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. 1986 Nov;24(5):796–802. doi: 10.1128/jcm.24.5.796-802.1986

Monoclonal antibodies against Candida tropicalis mannan: antigen detection by enzyme immunoassay and immunofluorescence.

E Reiss, L de Repentigny, R J Kuykendall, A W Carter, R Galindo, P Auger, S L Bragg, L Kaufman
PMCID: PMC269031  PMID: 2429989

Abstract

Three strains of mice were immunized with Candida tropicalis cell walls, and antibodies against mannan were detected by indirect enzyme immunoassay (EIA) in 3 of 9 BALB/c mice, 4 of 11 C57BL/6 mice, and 4 of 8 CFW mice. Responding mice produced immunoglobulin M (IgM), but IgG was not detected in their sera. Fusion of the high-responder BALB/c mouse with a plasmacytoma cell line resulted in 41 clones secreting antimannan monoclonal antibodies (MAbs). Four clones selected for propagation included one IgM and one IgG MAb that reacted with mannans of Candida albicans serotypes A and B and of C. tropicalis and two IgM MAbs specific for an epitope only in the mannans of C. albicans serotype A and C. tropicalis. One of the IgM MAbs, CB6, was an effective substitute for rabbit antibodies in the double-antibody sandwich EIA to detect antigenemia produced in rabbits infected with C. albicans A or C. tropicalis. It could function either as the peroxidase-conjugated indicator antibody or as the capture antibody. Two MAbs, CB6 (C. tropicalis and C. albicans A specific) and AC3 (C. tropicalis and C. albicans A and B specific), functioned in place of polyclonal antisera in the serotyping of C. albicans by immunofluorescence. There was 95.8% agreement in the results of serotyping using MAbs as reagents compared with rabbit antisera. Competitive inhibition in EIA between CB6 and monospecific antisera against C. albicans factors 1, 4, and 6 indicated that CB6 binds to an epitope which is probably factor 6. Serologic similarity between factor 4 and the binding site of MAb AC3 was also determined.

