Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1987 Mar;55(3):742–748. doi: 10.1128/iai.55.3.742-748.1987

Production, characterization, and antibody specificity of a mouse monoclonal antibody reactive with Cryptococcus neoformans capsular polysaccharide.

F Dromer, J Salamero, A Contrepois, C Carbon, P Yeni
PMCID: PMC260404  PMID: 3546139

Abstract

Two monoclonal immunoglobulin G1 antibodies reacting with Cryptococcus neoformans capsular polysaccharide (CNPS) were produced in mice by using a carefully defined procedure for immunization with unmodified CNPS purified from C. neoformans serotype A. Since the antibodies were found to have the same pattern of specificity, only one of them (E1) is described. This anti-CNPS monoclonal antibody reacted with the glucuronoxylomannan component of CNPS but not with the constituent monosaccharides or with the mannose alpha(1----3)-linked oligosaccharide structures present on CNPS. E1 appeared to be specific for C. neoformans serotype A by agglutination of whole cells; it was specific for soluble CNPS A by gel immunoprecipitation. However, indirect immunofluorescence and competitive-binding enzyme-linked immunosorbent assay experiments showed low levels of cross-reactivity with serotypes B and D but not with serotype C. Concentrations 10,000 times higher for serotypes B and D cells than for serotype A cells were required for a 50% inhibition of E1 anti-CNPS A activity as measured by enzyme-linked immunosorbent assay. Among the other yeasts tested, a cross-reaction was only detected with Trichosporon beigelii. The four serotypes of C. neoformans could be distinguished based on intensities and patterns of fluorescence in an indirect immunofluorescence assay using the monoclonal anti-CNPS A antibody. Monoclonal anti-CNPS A antibodies could be useful for fundamental studies on the glucuronoxylomannan structure, as well as for clinical applications such as serotyping and possibly the serological diagnosis of cryptococcosis.

Full text

PDF
742

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atkinson A. J., Jr, Bennett J. E. Experience with a new skin test antigen prepared from Cryptococcus neoformans. Am Rev Respir Dis. 1968 Apr;97(4):637–643. doi: 10.1164/arrd.1968.97.4.637. [DOI] [PubMed] [Google Scholar]
  2. BENNETT J. E., HASENCLEVER H. F. CRYPTOCOCCUS NEOFORMANS POLYSACCHARIDE: STUDIES OF SEROLOGIC PROPERTIES AND ROLE IN INFECTION. J Immunol. 1965 Jun;94:916–920. [PubMed] [Google Scholar]
  3. Bindschadler D. D., Bennett J. E. Serology of human cryptococcosis. Ann Intern Med. 1968 Jul;69(1):45–52. doi: 10.7326/0003-4819-69-1-45. [DOI] [PubMed] [Google Scholar]
  4. Biozzi G., Stiffel C., Mouton D., Liacopoulosbriot M., Decreusefond C., Bouthillier Y. Etude du phénomène de l'immuno-cyto-adhérence au cours de l'immunisation. Ann Inst Pasteur (Paris) 1966 Mar;110(3 Suppl):7–32. [PubMed] [Google Scholar]
  5. 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]
  6. Dromer F., Charreire J., Contrepois A., Carbon C., Yeni P. Protection of mice against experimental cryptococcosis by anti-Cryptococcus neoformans monoclonal antibody. Infect Immun. 1987 Mar;55(3):749–752. doi: 10.1128/iai.55.3.749-752.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Drutz D. J. Antigen detection in fungal infections. N Engl J Med. 1986 Jan 9;314(2):115–117. doi: 10.1056/NEJM198601093140210. [DOI] [PubMed] [Google Scholar]
  8. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 1971 Sep;8(9):871–874. doi: 10.1016/0019-2791(71)90454-x. [DOI] [PubMed] [Google Scholar]
  9. Fleet G. H. Composition and structure of yeast cell walls. Curr Top Med Mycol. 1985;1:24–56. doi: 10.1007/978-1-4613-9547-8_2. [DOI] [PubMed] [Google Scholar]
  10. Hall N. K., Blackstock R. Production of specific antibody to Cryptococcus neoformans by hybridomas in vitro. Sabouraudia. 1981 Jun;19(2):157–160. [PubMed] [Google Scholar]
  11. Henderson D. K., Bennett J. E., Huber M. A. Long-lasting, specific immunologic unresponsiveness associated with cryptococcal meningitis. J Clin Invest. 1982 May;69(5):1185–1190. doi: 10.1172/JCI110555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Kaplan W., Bragg S. L., Crane S., Ahearn D. G. Serotyping Cryptococcus neoformans by immunofluorescence. J Clin Microbiol. 1981 Sep;14(3):313–317. doi: 10.1128/jcm.14.3.313-317.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Kozel T. R., Cazin J., Jr Immune response to Cryptococcus neoformans soluble polysaccharide. I. Serological assay for antigen and antibody. Infect Immun. 1972 Jan;5(1):35–41. doi: 10.1128/iai.5.1.35-41.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kozel T. R., Hermerath C. A. Binding of cryptococcal polysaccharide to Cryptococcus neoformans. Infect Immun. 1984 Mar;43(3):879–886. doi: 10.1128/iai.43.3.879-886.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lewis J. L., Rabinovich S. The wide spectrum of cryptococcal infections. Am J Med. 1972 Sep;53(3):315–322. doi: 10.1016/0002-9343(72)90174-x. [DOI] [PubMed] [Google Scholar]
  18. McManus E. J., Jones J. M. Detection of a Trichosporon beigelii antigen cross-reactive with Cryptococcus neoformans capsular polysaccharide in serum from a patient with disseminated Trichosporon infection. J Clin Microbiol. 1985 May;21(5):681–685. doi: 10.1128/jcm.21.5.681-685.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Murphy J. W., Cozad G. C. Immunological unresponsiveness induced by cryptococcal capsular polysaccharide assayed by the hemolytic plaque technique. Infect Immun. 1972 Jun;5(6):896–901. doi: 10.1128/iai.5.6.896-901.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reiss E., Huppert M., Cherniak R. Characterization of protein and mannan polysaccharide antigens of yeasts, moulds, and actinomycetes. Curr Top Med Mycol. 1985;1:172–207. doi: 10.1007/978-1-4613-9547-8_7. [DOI] [PubMed] [Google Scholar]
  21. Sharon J., Kabat E. A., Morrison S. L. Studies on mouse hybridomas secreting IgM or IgA antibodies to alpha(1 to 6)-linked dextran. Mol Immunol. 1981 Sep;18(9):831–846. doi: 10.1016/0161-5890(81)90005-5. [DOI] [PubMed] [Google Scholar]
  22. Turner S. H., Cherniak R., Reiss E. Fractionation and characterization of galactoxylomannan from Cryptococcus neoformans. Carbohydr Res. 1984 Feb 15;125(2):343–349. doi: 10.1016/0008-6215(84)85172-1. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Wu T. C., Koo S. Y. Comparison of three commercial cryptococcal latex kits for detection of cryptococcal antigen. J Clin Microbiol. 1983 Nov;18(5):1127–1130. doi: 10.1128/jcm.18.5.1127-1130.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES