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Anti-VSG antibodies induce an increase in Trypanosoma evansi intracellular Ca2+ concentration

Published online by Cambridge University Press:  28 August 2008

M. MENDOZA ,
G. L. UZCANGA ,
R. PACHECO ,
H. ROJAS ,
L. M. CARRASQUEL ,
Y. GARCÍA-MARCHAN ,
X. SERRANO-MARTÍN ,
G. BENAÍM ,
J. BUBIS  and
A. MIJARES
M. MENDOZA*
Affiliation:
Centro de Estudios Biomédicos y Veterinarios, Universidad Nacional Experimental Simón Rodríguez, Caracas, Venezuela Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela
G. L. UZCANGA*
Affiliation:
Departamento de Biología Celular, Universidad Simón Bolívar, Caracas, Venezuela Centro de Biociencias y Medicina Molecular, Fundación Instituto de Estudios Avanzados – IDEA, Caracas, Venezuela
R. PACHECO
Affiliation:
Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela Departamento de Biología, Facultad Experimental de Ciencias y Tecnología, Universidad de Carabobo, Valencia, Venezuela
H. ROJAS
Affiliation:
Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela
L. M. CARRASQUEL
Affiliation:
Departamento de Biología Celular, Universidad Simón Bolívar, Caracas, Venezuela
Y. GARCÍA-MARCHAN
Affiliation:
Centro de Biociencias y Medicina Molecular, Fundación Instituto de Estudios Avanzados – IDEA, Caracas, Venezuela Instituto de Biología Experimental, Facultad de Ciencias, Universidad Central de Venezuela, Caracas, Venezuela
X. SERRANO-MARTÍN
Affiliation:
Centro de Biociencias y Medicina Molecular, Fundación Instituto de Estudios Avanzados – IDEA, Caracas, Venezuela Instituto de Biología Experimental, Facultad de Ciencias, Universidad Central de Venezuela, Caracas, Venezuela
G. BENAÍM
Affiliation:
Centro de Biociencias y Medicina Molecular, Fundación Instituto de Estudios Avanzados – IDEA, Caracas, Venezuela Instituto de Biología Experimental, Facultad de Ciencias, Universidad Central de Venezuela, Caracas, Venezuela
J. BUBIS
Affiliation:
Departamento de Biología Celular, Universidad Simón Bolívar, Caracas, Venezuela
A. MIJARES
Affiliation:
Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Caracas, Venezuela
*
*Corresponding authors: M. Mendoza, Centro de Estudios Biomédicos y Veterinarios, Instituto de Estudios Científicos y Tecnológicos, Universidad Nacional Experimental Simón Rodríguez, Apartado Postal 47925, Caracas, Venezuela. Tel: +58 212 5041853. Fax: +58 _12 5041093. E-mail: memendoza@cantv.net and G. L. Uzcanga, Centro de Biociencias y Medicina Molecular, Fundación Instituto de Estudios Avanzados – IDEA, Caracas, Venezuela. Tel: +58 212 9035112. Fax: +58 212 9035157. E-mail: guzcanga@idea.gob.ve
*Corresponding authors: M. Mendoza, Centro de Estudios Biomédicos y Veterinarios, Instituto de Estudios Científicos y Tecnológicos, Universidad Nacional Experimental Simón Rodríguez, Apartado Postal 47925, Caracas, Venezuela. Tel: +58 212 5041853. Fax: +58 _12 5041093. E-mail: memendoza@cantv.net and G. L. Uzcanga, Centro de Biociencias y Medicina Molecular, Fundación Instituto de Estudios Avanzados – IDEA, Caracas, Venezuela. Tel: +58 212 9035112. Fax: +58 212 9035157. E-mail: guzcanga@idea.gob.ve
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Summary

Trypanosoma evansi and Trypanosoma vivax have shown a very high immunological cross-reactivity. Anti-T. vivax antibodies were used to monitor changes in the T. evansi intracellular Ca2+ concentration ([Ca2+]i) by fluorometric ratio imaging from single parasites. A short-time exposure of T. evansi parasites to sera from T. vivax-infected bovines induced an increase in [Ca2+]i, which generated their complete lysis. The parasite [Ca2+]i boost was reduced but not eliminated in the absence of extracellular Ca2+ or following serum decomplementation. Decomplemented anti-T. evansi VSG antibodies also produced an increase in the parasite [Ca2+]i, in the presence of extracellular Ca2+. Furthermore, this Ca2+ signal was reduced following blockage with Ni2+ or in the absence of extracellular Ca2+, suggesting that this response was a combination of an influx of Ca2+ throughout membrane channels and a release of this ion from intracellular stores. The observed Ca2+ signal was specific since (i) it was completely eliminated following pre-incubation of the anti-VSG antibodies with the purified soluble VSG, and (ii) affinity-purified anti-VSG antibodies also generated an increase in [Ca2+]i by measurements on single cells or parasite populations. We also showed that an increase of the T. evansi [Ca2+]i by the calcium A-23187 ionophore led to VSG release from the parasite surface. In addition, in vivo immunofluorescence labelling revealed that anti-VSG antibodies induced the formation of raft patches of VSG on the parasite surface. This is the first study to identify a ligand that is coupled to calcium flux in salivarian trypanosomes.

Keywords

antibodiescalciumcomplementTrypanosoma evansiTrypanosoma vivaxvariant surface glycoproteins
Type
Original Articles
Information
Parasitology , Volume 135 , Issue 11 , September 2008 , pp. 1303 - 1315
Copyright
Copyright © 2008 Cambridge University Press

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