Abstract
A secreted killer toxin was detected through the cell wall ofPichia anomala cells by ultrastructural immunodetection with a specific monoclonal antibody (MAb KT4). MAb KT4 was successively detected by colloidal gold labeled streptavidin and biotinylated anti-mouse F(ab')2 antibodies. The antigenic determinants of the toxin were localized throughout the cytoplasm and the cell wall of killer yeast cells. The Lowicryl K4M-immunogold method gave very satisfactory results and showed that the killer toxin was somewhat concentrated in the yeast cell wall layers before being exported into the medium. In agreement with previous reports, the binding of MAb KT4 suggested that theP. anomala killer toxin secretion did not result from a homogeneous diffusion across the yeast cell wall.
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Abbreviations
- G15:
-
gold particles of 15 nm
- IEM:
-
immunoelectron microscopy
- IFA:
-
immunofluorescence assay
- MAb:
-
monoclonal antibody
- PBS:
-
phosphate buffered saline
- SAM/F(ab′)2 :
-
sheep antibodies anti-mouse F(ab′)2
- SBB:
-
Sabouraud buffered broth
References
Hernando FL, Cailliez JC, Trinel PA, Faille C, Mackenzie DWR, Poulain D. Qualitative and quantitative differences in recognition patterns ofCandida albicans protein and polysaccharide antigens by human sera. J Med Vet Mycol 1993; 31: 219–26.
Farley PC, Sheperd MG, Sullivan PA. The cellular location of proteases inCandida albicans. J Gen Microbiol 1986; 132: 3235–38.
Ferro-Novick S, Novick P, Field C, Scheckman R. Yeast secretory mutants that block the formation of active cell surface enzymes. J Cell Biol 1984; 98: 35–43.
Li R., Cutler JE. A cell surface/plasma membrane antigen ofCandida albicans. J Gen Microbiol 1991; 137: 455–64.
Andaluz E, Guillen A, Larriba G. Preliminary evidence for a glucan receptor in the yeastCandida albicans. Biochem J 1986; 240: 495–502.
Elorza MV, Rico H, Gozalbo D, Sentandreu R. Cell wall composition and protoplast regeneration inCandida albicans. Anton Leeuw 1983; 49: 457–69.
Cailliez JC, Poulain D. Les mécanismes de la glycosylation et leur importance dans le transport intracellulaire et la sécrétion des glycoprotéines des levures. J Mycol Med 1992; 2: 202–9.
Polonelli L, Morace G. Production and characterization of yeast killer toxin monoclonal antibodies. J Clin Microbiols 1986; 25: 460–62.
Cailliez JC, Gerloni M, Morace G, Conti S, Cantelli C, Polonelli L. Ultrastructural immunodetection of aPichia anomala killer toxin: a preliminary study. Biol Cell 1992; 75: 19–23.
Cailliez JC, Boudrissa A, Mackenzie DWR, Poulain D. Evaluation of a gold-silver staining method for detection and identification ofCandida species by light microscopy. Eur J Clin Microbiol 1991; 9: 886–91.
Cailliez JC, Poulain D. Analyse cytologique de l'expression d'un épitope porté par les glycoprotéines excrétées parCandida albicans. Ann Inst Past Microbiol 1988; 139: 171–88.
Hearn VM, Latgé JP, Prevost MC. Immunolocalization ofAspergillus fumigatus mycelial antigens. J Med Vet Mycol 1991; 29: 73–81.
Cailliez JC, Poulain D, Mackenzie DWR, Polonelli L. Cytological immunodetection of yeast glycoprotein secretion. Eur J Epidemiol 1992; 8: 452–59.
Poulain D, Cailliez JC, Dubremetz JF. Secretion of glycoproteins through the cell wall ofCandida albicans. Eur J Cell Biol 1989; 50: 94–99.
Cailliez JC, Cantelli C, Conti S, Gerloni M, Magliani W, Morace G, Polonelli L.Pichia anomala killer toxin secretion in the presence of tunicamycin. J Med Vet Mycol 1993; 31: 337–42.
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Cailliez, JC., Cantelli, C., Séguy, N. et al. Killer toxin secretion through the cell wall of the yeastPichia anomala . Mycopathologia 126, 173–177 (1994). https://doi.org/10.1007/BF01103772
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DOI: https://doi.org/10.1007/BF01103772