title: Decoding the anti-Trypanosoma cruzi action of HIV peptidase inhibitors using epimastigotes as a model.
abstract: BACKGROUND
  Aspartic peptidase inhibitors have shown antimicrobial action against distinct microorganisms. Due to an increase in the occurrence of Chagas' disease/AIDS co-infection, we decided to explore the effects of HIV aspartic peptidase inhibitors (HIV-PIs) on Trypanosoma cruzi, the etiologic agent of Chagas' disease.
  
  
  RESULTS
  HIV-PIs presented an anti-proliferative action on epimastigotes of T. cruzi clone Dm28c, with IC50 values ranging from 0.6 to 14 µM. The most effective inhibitors, ritonavir, lopinavir and nelfinavir, also had an anti-proliferative effect against different phylogenetic T. cruzi strains. The HIV-PIs induced some morphological alterations in clone Dm28c epimastigotes, as reduced cell size and swollen of the cellular body. Transmission electron microscopy revealed that the flagellar membrane, mitochondrion and reservosomes are the main targets of HIV-PIs in T. cruzi epimastigotes. Curiously, an increase in the epimastigote-into-trypomastigote differentiation process of clone Dm28c was observed, with many of these parasites presenting morphological alterations including the detachment of flagellum from the cell body. The pre-treatment with the most effective HIV-PIs drastically reduced the interaction process between epimastigotes and the invertebrate vector Rhodnius prolixus. It was also noted that HIV-PIs induced an increase in the expression of gp63-like and calpain-related molecules, and decreased the cruzipain expression in epimastigotes as judged by flow cytometry and immunoblotting assays. The hydrolysis of a cathepsin D fluorogenic substrate was inhibited by all HIV-PIs in a dose-dependent manner, showing that the aspartic peptidase could be a possible target to these drugs. Additionally, we verified that ritonavir, lopinavir and nelfinavir reduced drastically the viability of clone Dm28c trypomastigotes, causing many morphological damages.
  
  
  CONCLUSIONS
  The results contribute to understand the possible role of aspartic peptidases in T. cruzi physiology, adding new in vitro insights into the possibility of exploiting the use of HIV-PIs in the clinically relevant forms of the parasite.
authors.affiliation: Laboratório de Investigação de Peptidases, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes (IMPG), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Leandro S
authors.initials: LS
authors.lastname: Sangenito
authors.affiliation: Laboratório de Biologia Celular, Instituto Oswaldo Cruz (IOC), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, Brazil.
authors.forename: Rubem F S
authors.initials: RF
authors.lastname: Menna-Barreto
authors.affiliation: Laboratório de Biologia Molecular e Doenças Endêmicas, IOC, FIOCRUZ, Rio de Janeiro, Brazil.
authors.forename: Claudia M
authors.initials: CM
authors.lastname: D Avila-Levy
authors.affiliation: Laboratório de Investigação de Peptidases, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes (IMPG), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil; Programa de Pós-Graduação em Bioquímica, Instituto de Química, UFRJ, Rio de Janeiro, Brazil.
authors.forename: André L S
authors.initials: AL
authors.lastname: Santos
authors.affiliation: Laboratório de Investigação de Peptidases, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes (IMPG), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.
authors.forename: Marta H
authors.initials: MH
authors.lastname: Branquinha
mesh_terms: D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology*; D057055:Aspartic Acid Proteases / Q000037:antagonists & inhibitors / Q000378:metabolism; D002219:Carbamates / Q000494:pharmacology; D005663:Furans; D006790:Host-Parasite Interactions / Q000187:drug effects; D019469:Indinavir / Q000494:pharmacology; D007303:Insect Vectors / Q000469:parasitology; D061466:Lopinavir / Q000494:pharmacology; D046529:Microscopy, Electron, Transmission; D019888:Nelfinavir / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology*; D019438:Ritonavir / Q000494:pharmacology; D019258:Saquinavir / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000187:drug effects* / Q000254:growth & development / Q000648:ultrastructure
chemical_list: D019380:Anti-HIV Agents; D002219:Carbamates; D005663:Furans; D011480:Protease Inhibitors; D013449:Sulfonamides; D014344:Trypanocidal Agents; D061466:Lopinavir; C095108:amprenavir; D019469:Indinavir; D057055:Aspartic Acid Proteases; D019888:Nelfinavir; D019258:Saquinavir; D019438:Ritonavir