1887

Abstract

IgG hybridomas were produced which preferentially reacted with cell-surface antigens of either yeast cells or hyphae of . Four mAbs were used in an immunostaining procedure to follow the expression dynamics of these antigens in media supplemented with glucose or galactose. Yeast cell growth was analysed during the lag phase, the early- and late-exponential phases and the stationary phase, and mycelium formation was analysed between 0.5 and 24 h induction at 37 C. It appears that yeast cell-surface antigens 5C11 and 2E11 are expressed throughout all phases of yeast cell growth as well as on young hyphae after up to 1 h induction. Longer hyphae only faintly react with these two mAbs as they switch to hyphal cell-surface antigens 2G8 and 4E1 after 3 h induction. The reactivity to mAbs 2G8 and 4E1 was induced after a 3 h temperature shift and was confined to the terminal third of growing mycelia. Growth and hyphae induction in galactose prolonged the reactivity of young hyphae with the two anti-yeast-cell mAbs, whereas the expression of surface antigens 2G8 and 2E11 appeared delayed and desynchronized on hyphae. Whereas a similar reactivity was found with ten ATCC strains of , four clinical isolates had a unique pattern of reactivity. Immunoblot analyses of DTT extracts of cell-surface constituents indicated that the antigens were proteinaceous in nature and showed that yeast-cell antigens 5C11 and 2E11 are detected in four bands between 68 and 104 kDa, whereas mycelial antigens 4E1 and 2G8 are detected in 117 kDa and 104 kDa bands found in mycelial but not in yeast-cell extracts. Present data support the concept of a dynamic balance in the expression of phase-specific antigens in .

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1996-05-01
2021-07-31
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References