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Selected References

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  1. Auger P., Dumas C., Joly J. A study of 666 strains of Candida albicans: correlation between serotype and susceptibility to 5-fluorocytosine. J Infect Dis. 1979 May;139(5):590–594. doi: 10.1093/infdis/139.5.590. [DOI] [PubMed] [Google Scholar]
  2. Ballou C. Structure and biosynthesis of the mannan component of the yeast cell envelope. Adv Microb Physiol. 1976;14(11):93–158. doi: 10.1016/s0065-2911(08)60227-1. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Fischer A., Ballet J. J., Griscelli C. Specific inhibition of in vitro Candida-induced lymphocyte proliferation by polysaccharidic antigens present in the serum of patients with chronic mucocutaneous candidiasis. J Clin Invest. 1978 Nov;62(5):1005–1013. doi: 10.1172/JCI109204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Lehmann P. F., Reiss E. Comparison by ELISA of serum anti-Candida albicans mannan IgG levels of a normal population and in diseased patients. Mycopathologia. 1980 Mar 17;70(2):89–93. doi: 10.1007/BF00443073. [DOI] [PubMed] [Google Scholar]
  6. Lehmann P. F., Reiss E. Detection of Candida albicans mannan by immunodiffusion, counterimmunoelectrophoresis, and enzyme-linked immunoassay. Mycopathologia. 1980 Mar 17;70(2):83–88. doi: 10.1007/BF00443072. [DOI] [PubMed] [Google Scholar]
  7. McDougal J. S., Browning S. W., Kennedy S., Moore D. D. Immunodot assay for determining the isotype and light chain type of murine monoclonal antibodies in unconcentrated hybridoma culture supernates. J Immunol Methods. 1983 Oct 28;63(3):281–290. doi: 10.1016/s0022-1759(83)80001-5. [DOI] [PubMed] [Google Scholar]
  8. Miyakawa Y., Kagaya K., Fukazawa Y., Soe G. Production and characterization of agglutinating monoclonal antibodies against predominant antigenic factors for Candida albicans. J Clin Microbiol. 1986 May;23(5):881–886. doi: 10.1128/jcm.23.5.881-886.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nakajima T., Ballou C. E. Characterization of the carbohydrate fragments obtained from Saccharomyces cerevisiae mannan by alkaline degradation. J Biol Chem. 1974 Dec 10;249(23):7679–7684. [PubMed] [Google Scholar]
  10. Okubo Y., Honma Y., Suzuki S. Relationship between phosphate content and serological activities of the mannans of Candida albicans strains NIH A-207, NIH B-792, and J-1012. J Bacteriol. 1979 Jan;137(1):677–680. doi: 10.1128/jb.137.1.677-680.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Piccolella E., Lombardi G., Morelli R. Human lymphocyte-activating properties of a purified polysaccharide from Candida albicans: B and T cell cooperation in the mitogenic response. J Immunol. 1980 Nov;125(5):2082–2088. [PubMed] [Google Scholar]
  12. Reiss E., Kuykendall R. J., Kaufman L. Antigenemia in rabbits infected with Candida albicans serotype B: detection by enzyme immunoassay and preliminary characterization of the antigen. J Med Vet Mycol. 1986 Aug;24(4):259–269. [PubMed] [Google Scholar]
  13. Reiss E., Patterson D. G., Yert L. W., Holler J. S., Ibrahim B. K. Structural analysis of mannans from Candida albicans serotypes A and B and from Torulopsis glabrata by methylation gas chromatography mass spectrometry and exo-alpha-mannanase. Biomed Mass Spectrom. 1981 Jun;8(6):252–255. doi: 10.1002/bms.1200080605. [DOI] [PubMed] [Google Scholar]
  14. Reiss E., Stone S. H., Hasenclever H. F. Serological and cellular immune activity of peptidoglucomannan fractions of Candida albicans cell walls. Infect Immun. 1974 May;9(5):881–890. doi: 10.1128/iai.9.5.881-890.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SUMMERS D. F., GROLLMAN A. P., HASENCLEVER H. F. POLYSACCHARIDE ANTIGENS OF CANDIDA CELL WALL. J Immunol. 1964 Mar;92:491–499. [PubMed] [Google Scholar]
  16. Shinoda T., Kaufman L., Padhye A. A. Comparative evaluation of the Iatron serological Candida check kit and the API 20C kit for identification of medically important Candida species. J Clin Microbiol. 1981 Mar;13(3):513–518. doi: 10.1128/jcm.13.3.513-518.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Suzuki M., Fukazawa Y. Immunochemical characterization of Candida albicans cell wall antigens: specific determinant of Candida albicans serotype A mannan. Microbiol Immunol. 1982;26(5):387–402. doi: 10.1111/j.1348-0421.1982.tb00189.x. [DOI] [PubMed] [Google Scholar]
  18. Tsuchiya T., Fukazawa Y., Taguchi M., Nakase T., Shinoda T. Serologic aspects on yeast classification. Mycopathol Mycol Appl. 1974 Aug 30;53(1):77–91. doi: 10.1007/BF02127199. [DOI] [PubMed] [Google Scholar]
  19. Zhang W. J., Ballou C. E. Saccharomyces kluyveri cell wall mannoprotein. Structures of the O- and N-linked carbohydrate components. J Biol Chem. 1981 Oct 10;256(19):10073–10079. [PubMed] [Google Scholar]
  20. de Repentigny L., Kuykendall R. J., Chandler F. W., Broderson J. R., Reiss E. Comparison of serum mannan, arabinitol, and mannose in experimental disseminated candidiasis. J Clin Microbiol. 1984 Jun;19(6):804–812. doi: 10.1128/jcm.19.6.804-812.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. de Repentigny L., Reiss E. Current trends in immunodiagnosis of candidiasis and aspergillosis. Rev Infect Dis. 1984 May-Jun;6(3):301–312. doi: 10.1093/clinids/6.3.301. [DOI] [PubMed] [Google Scholar]

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