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. 1993 Jul;61(7):2879–2885. doi: 10.1128/iai.61.7.2879-2885.1993

Specificity of Cryptococcus neoformans factor sera determined by enzyme-linked immunosorbent assay and dot enzyme assay.

T Belay 1, R Cherniak 1, T Shinoda 1
PMCID: PMC280934  PMID: 7685739

Abstract

An indirect enzyme-linked immunosorbent assay (ELISA) and a dot enzyme assay (DEA) were used to determine the specificities of Cryptococcus neoformans factor sera to serotype type-specific capsular polysaccharides, glucuronoxylomannans (GXMs). Pure and chemically characterized GXMs were obtained from representative isolates of C. neoformans serotypes A, B, C, and D. Distinctive specificity patterns and quantitative differences were observed for each factor serum when the selected GXMs were studied by ELISA. The specificity patterns for each factor serum determined by DEA almost completely paralleled the ELISA results. The serotype specificities demonstrated by ELISA and DEA were similar to previously reported results that were obtained by slide agglutination studies of whole cells. On the basis of the ELISA and DEA activity patterns, factor sera 5, 6, and 8 were specific for serotypes B, C, and D, respectively; factor serum 1 was strongly reactive to all serotypes; factor serum 2 was specific for serotypes A, B, and D; factor serum 3 was specific for serotypes A and D; and factor serum 4 was specific for serotypes B and C. The specificity of factor serum 7 for serotype A was demonstrated by DEA only. Structural variation was indicated among the serotype C isolates studied because a unique activity pattern versus factor serum 6 was observed for each isolate. The quantitative differences in the activity of the GXMs from five serotype C isolates suggest that mannopyranoside residues substituted O-2 and O-4 with xylose are essential elements of the determinant responsible for the observed activity of factor 6. No significant differences in activity patterns and specificities of factor serum 6 were observed when O-deacetylated GXMs were substituted for the native GXMs. Our results show that ELISA and DEA are valuable techniques for the serological analysis of cryptococcal factor sera and GXMs.