  1. Ansorge W. Fast and sensitive detection of protein and DNA bands by treatment with potassium permanganate. J Biochem Biophys Methods 1985; 11:13–20
    [Google Scholar]
  2. Bazin R., Lemieux R. Increased proportion of B-cell hybridomas secreting monoclonal antibodies of desired specificity in cultures containing macrophage-derived hybridoma growth factor (IL-6). J Immunol Methods 1989; 116:245–249
    [Google Scholar]
  3. Brawner D.L., Cutler J.E. Variability in expression of cell surface antigens of Candida albicans during morphogenesis. Infect Immun 1986; 51:337–343
    [Google Scholar]
  4. Casanova M., Gil M.L., Cardenoso L., Martinez J.P., Sentandreu R. Identification of wall-specific antigens synthesized during germ tube formation by Candida albicans. Infect Immun 1989; 57:262–271
    [Google Scholar]
  5. Cassone A., Torosantucci A., Boccanera M., Pellegreni G., Palma G., Malavasi F. Production and characterization of a monoclonal antibody to a cell surface glucomannoprotein constituent of Candida albicans and other pathogenic Candida species. J Med Microbiol 1988; 27:233–238
    [Google Scholar]
  6. Chaffin W.L., Skudlarek J., 8« Morrow K.J. Variable expression of a surface determinant during proliferation of Candida albicans. Infect Immun 1988; 56:302–309
    [Google Scholar]
  7. Chardes T., Piecharzyk M., Cavailles V., Salhi S.L., Pau B., Bastide J.M. Production and partial characterization of 'snû-Candida monoclonal antibodies. Ann Inst Pasteur Immunol 1986; 137C:117–125
    [Google Scholar]
  8. Critchley I.A., Douglas L.J. Isolation and characterization of an adhesin from Candida albicans. J Gen Microbiol 1987; 133:629–636
    [Google Scholar]
  9. Cutler J.E. Putative virulence factors of Candida albicans. Annu Rev Microbiol 1991; 45:187–218
    [Google Scholar]
  10. Garzón S., Marquis G., Montplaisir S., Kurstak E., Benhamou N. Antigenic structure of Candida albicans. In Immunology of Fungal Diseases 1989 Edited by Kurstak E. New York: Dekker; pp 3–36
    [Google Scholar]
  11. Gerard C. Purification of glycoproteins. Methods Enzymol 1990; 182:529–538
    [Google Scholar]
  12. Hasendever H.F., Mitchell W.O. Antigenic studies of Candida. I. Observations of two antigenic groups of Candida albicans. J Bacteriol 1961; 82:570–573
    [Google Scholar]
  13. Hasendever H.F., Mitchell W.O., Loewe J. Antigenic studies of Candida II Antigenic relation of Candida albicans group A and group B to Candida stellatoidea and Candida tropicalis. J Bacteriol 1961; 82:574–577
    [Google Scholar]
  14. Hopwood U., Poulain D., Fortier B., Evans G., Vernes A. A monoclonal antibody to a cell wall component of Candida albicans. Infect Immun 1986; 54:222–227
    [Google Scholar]
  15. Jones J.M. Humoral immune response to Candida albicans. In Immunology of Fungal Diseases 1989 Edited by Kurstak E. New York: Dekker; 47 pp 375–400
    [Google Scholar]
  16. Lacasse M., Fortier C., Trudel L., Collet A.J., Deslauriers N. Experimental oral candidosis in the mouse: microbiological and histological aspects. J Oral Pathol Med 1990; 19:136–141
    [Google Scholar]
  17. Lamy L., L£veille C., Fortier C., Lacasse M., Rojas M.P., Deslauriers N. In situ detection of Candida albicans using oral mucosal imprints. J Microbiol Methods 1992; 15:311–320
    [Google Scholar]
  18. Leusch H.G. Detection and characterization of two antigens specific for cell walls of Candida albicans mycelial growth phase. Curr Microbiol 1989; 19:193–198
    [Google Scholar]
  19. Li R.K., Cutler J.E. A cell surface/plasma membrane antigen of Candida albicans. J Gen Microbiol 1991; 137:455–464
    [Google Scholar]
  20. Linehan L., Wadsworth E., Calderone R. Candida albicans C3d receptor isolated by using a monoclonal antibody. Infect Immun 1988; 56:1981–1986
    [Google Scholar]
  21. Marot-Leblond A., Robert R., Aubry J., Ezcurra P., Senet J.M. Identification and immunochemical characterization of a germ tube specific antigen of Candida albicans. FFMS Immunol Med Microbiol 1993; 1:175–186
    [Google Scholar]
  22. McCourtie J., Douglas L.J. Relationship between cell surface composition, adherence and virulence of Candida albicans. Infect Immun 1984; 45:6–12
    [Google Scholar]
  23. McCourtie J., Douglas L.J. Extracellular polymer of Candida albicans', isolation, analysis and role in adhesion. J Gen Microbiol 1985; 131:495–503
    [Google Scholar]
  24. Miyakawa Y., Kagaya K., Fukazawa Y. Production and characterization of agglutinating monoclonal antibodies against predominant antigenic factors for Candida albicans. Infect Immun 1986; 23:881–886
    [Google Scholar]
  25. Odds F.C. Ecology of Candida and epidemiology of candidosis. In Candida and Candidosis 1988 Edited by Odds F.C. London: Bailliere Tindall; pp 69–92
    [Google Scholar]
  26. Ollert M.D., Calderone R.A. A monoclonal antibody that defines a surface antigen on Candida albicans hyphae cross-reacts with blastospore protoplasts. Infect Immun 1990; 58:625–631
    [Google Scholar]
  27. Ponton J., Jones J.M. Identification of two germ-tube-specific cell wall antigens of Candida albicans. Infect Immun 1985; 54:864–868
    [Google Scholar]
  28. Ponton J., Marot-Leblond A., Ezkurra P.A., Barturen B., Robert R., Senet J.M. Characterization of Candida albicans cell wall antigens with monoclonal antibodies. Infect Immun 1993; 61:4842–4847
    [Google Scholar]
  29. Poulain D., Hopwood U., Vernes A. Antigenic variability of Candida albicans. Crit Rev Microbiol 1985; 12:223–270
    [Google Scholar]
  30. Poulain D., Cailliez J.C., Dubremetz J.F. Secretion of glycoproteins through the cell wall of Candida albicans. Fur J Cell Biol 1989; 50:94–99
    [Google Scholar]
  31. Smail E.H., Jones J.M. Demonstration and solubilization of antigens expressed primarily on the surfaces of Candida albicans germ tubes. Infect Immun 1984; 45:74–81
    [Google Scholar]
  32. Stockbine N.A., Largen M.T., Buckley H.R. Production and characterization of three monoclonal antibodies to Candida albicans proteins. Infect Immun 1984; 43:1012–1018
    [Google Scholar]
  33. Summers D.F., Grollman A.P., Hasenclever H.F. Polysaccharide antigen of the Candida cell wall. I Immunol 1964; 92:491–499
    [Google Scholar]
  34. Sundstrom P.M., Kenny G.E. Characterization of antigen specific to the surface of germ tubes of Candida albicans by immunofluorescence. Infect Immun 1984; 43:850–855
    [Google Scholar]
  35. Sundstrom P.M., Kenny G.E. Enzymatic release of germ tube-specific antigens from cell walls of Candida albicans. Infect Immun 1985; 49:609–614
    [Google Scholar]
  36. Sundstrom P.M., Tam M.R., Nichols E.J., Kenny G.E. Antigenic differences in the surface mannoproteins of Candida albicans as revealed by monoclonal antibodies. Infect Immun 1988; 56:601–606
    [Google Scholar]
  37. Torosantucci A., Boccanera M., Casalinuovo I., Pellegrini G., Cassone A. Differences in the antigenic expression of immunomodulatory mannoprotein constituents on yeast and mycelial forms of Candida albicans. J Gen Microbiol 1990; 136:1421–1428
    [Google Scholar]
  38. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76:4350–4354
    [Google Scholar]
  39. Tsuchiya T., Fukazawa Y., Taguchi M., Nakase T., Shinoda T. Serologic aspects of yeast classification. Mycopathol My col Appi 1974; 53:77–91
    [Google Scholar]
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