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. 1983 Jun;71(6):1602–1613. doi: 10.1172/JCI110916

Specific in vitro antimannan-rich antigen of Candida albicans antibody production by sensitized human blood lymphocytes.

A Durandy, A Fischer, C Griscelli
PMCID: PMC370366  PMID: 6345586

Abstract

We have developed a new antigenic system for the induction of specific in vitro antibody response in man. The antigen used was purified from the cell wall of Candida albicans strain A and contained greater than 96% polysaccharide mannan. Peripheral blood mononuclear cells from Candida-sensitized donors produced specific antimannan antibodies during a 7-d culture in the presence of mannan absorbed with methylated bovine serum albumin. Two methods were used to detect antimannan antibody responses. Antimannan antibody-producing cells were identified by radioautography with tritiated mannan. Antibody concentration in culture supernatants was measured by an enzyme-linked immunosorbent assay. In both methods, specific IgM and IgG (but not IgA) antibodies were detected. The antibody production to mannan was specific, since an antigenically unrelated polysaccharide (pneumococcal antigen S III) did not bind to methylated bovine serum albumin-mannan-induced blast cells and did not induce antimannan antibody-containing cells. Furthermore, a pulse with an excess of unlabeled mannan abolished [3H]mannan binding, whereas an excess of unlabeled S III did not. Similarly, no antimannan antibody was obtained in influenza virus-stimulated cultures and mannan-stimulated cultures were not inducing antiinfluenza antibodies. The antimannan antibody production was shown to be a T cell-dependent phenomenon. The T helper effect appeared to be radiosensitive. It was under a genetic restriction as it occurred only in autologous or semi-identical but not in allogeneic situations. This system is relatively simple, reproducible, and well suited for the study of specific secondary in vitro antibody responses to polysaccharide antigens in humans.

