Immunochemical Analysis of Histoplasmin Proteins and Polysaccharide

  • Errol Reiss
  • Sandra L. Bragg


Histoplasmin is the supernatant fluid obtained after static cultivation of the mycelial form of Histoplasma capsulatum for 3 to 6 months at ambient temperature (22 to 25°C) on asparagine-glucose-glycerol-salts medium (Smith et al, 1948). Three types of histoplasmin exist — skin test reagents from the mycelial and yeast forms and serological histoplasmin. The method of preparing skin test histoplasmin was established long ago (Shaw et al, 1950) and single lots were used widely for many years. The skin test antigen was standardized by its ability to elicit a reproducible cutaneous hypersensitivity response in sensitized humans. The antigens in histoplasmin responsible for the skin test have been inferred but no direct structure-activity evidence exists, because monomolecular antigens have not been available. Recently, histoplasmin from yeast form cultures has been approved for human use as a skin test antigen (Scalarone et al, 1985). The bulk of accumulated information concerns mycelial histoplasmin and there are far fewer data about the characteristics of the yeast form product. For that reason, and because of space limitation, the following discussion applies primarily to mycelial histoplasmin.


Skin Test Amino Sugar Yeast Form Mycelial Form Immunochemical Analysis 
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  1. Azuma, I, Kanetsuna, F., Tanaka, Y., Yamamura, Y., and Carbonell, L. M., 1974, Chemical and immunological properties of galactomannans obtained from Histoplasma duboisii, Histoplasma capsulatum, Paracoccidioides brasiliensis, and Blastomyces dermatitidis, Mycopathol. Mycol. Appl. 54: 111–125.PubMedCrossRefGoogle Scholar
  2. Brock, E. G., Reiss, E., Pine, L., and Kaufman, L., 1984, Effect of periodate oxidation on the detection of antibodies against the M-antigen of histoplasmin by enzyme immunoassay (EIA) — inhibition. Current Microbiol. 10: 177–180.CrossRefGoogle Scholar
  3. Campbell, C. C. and Hill, G. B., 1964, Further studies on the development of complement-fixing antibodies and precipitins in healthy histoplasmin-sensitive persons following a single histoplasmin skin test. Amer. Rev. Resp. Dis. 90: 927–934.PubMedGoogle Scholar
  4. Davis, T. E. Jr., Domer, J.E., and Li, Y. T., 1977, Cell wall studies of Histoplasma capsulatum and Blastomyces dermatitidis using autologous and heterologous enzymes, Infect. Immun. 15: 978–987.PubMedGoogle Scholar
  5. Department of Health and Human Services, U. S., 1984, “Biosafety in Microbiological and Biomedical Laboratories,” J. H. Richardson, W. E. Barkley, eds., U. S. Government Printing Office, Washington, D.C.Google Scholar
  6. Dickerson, Q. H. Jr., and Busey, J. F., 1968, Chromatographic separation of h and m antigens from histoplasmin, Proc. Soc. Exper. Biol. Med. 128: 654–658.Google Scholar
  7. Ehrhard, H.-B. and Pine, L., 1972, Factors influencing the production of H and M antigens by Histoplasma capsulatum: Development and evaluation of a shake culture procedure, Appl. Microbiol. 23: 236–249.PubMedGoogle Scholar
  8. Ellsworth, J. H., Reiss, E., Bradley, R. L., Chmel, H., and Armstrong, D., 1977, Comparative serological and cutaneous reactivity of candidal cytoplasmic proteins and mannan separated by affinity for concanavalin A, J. Clin. Microbiol. 5: 91–99.PubMedGoogle Scholar
