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Characterization of a Leishmania antigen associated with cytoplasmic vesicles resembling endosomal-like structure

Published online by Cambridge University Press:  06 April 2009

B. Yahiaoui ,
M. Loyens ,
A. Taibi ,
R. Schöneck ,
J. F. Dubremetz  and
M. A. Ouaissi
B. Yahiaoui
Affiliation:
Research Laboratory on Trypanosomatids, INSERM U 167, Institut Pasteur, Lille, France
M. Loyens
Affiliation:
Research Laboratory on Trypanosomatids, INSERM U 167, Institut Pasteur, Lille, France
A. Taibi
Affiliation:
Research Laboratory on Trypanosomatids, INSERM U 167, Institut Pasteur, Lille, France
R. Schöneck
Affiliation:
Research Laboratory on Trypanosomatids, INSERM U 167, Institut Pasteur, Lille, France
J. F. Dubremetz
Affiliation:
INSERM U42, Rue Jules Guesde, Domaine du Certia, Villeneuve d'ascq, Lille, France
M. A. Ouaissi
Affiliation:
Research Laboratory on Trypanosomatids, INSERM U 167, Institut Pasteur, Lille, France
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Summary

In the present study we have used antibodies to Leishmania major promastigote antigens which were eluted from a glutathione-agarose column (LmGbp) and could identify several parasite components among different Leishmania species by using immunoprecipitation and Western blot techniques. The results also showed that some of LmGbp are present among the molecules released into the culture medium. Moreover, immunofluorescence assays clearly demonstrated that LmGbp are expressed by intracellular amastigotes. The electron micrographs of thawed cryosections of L. major-infected cells revealed that the antigens were associated with the membrane of the phagocytic vacuole. Moreover, the Western blot technique allowed us to identify, using other Leishmania species extracts and anti-LmGbp antibodies, a major polypeptide of an apparent molecular mass of 66 kDa. Immunofluorescence studies suggested that the 66 kDa polypeptide is associated with intracytoplasmic vesicles. Cryosections of Leishmania promastigotes improved the fine structure preservation of the organelles and enabled a number of features to be seen, particularly the structures considered as vesicles, which appeared as a complex tubulo-vesicular structure resembling mammalian cell endosomes and Leishmania organelles previously named ‘megasomes’. Further studies using antibodies against the native 66 kDa protein will be needed to investigate the localization of the protein at the ultrastructural level and to follow its intracellular vesicular traffic.

Keywords

Leishmaniaimmunofluorescenceimmunolocalizationendosomal-like structure
Type
Research Article
Information
Parasitology , Volume 107 , Issue 5 , December 1993 , pp. 497 - 507
Copyright
Copyright © Cambridge University Press 1993

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