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A Leishmania (L.) amazonensis ATP diphosphohydrolase isoform and potato apyrase share epitopes: antigenicity and correlation with disease progression

Published online by Cambridge University Press:  16 November 2007

E. S. COIMBRA
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil Departamento de Parasitologia, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil Departamento de Protozoologia, Laboratório de Imunomodulação, Instituto Oswaldo Cruz/FIOCRUZ – Rio de Janeiro, RJ, Brazil
S. C. GONÇALVES-DA-COSTA
Affiliation:
Departamento de Protozoologia, Laboratório de Imunomodulação, Instituto Oswaldo Cruz/FIOCRUZ – Rio de Janeiro, RJ, Brazil
B. L. S. COSTA
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
N. L. L. GIAROLA
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
F. A. REZENDE-SOARES
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
M. R. FESSEL
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
A. P. FERREIRA
Affiliation:
Departamento de Parasitologia, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
C. S. F. SOUZA
Affiliation:
Departamento de Protozoologia, Laboratório de Imunomodulação, Instituto Oswaldo Cruz/FIOCRUZ – Rio de Janeiro, RJ, Brazil
A. L. ABREU-SILVA
Affiliation:
Departamento de Patologia, Universidade Estadual do Maranhão, São Luís, Maranhão, Brazil Departamento de Protozoologia, Laboratório de Imunomodulação, Instituto Oswaldo Cruz/FIOCRUZ – Rio de Janeiro, RJ, Brazil
E. G. VASCONCELOS*
Affiliation:
Departamento de Bioquímica, Microbiologia e Imunologia, ICB, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil
*
*Corresponding author: Departamento de Bioquímica, ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, 36036-330, Juiz de Fora, MG, Brazil. Tel: +55 32 3229 3217. Fax: +55 32 3229 3216. E-mail: [email protected]

Summary

A Leishmania (Leishmania) amazonensis ATP diphosphohydrolase isoform was partially purified from plasma membrane of promastigotes by preparative non-denaturing polyacrylamide gel electrophoresis. SDS-PAGE followed by Western blots developed with polyclonal anti-potato apyrase antibodies identified diffuse bands of about 58–63 kDa, possibly glycosylated forms of this protein. By ELISA technique, a significantly higher total IgG antibody level against potato apyrase was found in serum from promastigote-infected mice, as compared to the uninfected mice, confirming both the existence of shared epitopes between the parasite and vegetable proteins, and the parasite ATP diphosphohydrolase antigenicity. By Western blotting, serum from amastigote-infected BALB/c mice recognizes both potato apyrase and this antigenic ATP diphosphohydrolase isoform isolated from promastigotes, suggesting that it is also expressed in the amastigote stage. The infection monitored along a 90-day period in amastigote-infected mice showed reactivity of IgG2a antibody in early steps of infection, while the disappearance of the IgG2a response and elevation of IgG1 antibody serum levels against that shared epitopes were associated with the progression of experimental leishmaniasis. This is the first observation of the antigenicity of a L. (L.) amazonensis ATP diphosphohydrolase isoform, and of the ability of cross-immunoreactivity with potato apyrase to differentiate serologically stages of leishmaniasis in infected mice.

Type
Original Articles
Copyright
Copyright © Cambridge University Press 2007

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