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Selected References

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  1. Bhattacharjee A. K., Bennett J. E., Glaudemans C. P. Capsular polysaccharides of Cryptococcus neoformans. Rev Infect Dis. 1984 Sep-Oct;6(5):619–624. doi: 10.1093/clinids/6.5.619. [DOI] [PubMed] [Google Scholar]
  2. Bulmer G. S., Sans M. D. Cryptococcus neoformans. 3. Inhibition of phagocytosis. J Bacteriol. 1968 Jan;95(1):5–8. doi: 10.1128/jb.95.1.5-8.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Casadevall A., Mukherjee J., Devi S. J., Schneerson R., Robbins J. B., Scharff M. D. Antibodies elicited by a Cryptococcus neoformans-tetanus toxoid conjugate vaccine have the same specificity as those elicited in infection. J Infect Dis. 1992 Jun;165(6):1086–1093. doi: 10.1093/infdis/165.6.1086. [DOI] [PubMed] [Google Scholar]
  4. Casadevall A., Mukherjee J., Scharff M. D. Monoclonal antibody based ELISAs for cryptococcal polysaccharide. J Immunol Methods. 1992 Sep 18;154(1):27–35. doi: 10.1016/0022-1759(92)90209-c. [DOI] [PubMed] [Google Scholar]
  5. Cherniak R., Cheeseman M. M., Reyes G. H., Reiss E., Todaro F. Enhanced binding of capsular polysaccharides of Cryptococcus neoformans to polystyrene microtitration plates for enzyme-linked immunosorbent assay. Diagn Clin Immunol. 1988;5(6):344–348. [PubMed] [Google Scholar]
  6. Cherniak R., Morris L. C., Anderson B. C., Meyer S. A. Facilitated isolation, purification, and analysis of glucuronoxylomannan of Cryptococcus neoformans. Infect Immun. 1991 Jan;59(1):59–64. doi: 10.1128/iai.59.1.59-64.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cherniak R., Morris L. C., Meyer S. A. Glucuronoxylomannan of Cryptococcus neoformans serotype C: structural analysis by gas-liquid chromatography-mass spectrometry and 13C-nuclear magnetic resonance spectroscopy. Carbohydr Res. 1992 Mar 2;225(2):331–337. doi: 10.1016/s0008-6215(00)90505-6. [DOI] [PubMed] [Google Scholar]
  8. Cherniak R., Reiss E., Slodki M. E., Plattner R. D., Blumer S. O. Structure and antigenic activity of the capsular polysaccharide of Cryptococcus neoformans serotype A. Mol Immunol. 1980 Aug;17(8):1025–1032. doi: 10.1016/0161-5890(80)90096-6. [DOI] [PubMed] [Google Scholar]
  9. Dromer F., Salamero J., Contrepois A., Carbon C., Yeni P. Production, characterization, and antibody specificity of a mouse monoclonal antibody reactive with Cryptococcus neoformans capsular polysaccharide. Infect Immun. 1987 Mar;55(3):742–748. doi: 10.1128/iai.55.3.742-748.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. EVANS E. E. The antigenic composition of Cryptococcus neoformans. I. A serologic classification by means of the capsular and agglutination reactions. J Immunol. 1950 May;64(5):423–430. [PubMed] [Google Scholar]
  11. Eckert T. F., Kozel T. R. Production and characterization of monoclonal antibodies specific for Cryptococcus neoformans capsular polysaccharide. Infect Immun. 1987 Aug;55(8):1895–1899. doi: 10.1128/iai.55.8.1895-1899.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ikeda R., Nishikawa A., Shinoda T., Fukazawa Y. Chemical characterization of capsular polysaccharide from Cryptococcus neoformans serotype A-D. Microbiol Immunol. 1985;29(10):981–991. doi: 10.1111/j.1348-0421.1985.tb02962.x. [DOI] [PubMed] [Google Scholar]
  13. Ikeda R., Shinoda T., Fukazawa Y., Kaufman L. Antigenic characterization of Cryptococcus neoformans serotypes and its application to serotyping of clinical isolates. J Clin Microbiol. 1982 Jul;16(1):22–29. doi: 10.1128/jcm.16.1.22-29.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kaplan M. H., Rosen P. P., Armstrong D. Cryptococcosis in a cancer hospital: clinical and pathological correlates in forty-six patients. Cancer. 1977 May;39(5):2265–2274. doi: 10.1002/1097-0142(197705)39:5<2265::aid-cncr2820390546>3.0.co;2-x. [DOI] [PubMed] [Google Scholar]
  15. Kovacs J. A., Kovacs A. A., Polis M., Wright W. C., Gill V. J., Tuazon C. U., Gelmann E. P., Lane H. C., Longfield R., Overturf G. Cryptococcosis in the acquired immunodeficiency syndrome. Ann Intern Med. 1985 Oct;103(4):533–538. doi: 10.7326/0003-4819-103-4-533. [DOI] [PubMed] [Google Scholar]
  16. Kwon-Chung K. J., Bennett J. E. Epidemiologic differences between the two varieties of Cryptococcus neoformans. Am J Epidemiol. 1984 Jul;120(1):123–130. doi: 10.1093/oxfordjournals.aje.a113861. [DOI] [PubMed] [Google Scholar]
  17. Pettoello-Mantovani M., Casadevall A., Kollmann T. R., Rubinstein A., Goldstein H. Enhancement of HIV-1 infection by the capsular polysaccharide of Cryptococcus neoformans. Lancet. 1992 Jan 4;339(8784):21–23. doi: 10.1016/0140-6736(92)90142-p. [DOI] [PubMed] [Google Scholar]
  18. Scott E. N., Muchmore H. G., Felton F. G. Enzyme-linked immunosorbent assays in murine cryptococcosis. Sabouraudia. 1981 Dec;19(4):257–265. doi: 10.1080/00362178185380431. [DOI] [PubMed] [Google Scholar]
  19. Spiropulu C., Eppard R. A., Otteson E., Kozel T. R. Antigenic variation within serotypes of Cryptococcus neoformans detected by monoclonal antibodies specific for the capsular polysaccharide. Infect Immun. 1989 Oct;57(10):3240–3242. doi: 10.1128/iai.57.10.3240-3242.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Todaro-Luck F., Reiss E., Cherniak R., Kaufman L. Characterization of Cryptococcus neoformans capsular glucuronoxylomannan polysaccharide with monoclonal antibodies. Infect Immun. 1989 Dec;57(12):3882–3887. doi: 10.1128/iai.57.12.3882-3887.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Turner S. H., Cherniak R. Glucuronoxylomannan of Cryptococcus neoformans serotype B: structural analysis by gas-liquid chromatography-mass spectrometry and 13C-nuclear magnetic resonance spectroscopy. Carbohydr Res. 1991 Apr 2;211(1):103–116. doi: 10.1016/0008-6215(91)84149-9. [DOI] [PubMed] [Google Scholar]
  22. Wilson D. E., Bennett J. E., Bailey J. W. Serologic grouping of Cryptococcus neoformans. Proc Soc Exp Biol Med. 1968 Mar;127(3):820–823. doi: 10.3181/00379727-127-32812. [DOI] [PubMed] [Google Scholar]
  23. Zuger A., Louie E., Holzman R. S., Simberkoff M. S., Rahal J. J. Cryptococcal disease in patients with the acquired immunodeficiency syndrome. Diagnostic features and outcome of treatment. Ann Intern Med. 1986 Feb;104(2):234–240. doi: 10.7326/0003-4819-104-2-234. [DOI] [PubMed] [Google Scholar]

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