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CrATP interferes in the promastigote-macrophage interaction in Leishmania amazonensis infection

Published online by Cambridge University Press:  17 June 2011

V. ENNES-VIDAL ,
R. O. S. CASTRO ,
C. BRITTO ,
H. BARRABIN ,
C. M. D'AVILA-LEVY  and
O. C. MOREIRA
V. ENNES-VIDAL
Affiliation:
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brazil
R. O. S. CASTRO
Affiliation:
Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
C. BRITTO
Affiliation:
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brazil
H. BARRABIN
Affiliation:
Laboratório de Transporte Iônico em Membranas Biológicas, Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, RJ, Brazil
C. M. D'AVILA-LEVY
Affiliation:
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brazil
O. C. MOREIRA*
Affiliation:
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brazil Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
*
*Corresponding author: Instituto Oswaldo Cruz/FIOCRUZ, CEP: 21040-360, Rio de Janeiro, RJ, Brazil. Tel: +55 21 3865 8153. Fax: +55 21 2590-3495. E-mail: [email protected]
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Summary

Recent have shown the relationship between Ecto-Nucleoside-Triphosphate-Diphosphohydrolases (Ecto-NTPDases or ecto-nucleotidases) and virulence and infectivity in trypanosomatids. In this work, the inhibition of the ecto-ATPase activities and promastigote growth of Leishmania amazonensis by CrATP was characterized. Furthermore, this compound was used to investigate the role of ecto-nucleotidase in the interaction of L. amazonensis with resident peritoneal macrophages obtained from BALB/c mice. CrATP partially inhibits the ecto-ATPase activity, presenting Ki values of 575·7±199·1 and 383·5±79·0 μm, in the presence or absence of 5 mm MgCl2, respectively. The apparent Kms for ATP (2·9±0·5 mm to Mg2+-dependent ecto-ATPase and 0·4±0·2 mm to Mg2+-independent ecto-ATPase activities) are not significantly altered by CrATP, suggesting a reversible non-competitive inhibition of both enzymes. When CrATP was added to the cultivation medium at 500 μm, it drastically inhibited the cellular growth. The interaction of promastigote forms of L. amazonensis with BALB/c peritoneal macrophages is strongly affected by CrATP. When the parasites were treated with 500 μm CrATP before interacting with macrophages, the adhesion and endocytic indices were strongly reduced to 53·0±14·8% and 39·8±1·1%, respectively. These results indicate that ecto-nucleotidase plays an important role in the infection process caused by Leishmania amazonensis.

Keywords

ecto-NTPDaseCrATPLeishmania amazonensisinfectivity
Type
Research Article
Information
Parasitology , Volume 138 , Issue 8 , July 2011 , pp. 960 - 968
Copyright
Copyright © Cambridge University Press 2011

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