title: Cell invasion by Trypanosoma cruzi amastigotes of distinct infectivities: studies on signaling pathways.
abstract: Trypanosoma cruzi metacyclic trypomastigotes of the major phylogenetic lineages use specific signaling pathways to invade host cells. Using a panel of drugs, we studied if the differences in the ability of extracellular amastigotes (EA) from G (T. cruzi I) and CL (T. cruzi II) strains to invade host cells could be associated to activation of specific signaling routes. Sonicated extracts from G or CL strain EA induced transient raises in HeLa cell intracellular Ca(2+) levels in a dose-dependent manner. Treatment of EA with drugs that affect Ca(2+) release from inositol-1,4,5-triphosphate-sensitive stores did not significantly affect the infectivity of either strain, whereas EA of both strains treated with ionomycin plus NH(4)Cl or nigericin that release Ca(2+) from acidocalcisomes had their infectivity reduced. Treatment of parasites with adenylate cyclase activator forskolin increased the infectivity of both strains towards HeLa cells. These data, taken together, suggest that, for host cell invasion, G and CL strain EA engage signaling pathways that lead to an increase of cyclic adenosine monophosphate and Ca(2+) mobilization from acidocalcisomes. Moreover, treatment of EA with genistein reduced by approximately 45% the invasion of HeLa cells by G but not by CL strain, implicating a protein tyrosine kinase in the process. In line with this, HeLa cell extracts contained a protein tyrosine kinase activity that mediated the phosphorylation of 87- and 175-kDa polypeptides of EA from G but not from CL strain. Regarding the target cell response, the activation of host PI3 kinase appears to be required for invasion by either strain as treatment of HeLa cells with wortmannin reduced EA infectivity. These data overall reinforce the concept that cell invasion by T. cruzi EA markedly differs from the process involving metacyclic trypomastigotes.
authors.affiliation: Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo-UNIFESP, Escola Paulista de Medicina, Rua Botucatu, 862 6th Floor, 04023-062, São Paulo, Brazil.
authors.forename: Adriana B
authors.initials: AB
authors.lastname: Fernandes
authors.forename: Ivan
authors.initials: I
authors.lastname: Neira
authors.forename: Alice T
authors.initials: AT
authors.lastname: Ferreira
authors.forename: Renato A
authors.initials: RA
authors.lastname: Mortara
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D014355:Chagas Disease / Q000469:parasitology*; D002522:Chlorocebus aethiops; D000882:Haplorhini; D006367:HeLa Cells; D006801:Humans; D019869:Phosphatidylinositol 3-Kinases / Q000187:drug effects / Q000378:metabolism; D010802:Phylogeny; D015398:Signal Transduction; D014349:Trypanosoma cruzi / Q000145:classification / Q000472:pathogenicity*; D014709:Vero Cells; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D019869:Phosphatidylinositol 3-Kinases; D000077191:Wortmannin