title: Lysosome biogenesis/scattering increases host cell susceptibility to invasion by Trypanosoma cruzi metacyclic forms and resistance to tissue culture trypomastigotes.
abstract: A fundamental question to be clarified concerning the host cell invasion by Trypanosoma cruzi is whether the insect-borne and mammalian-stage parasites use similar mechanisms for invasion. To address that question, we analysed the cell invasion capacity of metacyclic trypomastigotes (MT) and tissue culture trypomastigotes (TCT) under diverse conditions. Incubation of parasites for 1 h with HeLa cells in nutrient-deprived medium, a condition that triggered lysosome biogenesis and scattering, increased MT invasion and reduced TCT entry into cells. Sucrose-induced lysosome biogenesis increased HeLa cell susceptibility to MT and resistance to TCT. Treatment of cells with rapamycin, which inhibits mammalian target of rapamycin (mTOR), induced perinuclear lysosome accumulation and reduced MT invasion while augmenting TCT invasion. Metacylic trypomastigotes, but not TCT, induced mTOR dephosphorylation and the nuclear translocation of transcription factor EB (TFEB), a mTOR-associated lysosome biogenesis regulator. Lysosome biogenesis/scattering was stimulated upon HeLa cell interaction with MT but not with TCT. Recently, internalized MT, but not TCT, were surrounded by colocalized lysosome marker LAMP2 and mTOR. The recombinant gp82 protein, the MT-specific surface molecule that mediates invasion, induced mTOR dephosphorylation, nuclear TFEB translocation and lysosome biogenesis/scattering. Taken together, our data clearly indicate that MT invasion is mainly lysosome-dependent, whereas TCT entry is predominantly lysosome-independent.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, R. Pedro de Toledo, 669-6° andar, 04039-032, São Paulo, SP, Brazil.
authors.forename: Cristian
authors.initials: C
authors.lastname: Cortez
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, R. Pedro de Toledo, 669-6° andar, 04039-032, São Paulo, SP, Brazil.
authors.forename: Fernando
authors.identifier: http://orcid.org/0000-0002-5060-4334
authors.initials: F
authors.lastname: Real
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, R. Pedro de Toledo, 669-6° andar, 04039-032, São Paulo, SP, Brazil.
authors.forename: Nobuko
authors.initials: N
authors.lastname: Yoshida
mesh_terms: D000818:Animals; D014355:Chagas Disease / Q000235:genetics* / Q000469:parasitology; D004198:Disease Susceptibility / Q000378:metabolism / Q000469:parasitology; D006367:HeLa Cells; D054884:Host-Pathogen Interactions / Q000235:genetics*; D006801:Humans; D007303:Insect Vectors / Q000235:genetics / Q000469:parasitology / Q000472:pathogenicity; D052119:Lysosomal-Associated Membrane Protein 2 / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism / Q000469:parasitology*; D020123:Sirolimus / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D046509:Tissue Culture Techniques; D014349:Trypanosoma cruzi / Q000378:metabolism / Q000472:pathogenicity*
chemical_list: C497811:LAMP2 protein, human; D052119:Lysosomal-Associated Membrane Protein 2; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus
keywords: microbial-cell interaction; protozoa