Cruzipain Activates Latent TGF-β from Host Cells during T. cruzi Invasion.
Several studies indicate that the activity of cruzipain, the main lysosomal cysteine peptidase of Trypanosoma cruzi, contributes to parasite infectivity. In addition, the parasitic invasion process of mammalian host cells is described to be dependent on the activation of the host TGF-β signaling pathway by T. cruzi. Here, we tested the hypothesis that cruzipain could be an important activator of latent TGF-β and thereby trigger TGF-β-mediated events crucial for the development of Chagas disease. We found that live epimastigotes of T. cruzi, parasite lysates and purified cruzipain were able to activate latent TGF-β in vitro. This activation could be inhibited by the cysteine peptidase inhibitor Z-Phe-Ala-FMK. Moreover, transfected parasites overexpressing chagasin, a potent endogenous cruzipain inhibitor, prevented latent TGF-β activation. We also observed that T. cruzi invasion, as well as parasite intracellular growth, were inhibited by the administration of Z-Phe-Ala-FMK or anti-TGF-β neutralizing antibody to Vero cell cultures. We further demonstrated that addition of purified cruzipain enhanced the invasive activity of trypomastigotes and that this effect could be completely inhibited by addition of a neutralizing anti-TGF-β antibody. Taken together, these results demonstrate that the activities of cruzipain and TGF-β in the process of cell invasion are functionally linked. Our data suggest that cruzipain inhibition is an interesting chemotherapeutic approach for Chagas disease not only because of its trypanocidal activity, but also due to the inhibitory effect on TGF-β activation.
Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil; Laboratório de Investigação Cardiovascular, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
Patrícia Mello
PM
Ferrão
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
Claudia Masini
CM
d'Avila-Levy
Laboratório de Inovações em Terapias, Ensino e Bioprodutos, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
Tania Cremonini
TC
Araujo-Jorge
Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
Wim Maurits
WM
Degrave
Laboratório de Biotecnologia e Fisiologia de Infecções Virais, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
Antônio da Silva
Ada S
Gonçalves
Laboratório de Investigação Cardiovascular, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil; Programa Integrado de doença de Chagas, Fiocruz, Rio de Janeiro, Brazil.
Luciana Ribeiro
LR
Garzoni
Laboratório de Bioquímica e Biologia Molecular de Peptidases, Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Brazil.
Ana Paula
AP
Lima
INSERM, Unité 1036, Grenoble, F-38054, France; Université Grenoble-Alpes-Grenoble, F-38041, France; CEA, DSV,iRTSV, Laboratory of Biology of Cancer and Infection, Grenoble, F-38054, France.
Jean Jacques
JJ
Feige
INSERM, Unité 1036, Grenoble, F-38054, France; Université Grenoble-Alpes-Grenoble, F-38041, France; CEA, DSV,iRTSV, Laboratory of Biology of Cancer and Infection, Grenoble, F-38054, France.
Sabine
S
Bailly
Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil.
Leila
L
Mendonça-Lima
Laboratório de Genômica Funcional e Bioinformática, Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brazil; Programa Integrado de doença de Chagas, Fiocruz, Rio de Janeiro, Brazil.
Mariana Caldas
MC
Waghabi
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002522:Chlorocebus aethiops; D003546:Cysteine Endopeptidases / Q000494:pharmacology*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004151:Dipeptides; D006790:Host-Parasite Interactions / Q000187:drug effects; D007659:Ketones; D010271:Parasites / Q000187:drug effects; D015800:Protozoan Proteins / Q000494:pharmacology; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000276:immunology / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000254:growth & development / Q000502:physiology*; D014709:Vero Cells
D000906:Antibodies; D015853:Cysteine Proteinase Inhibitors; D004151:Dipeptides; D007659:Ketones; D015800:Protozoan Proteins; D016212:Transforming Growth Factor beta; C435145:chagasin, Trypanosoma; C057383:MDL 201053; D003546:Cysteine Endopeptidases; C063214:cruzipain