Skip to main content
Log in

In situ localization of antigens of Histoplasma capsulatum using colloidal gold immune electron microscopy

  • Published:
Mycopathologia Aims and scope Submit manuscript

Abstract

Histoplasma capsulatum contains multiple antigens, among them the H antigen and M antigen, which are useful in serologic testing for histoplasmosis. We prepared 7 mouse monoclonal antibodies (5 IgG, 2 IgM) to histoplasmin, and compared these with polyclonal histoplasmin antibodies raised in rabbits and mice. Both monoclonal and polyclonal antibodies were high titered by ELISA. Colloidal gold immune electron microscopy (CGIEM) showed that polyclonal antibodies to histoplasmin or H antigen bound at multiple sites in the cell wall, cytoplasm, and nucleus of Histoplasma yeast cells. In contrast, antibodies to M antigen selectively label the cell membrane and antibodies to alkali soluble cell wall antigen label only the cell wall. Polyclonal antibodies cross reacted extensively with other fungi, both by ELISA and CGIEM. Monoclonal antibodies stained only cytoplasmic epitopes, but also cross reacted with other fungi by electron microscopy. Only periodate treated H antigen elicited polyclonal antibodies which were more specific than those of untreated H antigen or histoplasmin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Cox RA, Larsh HW. Isolation of skin test-active preparations from yeast-phase cells of Bastomyces dermatitidis. Inf and Imm 1974; 10:42–7.

    Google Scholar 

  2. Cox RA. Cross-reactivity between antigens of Coccidioides immitis, Histoplasma capsulatum, and Blastomyces dermatitidis in lymphocyte transformation assays. Inf and Imm 1979; 25:932–8.

    Google Scholar 

  3. Graybill JR, Patino MM, Gomez AM, Ahrens J. Detection of histoplasmal antigens in mice undergoing experimental pulmonary histoplasmosis. Am Rev Resp Dis 1985; 132:752–6.

    Google Scholar 

  4. Green JH, Harrell WK, Gray SB, Johnson JE, Bolin RC, Gross HN, Malcolm GB. H and M antigens of Histoplasma capsulatum: preparaton of antisera and localization of these antigens in yeast-phase cells. Inf and Imm 1976; 14:826–31.

    Google Scholar 

  5. Heiner DG. Diagnosis of histoplasmosis using precipitin reactions in agar gel. Pediatrics 1958; 22:616–27.

    Google Scholar 

  6. Kumar BV, Medoff G, Kobayashi GS, Sieling WL. Crossreacting human and rabbit antibodies to antigens of Histoplasma capsulatum, Candida albicans, and Saccharmoyces cerevisiae. Inf and Imm 1985; 48:806–12.

    Google Scholar 

  7. Lefkovits I, Waldmann H. Limiting dilution analysis of the cells of the immune system. I. The clonal basis of the immune response. Immunology Today 1984; 5:265–8.

    Google Scholar 

  8. Pine L, Gross H, Lalcolm GB, George JR, Gray SB, Moss CW. Procedures for the production and separation of H and M antigens in histoplasmin: chemical and serological properties of the isolated products. Mycopathologia 1977; 61:131–41.

    Google Scholar 

  9. Pine L, Malcolm GB, Gross H, Gray SB. Evaluation of purified H and M antigens of histoplasmin as reagents in the complement fixation test. Sabouraudia 1978; 16:257–69.

    Google Scholar 

  10. Pine L, Fears MB, Malcolm GB. Heated histoplasmin as a control for non-specificity in the complement fixation test for histoplasmosis. Sabouraudia 1983; 21:233–7.

    Google Scholar 

  11. Pine L, Green JH, Gross H, Malcolm GB, Harrel WK. Purified histoplasmin reagents for the identification of h and m antigens and homologous antibodies by immunodiffusion or complement fixation tests. Mycopathologia 1985; 91:39–40.

    Google Scholar 

  12. Reiss E, Knowles JB, Bragg SL, Kaufman L. Monoclonal antibodies against the M-protein and carbohydrate antigens of histoplasmin characterized by the enzyme-linked immunoelectrotransfer method. Inf and Imm 1986; 53:540–6.

    Google Scholar 

  13. Wheat J, French MLV, Kohler RB, Zimmerman SE, Smith WR, Norton JA, Eitzen HE, Smith CD, Slama TG. The diagnostic tests for histoplasmosis: analysis of experience in a large urban outbreak. Ann Int Med 1982; 97:680–5.

    Google Scholar 

  14. Wiggins GL, Schubert JH. Relationship of histoplasma agargel bands and complement fixation titers in histoplasmosis. J Bacteriol 1965; 89:589–96.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Graybill, J.R., Patiño, M. & Ahrens, J. In situ localization of antigens of Histoplasma capsulatum using colloidal gold immune electron microscopy. Mycopathologia 104, 181–188 (1988). https://doi.org/10.1007/BF00437434

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00437434

Key words

Navigation