  9. Food and Drug Administration, U. S., 1977, Skin test antigens, proposed implementation of efficacy review. Fed. Register, 42, 52674–52687.Google Scholar
  10. Goodwin, R. A. Jr., and Des-Prez, R. M., 1978, State of the art — histoplasmosis, Amer. Rev. Resp. Dis. 117: 929–955.PubMedGoogle Scholar
  11. Green, J. H., Harrell, W. K., Gray, S. B., Johnson, J. E., Bolin, R. C., Gross, H. and Malcolm, G. B., 1976, H and M antigens of Histoplasma capsulatum: Preparation of antisera and location of these antigens in yeast-phase cells, Infect. Immun. 14: 826–831.PubMedGoogle Scholar
  12. Green, J. H. and Pine, L., 1985, Preparation of h and m antigens of Histoplasma capsulatum free of heterologous antigens. Current Microbiol. 12: 209–216.CrossRefGoogle Scholar
  13. Greene, C. H., De Lalla, L. S., and Tompkins, V. N., 1960, Separation of specific antigens of Histoplasma capsulatum by ion-exchange chromatography, Proc. Soc. Exper. Biol. Med. 105: 140–141.Google Scholar
  14. Guerrant, G. 0. and Moss, C. W. 1984, Determination of monosaccharides as aldononitrile, O-methyloxime, alditol, and cyclitol acetate derivatives by gas chromatography, Analyt. Chem. 56: 633–638.CrossRefGoogle Scholar
  15. Heiner, D. C., 1958, Diagnosis of histoplasmosis using precipitin reactions in agar gel. Pediatrics, 22: 616–629.PubMedGoogle Scholar
  16. Kaufman, L. and Reiss, E., 1986, Serodiagnosis of fungal diseases, pp. 446–466, in: “Manual of Clinical Laboratory Immunology, 3rd ed.,” N. R. Rose, H. Friedman, and J. L. Fahey, eds. Amer. Soc. Microbiol., Washington D. C.Google Scholar
  17. Pine, L., Gross, H., Bradley-Malcolm, G., George, J. R., Gray, S. B., and Moss, C. N., 1977, Procedures for the production and separation of h and m antigens in histoplasmin: Chemical and serological properties of the isolated products, Mycopathol. 61: 131–141.CrossRefGoogle Scholar
  18. Pine, L., Smith, S. J., Gross, H., Barbaree, J. M., and Malcolm, G. B., 1981, Studies on the thermal degradation of the h and m antigens of lyophilized histoplasmin, Sabouraudia, 19: 55–70.PubMedCrossRefGoogle Scholar
  19. Prior, J. A. and Saslaw, S., 1952, Effect of repeated histoplasmin skin tests on skin reactivity and collodion agglutination, Amer. Rev. Tuberc. 66:588.Google Scholar
  20. Reiss, E., 1986, Histoplasma capsulatum, pp 77–102, in: “Molecular Immunology of Mycotic and Actinomycotic Infections”, Elsevier Science Publishing Co., Inc., New York.Google Scholar
  21. Reiss, E., Knowles, J. B., Bragg, S. L., and Kaufman, L., 1986, Monoclonal antibodies against the M-protein and carbohydrate antigens of histoplasmin characterized by the enzyme-linked immunoelectrotransfer blot method. Infect. Immun. 53: 540–546.PubMedGoogle Scholar
  22. Reiss, E., Miller, S. E.. Kaplan, W. and Kaufman, L., 1977, Antigenic, chemical, and structural properties of cell walls of Histoplasma capsulatum yeast-form chemotypes 1 and 2 after serial enzymatic hydrolysis. Infect. Immun. 16: 690–700PubMedGoogle Scholar
  23. Reiss, E., Mitchell, W. 0., Stone, S. H., and Hasenclever, H. F., 1974, Cellular immune activity of a galactomannan-protein complex from mycelia of Histoplasma capsulatum, Infect. Immun. 10: 802–809.PubMedGoogle Scholar
