Analyse cytologique de l'expression d'un épitope porté par les glycoprotéines excrétées par Candida albicans

Abstract

The present study concerns an epitope identified by a monoclonal IgM, named 5B2, generated against the parasitic phase of Candida albicans. The epitope was previously shown to be carried by excreted C. albicans glycoproteins and to be present in the sera of patients suffering from systemic candidiasis. The cytological analysis of the epitope expression was investigated in 3 different yeast strains: the C. albicans strain from which 5B2 was generated (VW.32); a C. albicans mutant, deficient in cell wall mannans (KD.102); and a Sacchromyces cerevisiae strain. Immunofluorescence assays using IgM-5B2 showed discontinoous labelling with the VW.32 strain and no labelling with the 2 other yeast strains; however, the superficial structures of the 3 strains reacted homogeneously with ConA. Ultrastructural immunodetection experiments performed with the VW.32 cells, using gold-conjugated monoclonal antibody, revealed the presence of the epitope in the vacuolo-vesicular system, the periphery of the cytoplasm, the periplasmic space and the cell wall. Under the same conditions, cells from the KD.102 strain only exhibited weak cytoplasmic labelling whereas the presence of the epitope in S. cerevisiae blastoconidia was restricted to the vesicles. Competition and double labelling experiments with IgM and ConA showed that the epitope, distributed on the great majority of VW.32 glycoproteins, is shared by a lesser proportion of the KD.102 glycoproteins and only by some vesicular glycoproteins of S. cerevisiae. Inhibition of the N-glycosylation process of the VW.32 strain by tunicamycin resulted in the absence of cytokinesis and germ tube formation. In such cells, epitope 5B2 was no longer expressed on the bud surface. These cytological results concerning the C. albicans epitope are discussed in relation to recent, more general biochemical data on the yeast glycosylation process.