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Leishmania mexicana: promastigotes and amastigotes secrete protein phosphatases and this correlates with the production of inflammatory cytokines in macrophages

Published online by Cambridge University Press:  25 May 2016

A. R. ESCALONA-MONTAÑO ,
D. M. ORTIZ-LOZANO ,
A. ROJAS-BERNABÉ ,
A. A. WILKINS-RODRIGUEZ ,
H. TORRES-GUERRERO ,
R. MONDRAGÓN-FLORES ,
R. MONDRAGÓN-GONZALEZ ,
I. BECKER ,
L. GUTIÉRREZ-KOBEH  and
M. M. AGUIRRE-GARCIA
A. R. ESCALONA-MONTAÑO
Affiliation:
Facultad de Medicina, Unidad de Investigación en Medicina Experimental, Universidad Nacional Autónoma de México, México, DF, México
D. M. ORTIZ-LOZANO
Affiliation:
Facultad de Medicina, Unidad de Investigación en Medicina Experimental, Universidad Nacional Autónoma de México, México, DF, México
A. ROJAS-BERNABÉ
Affiliation:
Facultad de Medicina, Unidad de Investigación en Medicina Experimental, Universidad Nacional Autónoma de México, México, DF, México
A. A. WILKINS-RODRIGUEZ
Affiliation:
Facultad de Medicina, Unidad de Investigación en Medicina Experimental, Universidad Nacional Autónoma de México, México, DF, México
H. TORRES-GUERRERO
Affiliation:
Facultad de Medicina, Unidad de Investigación en Medicina Experimental, Universidad Nacional Autónoma de México, México, DF, México
R. MONDRAGÓN-FLORES
Affiliation:
Departamento de Bioquímica, Centro de Investigación y Estudios Avanzados (CINVESTAV-IPN), Av. Instituto Politécnico Nacional No. 2508, Col. San Pedro Zacatenco, México, DF, México
R. MONDRAGÓN-GONZALEZ
Affiliation:
Departamento de Genética y Biología Molecular, Centro de Investigación y Estudios Avanzados (CINVESTAV-IPN), Av. Instituto Politécnico Nacional No. 2508, Col. San Pedro Zacatenco, México, DF, México
I. BECKER
Affiliation:
Facultad de Medicina, Unidad de Investigación en Medicina Experimental, Universidad Nacional Autónoma de México, México, DF, México
L. GUTIÉRREZ-KOBEH
Affiliation:
Facultad de Medicina, Unidad de Investigación en Medicina Experimental, Universidad Nacional Autónoma de México, México, DF, México
M. M. AGUIRRE-GARCIA*
Affiliation:
Facultad de Medicina, Unidad de Investigación en Medicina Experimental, Universidad Nacional Autónoma de México, México, DF, México
*
*Corresponding author: Facultad de Medicina, Unidad de Investigación en Medicina Experimental, UNAM, Dr. Balmis 148, Colonia Doctores, México, DF 06726, México. E-mail: maguirre@unam.mx
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Summary

Phosphatase activity of Leishmania spp. has been shown to deregulate the signalling pathways of the host cell. We here show that Leishmania mexicana promastigotes and amastigotes secrete proteins with phosphatase activity to the culture medium, which was higher in the Promastigote Secretion Medium (PSM) as compared with the Amastigote Secretion Medium (ASM) and was not due to cell lysis, since parasite viability was not affected by the secretion process. The biochemical characterization showed that the phosphatase activity present in PSM was higher in dephosphorylating the peptide END (pY) INASL as compared with the peptide RRA (pT)VA. In contrast, the phosphatase activity in ASM showed little dephosphorylating capacity for both peptides. Inhibition assays demonstrated that the phosphatase activity of both PSM and ASM was sensible only to protein tyrosine phosphatases inhibitors. An antibody against a protein phosphatase 2C (PP2C) of Leishmania major cross-reacted with a 44·9 kDa molecule in different cellular fractions of L. mexicana promastigotes and amastigotes, however, in PSM and ASM, the antibody recognized a protein about 70 kDa. By electron microscopy, the PP2C was localized in the flagellar pocket of amastigotes. PSM and ASM induced the production of tumor necrosis factor alpha, IL-1β, IL-12p70 and IL-10 in human macrophages.

Keywords

Leishmania mexicanaprotein tyrosine phosphataseprotein phosphatase PP2Cinflammatory cytokines
Type
Research Article
Information
Parasitology , Volume 143 , Issue 11 , September 2016 , pp. 1409 - 1420
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Copyright
Copyright © Cambridge University Press 2016 

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