  24. Scalarone, G. M., Restrepo, M. A., Shacht, L. R., Rupp, G. H., and Le Roy, M. D., 1985, Lack of antibody induction by Histolyn-CYL, a new skin-testing reagent for histoplasmosis. J. Med. Vet. Mycol. 23: 335–338.CrossRefGoogle Scholar
  25. Schwarz, J., 1981, “Histoplasmosis”, Praeger Publishers, New York, pp. 151–154Google Scholar
  26. Shaw, L. W., Howell, A. Jr., and Weiss, E. S., 1950, Biological assay of lots of histoplasmin and the selection of a new working lot. Pub. Health. Rep. 65: 583–609.Google Scholar
  27. Smith, R. L. and Gilkerson, E., 1979, Quantitation of glycosaminoglycan hexosamine using 3-methyl-2-benzthiazolone hydrazone hydrochloride, Analyt. Biochem. 98: 478–480.PubMedCrossRefGoogle Scholar
  28. Smith, C. E., Whiting, E. G., Baker, E. E., Rosenberger, H. G., Beard, R. R., and Saito, M. T., 1948, The use of coccidioidin, Amer. Rev. Tuberc. 57: 330–360.Google Scholar
  29. So, L. L., and Goldstein, I. J., 1968, Protein-carbohydrate interaction XIII, The interaction of concanavalin A with α-mannans from a variety of microorganisms, J. Biol. Chem. 243: 2003–2007.PubMedGoogle Scholar
  30. Standard, P. G., and Kaufman, L., 1982, Safety considerations in handling exoantigen extracts from pathogenic fungi, J. Clin. Microbiol. 15: 663–667.PubMedGoogle Scholar
  31. Standard, P. G., Kaufman, L., and Whaley, S. P., 1984, Exoantigen test — Rapid identification of pathogenic mould isolates by immunodiffusion, in: “Immunology Series No. 11,” Procedural Guide, U. S. Centers for Disease Control, Atlanta, GA.Google Scholar
  32. Sweet, G. H., Cimprich, R. S., Cook, A. C. and Sweet, D. E., 1979, Antibodies in histoplasmosis detected by use of yeast and mycelial antigens in immunodiffusion and electroimmunodiffusion, Amer. Rev. Resp. Dis. 129: 441–449.Google Scholar
  33. Tompkins, V. N., 1965, Soluble antigenic constituents of yeast phase Histoplasma capsulatum, Amer. Rev. Resp. Dis. Suppl. 92: 126–133.Google Scholar
  34. Tsang, V. C. W., Peralta, J., and Simons, A. R., 1983, Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis, Meth. Enzymol. 92:377–391.PubMedCrossRefGoogle Scholar
  35. Utz, J. P., Becker, A., Buechner, H. A., Campbell, G. D., Einstein, H. E., and Seabury, J. H., 1976, The pulmonary mycoses: diagnostic and therapeutic guidelines, pp. 1–28, E. R. Squibb and Sons, Inc., Princeton, NJ.Google Scholar
  36. Weeke, B., 1973, Rocket Immunoelectrophoresis, Scand. J. Immunol. Suppl. 1: 37–46.PubMedCrossRefGoogle Scholar
  37. Wheat, J. French, M. L. V., Kohler, R. B., Zimmerman, S. E., Smith, W. R., Norton, J. A., Eitzen, H. E., Smith, C. D., and Slama, T. G., 1982. The diagnostic laboratory tests for histoplasmosis -analysis of experience in a large urban outbreak, Ann. Intern. Med. 97: 680–685.PubMedGoogle Scholar
  38. Wheat, L. J., Kohler, R. B., and Tewari, R. P., 1986, Diagnosis of disseminated histoplasmosis by detection of Histoplasma capsulatum antigen in serum and urine specimens, N. Engl. J. Med. 314: 83–88.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Errol Reiss
    • 1
  • Sandra L. Bragg
    • 1
  1. 1.Division of Mycotic Diseases, Center for Infectious Diseases, Centers for Disease Control, Public Health ServiceUS Department of Health and Human ServicesAtlantaUSA

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