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Dissimilar peptidase production by avirulent and virulent promastigotes of Leishmania braziliensis: inference on the parasite proliferation and interaction with macrophages

Published online by Cambridge University Press:  27 July 2009

A. K. C. LIMA ,
C. G. R. ELIAS ,
J. E. O. SOUZA ,
A. L. S. SANTOS  and
P. M. L. DUTRA
A. K. C. LIMA
Affiliation:
Disciplina de Parasitologia, Departamento de Microbiologia, Imunologia e Parasitologia (DMIP), Faculdade de Ciências Médicas (FCM), Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, Brazil
C. G. R. ELIAS
Affiliation:
Laboratório de Estudos Integrados em Bioquímica Microbiana, Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
J. E. O. SOUZA
Affiliation:
Disciplina de Parasitologia, Departamento de Microbiologia, Imunologia e Parasitologia (DMIP), Faculdade de Ciências Médicas (FCM), Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, Brazil
A. L. S. SANTOS
Affiliation:
Laboratório de Estudos Integrados em Bioquímica Microbiana, Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
P. M. L. DUTRA*
Affiliation:
Disciplina de Parasitologia, Departamento de Microbiologia, Imunologia e Parasitologia (DMIP), Faculdade de Ciências Médicas (FCM), Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, Brazil
*
*Corresponding author: Departamento de Microbiologia, Imunologia e Parasitologia, Rua Prof. Manoel de Abreu, 444, PAPC 5° andar, Vila Isabel, Rio de Janeiro, RJ 20550-170, Brazil. Tel: +55 21 2587 6487. Fax: +55 21 2587 6148. E-mail: [email protected]
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Summary

In the present paper, we have analysed the cellular and extracellular proteolytic activity profiles in 2 distinct Leishmania braziliensis strains: a recently isolated (virulent) and a laboratory-adapted (avirulent) strain. Quantitative and qualitative differences on the peptidase expression were observed in both strains. For instance, low-molecular mass acidic cysteine peptidase activities were detected exclusively in the virulent strain. Similarly, metallopeptidase activities were mainly produced by L. braziliensis virulent promastigotes. Interestingly, metallo- and cysteine peptidase activities were drastically reduced after several in vitro passages of the virulent strain. Western blotting, flow cytometry and fluorescence microscopy analyses were performed to detect homologous of the major leishmania metallopeptidase (gp63) and cysteine peptidase (cpb) in virulent and avirulent strains of L. braziliensis. Our results revealed that the virulent strain produced higher amounts of gp63 and cpb molecules, detected both in the surface and cytoplasm regions, than the avirulent counterpart. Metallo- (1,10-phenanthroline and EGTA) and cysteine peptidase (E-64) inhibitors arrested the growth of L. braziliensis virulent strain in a dose-dependent manner, as well as the association index with peritoneal murine macrophages. Conversely, these peptidase inhibitors did not affect either the proliferation or the cellular interaction of the avirulent strain. Corroborating these findings, the pre-treatment of the virulent strain with both anti-peptidase antibodies promoted a prominent reduction in the interaction with macrophages, while the association index of the avirulent strain to macrophage was only slightly diminished. Moreover, the spent culture medium from virulent strain significantly enhanced the association index between avirulent strain and macrophages, and this effect was reversed by 1,10-phenanthroline. Collectively, the results presented herein suggest that peptidases participate in several crucial processes of L. braziliensis.

Keywords

Leishmania braziliensispeptidasesproteolytic inhibitorsgrowthcellular interactiongp63cpb
Type
Research Article
Information
Parasitology , Volume 136 , Issue 10 , September 2009 , pp. 1179 - 1191
Copyright
Copyright © Cambridge University Press 2009

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