, Volume 61, Issue 3, pp 131–141 | Cite as

Procedures for the production and separation of H and M antigens in histoplasmin: Chemical and serological properties of the isolated products

  • Leo Pine
  • Herman Gross
  • Georgia Bradley Malcolm
  • James R. George
  • S. B. Gray
  • C. Wayne Moss


Two strains of Histoplasma capsulatum were required to prepare maximum yields of H and of M antigen from histoplasmin. The antigens were separated and partially purified by a series of procedures yielding an overall recovery of 70 to 90% of the individual antigens. Stable products suitable for use as reference products were obtained when the final purification step employed DEAE-cellulose with phosphate buffer elution at increasing molarity and decreasing pH. A final step of purification of each antigen with slab acrylamide gel electrophoresis gave products which were highly reactive and specific in a variety of serological tests with sera from persons with proven cases of histoplasmosis and with natural infections of heterologous deep mycoses. These antigens were maximally active at concentrations of 2 to 16 μg protein in the complement fixation, capillary precipitin, microimmunodiffusion, or immunoelectrophoresis tests; 0.5 μg gave a maximum delayed cutaneous hypersensitivity reaction in homologously infected animals and caused no appreciable reaction in control animals. Although these antigens appeared to be specific when tested with sera from persons with natural infections, the M and H antigens demonstrated the presence of an additional antigen reacting with sera of rabbits immunized with cell membrane and cell particulate fractions of Blastomyces dermatitidis. After purification by electrophoresis, both the H and M antigens of some preparations showed some decomposition and loss of reactivity after storage at 5 C for more than six months. The overall results suggest that the purified H and M antigens of Heiner (12) have multiple serological reactivity and may function in precipitin reactions, complementfixing reactions, hemagglutination of formalin-fixed goose red blood cells, and as antigens for delayed cutaneous tests.


Natural Infection Histoplasmosis Reference Product Cutaneous Test Serological Reactivity 


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Copyright information

© Dr. W. Junk b.v. 1977

Authors and Affiliations

  • Leo Pine
    • 1
  • Herman Gross
    • 1
  • Georgia Bradley Malcolm
    • 1
  • James R. George
    • 1
  • S. B. Gray
    • 1
  • C. Wayne Moss
    • 1
  1. 1.U.S. Department of Health, Education, and WelfareCenter for Disease Control, Bureau of Laboratories, Public Health ServiceAtlantaUSA

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