiopathology*; D006367:HeLa Cells / Q000469:parasitology*; D006801:Humans; D008018:Life Cycle Stages / Q000502:physiology*; D050356:Lipid Metabolism / Q000502:physiology; D010587:Phagocytosis / Q000502:physiology*; D010716:Phosphatidylinositols / Q000378:metabolism; D050857:Synaptotagmins / Q000378:metabolism; D060149:Tetraspanin 30 / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000254:growth & development* / Q000472:pathogenicity
chemical_list: D000199:Actins; D010716:Phosphatidylinositols; D060149:Tetraspanin 30; D050857:Synaptotagmins