1887

Abstract

Different exocellular extracts were isolated by concentrating the supernatants of yeast- and mycelial-phase cultures incubated in a synthetic medium. The only difference between the extracts obtained from the two phases was the presence in those obtained from mycelial cultures of a polysaccharide-rich, high-molecular-mass component, migrating in SDS-polyacrylamide gels at a position that would correspond to proteins with molecular masses of 245–265 kDa. The electrophoretic band patterns obtained before and after concanavalin A-Sepharose 4B affinity column treatments confirmed that the 245–265 kDa band was the only one of mannoprotein nature. The extract obtained from 24 h mycelial-phase culture (EA) was selected as the exocellular antigen for this work. The dry weight of EA obtained from 1 litre of culture medium was 30 mg; it contained 53% carbohydrate (18·3% glucose and 21·7% mannose measured by gas-liquid chromatography) and 10% protein. Rabbit antisera against EA were absorbed with yeast-phase organisms and used to stain Western blots of gels loaded with EAs. These antisera clearly recognized bands in the 21, 33 and 44 kDa areas. The antiserum obtained was employed to develop a double-antibody enzyme-linked immunosorbent assay for measuring EA concentrations in a culture medium. Most of the EA was released during the exponential phase of growth.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-12-3005
1993-12-01
2024-12-03
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/12/mic-139-12-3005.html?itemId=/content/journal/micro/10.1099/00221287-139-12-3005&mimeType=html&fmt=ahah

References

  1. Bailey J.W., Sada E., Brass E., Bennett J.E. 1985; Diagnosis of systemic candidiasis by latex agglutination for serum antigen. Journal of Clinical Microbiology 21:749–752
    [Google Scholar]
  2. Casanova M., Gil M.L., Cardeñoso L., Martinez J.P., Sentandreu R. 1989; Identification of wall-specific antigens synthesized during germ tube formation by Candida albicans. Infection and Immunity 57:262–271
    [Google Scholar]
  3. Elorza M.V., Rico H., Gozalbo D., Sentandreu R. 1983; Cell wall composition and protoplast regeneration in Candida albicans. Antonie van Leeuwenhoek 49:457–469
    [Google Scholar]
  4. Kapitany R.A., Zebrowsky E.J. 1973; A high resolution PAS stain for polyacrylamide gel electrophoresis. Analytical Biochemistry 56:361–369
    [Google Scholar]
  5. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  6. Lee J.C., King R.D. 1983; Characterization of Candida albicans adherence to human vaginal epithelial cells in vitro. Infection and Immunity 41:1024–1030
    [Google Scholar]
  7. Lee K.L., Buckley H.R., Campbell C.C. 1975; An amino acid liquid synthetic medium for the development of mycelial and yeast forms of Candida albicans. Sabouraudia 13:148–153
    [Google Scholar]
  8. Loewus F.A. 1952; Improvement in the anthrone method for determination of carbohydrates. Analytical Chemistry 24:219–222
    [Google Scholar]
  9. Louria D.B., Brayton R.G., Finkel G. 1963; Studies on the pathogenesis of experimental Candida albicans infections in mice. Sabouraudia 2:271–283
    [Google Scholar]
  10. Macdonald F., Odds F.C. 1980; Purified Candida albicans proteinase in the serological diagnosis of systemic candidosis. Journal of the American Medical Association 243:2409–2411
    [Google Scholar]
  11. Mankowski Z.T. 1968; Production of glycoprotein by Candida albicans in a synthetic medium and its influence on the growth of newborn mice. Mycopathologia et Mycologia Applicata 36:113–118
    [Google Scholar]
  12. Manning-Zweerink M., Maloney C.S., Mitchell T.G., Weston H. 1986; Immunoblot analyses of Candida albicans- associated antigens and antibodies in human sera. Journal of Clinical Microbiology 23:46–52
    [Google Scholar]
  13. Masler L., Sikl D., Bauer S., Sandula J. 1966; Extracellular polysaccharide-protein complexes produced by selected strains of Candida albicans. Folia Microbiologica 11:373–378
    [Google Scholar]
  14. Mccourtie J., Douglas L.J. 1985; Extracellular polymer of Candida albicans: isolation, analysis and role in adhesion. Journal of General Microbiology 131:495–503
    [Google Scholar]
  15. Meckstroth K.L., Reiss E., Keller J.W., Kaufman L. 1981; Detection of antibodies and antigenemia in patients with candidiasis by enzyme-linked immunosorbent assay. Journal of Infectious Diseases 144:24–32
    [Google Scholar]
  16. Mitchell A.J., Scurfield G. 1970; An assessment of infrared spectra as indicators of fungal cell wall composition. Australian Journal of Biological Sciences 23:345–360
    [Google Scholar]
  17. Nakene P.K., Kawoi A. 1974; Peroxidase-labelled antibody, a new method of conjugation. Journal of Histochemistry and Cytochemistry 22:1084–1091
    [Google Scholar]
  18. Odds F.C. 1988; Introduction and historical note. In Candida and Candidosis, 2nd edn. pp. 1–5 Philadelphia: Bailliere Tindall;
    [Google Scholar]
  19. Ponton J., Jones J.M. 1986; Identification of two germ-tube- specific cell wall antigens of Candida albicans. Infection and Immunity 54:864–868
    [Google Scholar]
  20. Scherwitz C. 1982; Ultrastructure of human cutaneous candidiasis. Journal of Investigative Dermatology 78:200–205
    [Google Scholar]
  21. Sikl D., Masler L., Bauer S. 1964; Mannan from the extracellular surface of Candida albicans Berkhourt. Experientia 20:456
    [Google Scholar]
  22. Smail E.H., Jones J.M. 1984; Demonstration and solubilization of antigens expressed primarily on the surfaces of Candida albicans germ tubes. Infection and Immunity 45:74–81
    [Google Scholar]
  23. Smith C.B. 1985; Candidiasis: pathogenesis, host resistance and predisposing factors. In Candidiasis pp. 53–70 Bodey G. P., Fainstein V. Edited by New York: Raven Press;
    [Google Scholar]
  24. Sobel J.D., Muller G., Buckley H.R. 1984; Critical role of germ tube formation in the pathogenesis of candidal vaginitis. Infection and Immunity 44:576–580
    [Google Scholar]
  25. Sundstrom P.M., Kenny G.E. 1984; Characterization of antigens specific to the surface of germ tubes of Candida albicans by immunofluorescence. Infection and Immunity 43:850–855
    [Google Scholar]
  26. Sundstrom P.M., Kenny G.E. 1985; Enzymatic release of germtube-specific antigens from cell walls of Candida albicans. Infection and Immunity 49:609–614
    [Google Scholar]
  27. Torosantucci A., Gomez M.J., Casalinuovo I., Cassone A. 1991; Biochemical and antigenic characterization of mannoprotein constituents released from yeast and mycelial forms of Candida albicans. Journal of Medical and Veterinary Mycology 29:361–372
    [Google Scholar]
  28. Tsachdjian C.L., Kozinn P.J., Okas A., Caroline L., Halle A. 1967; Serodiagnosis of systemic candidiasis. Journal of Infectious Diseases 117:180–187
    [Google Scholar]
  29. Vaitukaitis J.L. 1981; Production of antisera with small doses of immunogen: multiple intradermal injections. Methods in Enzymology 73:46–51
    [Google Scholar]
/content/journal/micro/10.1099/00221287-139-12-3005
Loading
/content/journal/micro/10.1099/00221287-139-12-3005
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error