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Selected References

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  1. Axelsen N. H. Analysis of human Candida precipitins by quantitative immunoelectrophoresis: a model for analysis of complex microbial antigen-antibody systems. Scand J Immunol. 1976;5(3):177–190. doi: 10.1111/j.1365-3083.1976.tb00268.x. [DOI] [PubMed] [Google Scholar]
  2. Axelsen N. H., Kirkpatrick C. H. Simultaneous characterization of free Candida antigens and Candida precipitins in a patient's serum by means of crossed immunoelectrophoresis with intermidiate gel. J Immunol Methods. 1973 Apr;2(3):245–249. doi: 10.1016/0022-1759(73)90050-1. [DOI] [PubMed] [Google Scholar]
  3. Baker P. H., Stashak P. W. Quantitative and qualitative studies on the primary antibody response to pneumococcal polysaccharides at ehe cellular level. J Immunol. 1969 Dec;103(6):1342–1348. [PubMed] [Google Scholar]
  4. Banck G., Forsgren A. Many bacterial species are mitogenic for human blood B lymphocytes. Scand J Immunol. 1978;8(4):347–354. doi: 10.1111/j.1365-3083.1978.tb00528.x. [DOI] [PubMed] [Google Scholar]
  5. Blaese R. M., Strober W., Brown R. S., Waldmann T. A. The Wiskott-Aldrich syndrome. A disorder with a possible defect in antigen processing or recognition. Lancet. 1968 May 18;1(7551):1056–1061. doi: 10.1016/s0140-6736(68)91411-6. [DOI] [PubMed] [Google Scholar]
  6. Bona C., Broder S., Dimitriu A., Waldmann T. A. Polyclonal activation of human B lymphocytes by Nocardia water soluble mitogen (NWSM). Immunol Rev. 1979;45:69–92. doi: 10.1111/j.1600-065x.1979.tb00273.x. [DOI] [PubMed] [Google Scholar]
  7. Callard R. E., Smith C. M. Histocompatibility requirements for T cell help in specific in vitro antibody responses to influenza virus by human blood lymphocytes. Eur J Immunol. 1981 Mar;11(3):206–212. doi: 10.1002/eji.1830110309. [DOI] [PubMed] [Google Scholar]
  8. Callard R. E. Specific in vitro antibody response to influenza virus by human blood lymphocytes. Nature. 1979 Dec 13;282(5740):734–736. doi: 10.1038/282734a0. [DOI] [PubMed] [Google Scholar]
  9. Delfraissy J. F., Galanaud P., Dormont J., Wallon C. Primary in vitro antibody response from human peripheral blood lymphocytes. J Immunol. 1977 Feb;118(2):630–635. [PubMed] [Google Scholar]
  10. Dosch H-M, Gelfand E. W. Generation of human plaque-forming cells in culture: tissue distribution, antigenic and cellular requirements. J Immunol. 1977 Jan;118(1):302–308. [PubMed] [Google Scholar]
  11. Durandy A., Fischer A., Griscelli C. Active suppression of B lymphocyte maturation by two different newborn T lymphocyte subsets. J Immunol. 1979 Dec;123(6):2644–2650. [PubMed] [Google Scholar]
  12. 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]
  13. FELTON L. D., PRESCOTT B., KAUFFMANN G., OTTINGER B. Pneumococcal antigenic polysaccharide substances from animal tissues. J Immunol. 1955 Mar;74(3):205–213. [PubMed] [Google Scholar]
  14. Fauci A. S., Pratt K. R. Activation of human B lymphocytes. I. Direct plaque-forming cell assay for the measurement of polyclonal activation and antigenic stimulation of human B lymphocytes. J Exp Med. 1976 Sep 1;144(3):674–684. doi: 10.1084/jem.144.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Geha R. S., Schneeberger E., Rosen F. S., Merler E. Interaction of human thymus-derived and non-thymus-derived lymphocytes in vitro. Induction of proliferation and antibody synthesis in B lymphocytes by a soluble factor released from antigen-stimulated T lymphocytes. J Exp Med. 1973 Nov 1;138(5):1230–1247. doi: 10.1084/jem.138.5.1230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Heijnen C. J., Uytdehaag F., Pot K. H., Ballieux R. E. Antigen-specific human T cell factors. I. T cell helper factor: biololgic properties. J Immunol. 1981 Feb;126(2):497–502. [PubMed] [Google Scholar]
  18. Hoffmann M. K. Antigen-specific induction and regulation of antibody synthesis in cultures of human peripheral blood mononuclear cells. Proc Natl Acad Sci U S A. 1980 Feb;77(2):1139–1143. doi: 10.1073/pnas.77.2.1139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Klaus G. G., Humphrey J. H. Mechanisms of b cell triggering: studies with t cell-independent antigens. Transplant Rev. 1975;23:105–118. doi: 10.1111/j.1600-065x.1975.tb00152.x. [DOI] [PubMed] [Google Scholar]
  20. Kuritani T., Cooper M. D. Human B cell differentiation. II. Pokeweed mitogen-responsive B cells belong to a surface immunoglobulin D-negative subpopulation. J Exp Med. 1982 May 1;155(5):1561–1566. doi: 10.1084/jem.155.5.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lane H. C., Volkman D. J., Whalen G., Fauci A. S. In vitro antigen-induced, antigen-specific antibody production in man. Specific and polyclonal components, kinetics, and cellular requirements. J Exp Med. 1981 Oct 1;154(4):1043–1057. doi: 10.1084/jem.154.4.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miller R. A., Gartner S., Kaplan H. S. Stimulation of mitogenic responses in human peripheral blood lymphocytes by lipopolysaccharide: serum and T helper cell requirements. J Immunol. 1978 Dec;121(6):2160–2164. [PubMed] [Google Scholar]
  23. Mongini P. K., Paul W. E., Metcalf E. S. T cell regulation of immunoglobulin class expression in the antibody response to trinitrophenyl-ficoll. Evidence for T cell enhancement of the immunoglobulin class switch. J Exp Med. 1982 Mar 1;155(3):884–902. doi: 10.1084/jem.155.3.884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Phan-Dinh-Tuy F., Durandy A., Griscelli C., Bach M. A. T-cell subset analysis by monoclonal antibodies in primary immunodeficiencies. Scand J Immunol. 1981 Aug;14(2):193–200. doi: 10.1111/j.1365-3083.1981.tb00199.x. [DOI] [PubMed] [Google Scholar]
  25. Ringdén O., Rynnel-Dagö B., Waterfield E. M., Möller E., Möller G. Polyclonal antibody secretion in human lymphocytes induced by killed staphylococcal bacteria and by lipopolysaccharide. Scand J Immunol. 1977;6(11):1159–1169. doi: 10.1111/j.1365-3083.1977.tb00355.x. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Sager D. S., Jasin H. E. Bacterial lipopolysaccharide-induced immunoglobulin synthesis by human blood lymphocytes partially depleted of monocytes. Clin Exp Immunol. 1982 Mar;47(3):645–652. [PMC free article] [PubMed] [Google Scholar]
  28. Siskind G. W., Benacerraf B. Cell selection by antigen in the immune response. Adv Immunol. 1969;10:1–50. doi: 10.1016/s0065-2776(08)60414-9. [DOI] [PubMed] [Google Scholar]
  29. Thomas Y., Rogozinski L., Irigoyen O. H., Shen H. H., Talle M. A., Goldstein G., Chess L. Functional analysis of human T cell subsets defined by monoclonal antibodies. V. Suppressor cells within the activated OKT4+ population belong to a distinct subset. J Immunol. 1982 Mar;128(3):1386–1390. [PubMed] [Google Scholar]
  30. Thomas Y., Sosman J., Irigoyen O., Friedman S. M., Kung P. C., Goldstein G., Chess L. Functional analysis of human T cell subsets defined by monoclonal antibodies. I. Collaborative T-T interactions in the immunoregulation of B cell differentiation. J Immunol. 1980 Dec;125(6):2402–2408. [PubMed] [Google Scholar]
  31. Volkman D. J., Lane H. C., Fauci A. S. Antigen-induced in vitro antibody production in humans: a model for B cell activation and immunoregulation. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2528–2531. doi: 10.1073/pnas.78.4.2528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wiener E., Bandieri A. Differences in antigen handling by peritoneal macrophages from the Biozzi high and low responder lines of mice. Eur J Immunol. 1974 Jul;4(7):457–463. doi: 10.1002/eji.1830040703. [DOI] [PubMed] [Google Scholar]
  33. Yarchoan R., Murphy B. R., Strober W., Schneider H. S., Nelson D. L. Specific anti-influenza virus antibody production in vitro by human peripheral blood mononuclear cells. J Immunol. 1981 Dec;127(6):2588–2594. [PubMed] [Google Scholar]

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