Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1993 May;61(5):1835–1845. doi: 10.1128/iai.61.5.1835-1845.1993

The mycobacterial secreted antigen 85 complex possesses epitopes that are differentially expressed in human leprosy lesions and Mycobacterium leprae-infected armadillo tissues.

A Rambukkana 1, J D Burggraaf 1, W R Faber 1, M Harboe 1, P Teeling 1, S Krieg 1, P K Das 1
PMCID: PMC280773  PMID: 7682995

Abstract

The granulomatous skin lesions in leprosy are thought to be initiated by the immune response to certain antigens of the causative agent, Mycobacterium leprae. The antigen 85 complex is one of the major targets in the immune response to M. leprae infection. In the present study, a panel of previously characterized monoclonal antibodies (MAbs) (3A8, Rb2, A4g4, A2h11, Pe12, and A3c12) reacting with different epitopes of the 85 complex proteins of Mycobacterium tuberculosis and M. leprae was employed in a comparative immunohistological analysis to demonstrate the in situ expression of 85 complex antigenic epitopes in leprosy lesions across the clinical spectrum and in M. leprae-infected armadillo liver tissues. These MAbs showed a heterogeneous staining pattern in a given leprosy lesion. In highly bacilliferous borderline and lepromatous leprosy lesions, MAbs Rb2, A4g4, A2h11, and Pe12 stained clear rod-shaped M. leprae bacilli within macrophages, and the degree of staining correlated with the bacillary index of the lesion. On the other hand, MAbs 3A8 and A3c12 staining was mostly seen as a diffuse staining pattern within interstitial spaces and on the membranes of the infiltrated cells but not the bacilli. In paucibacillary borderline and tuberculoid leprosy lesions, only 3A8, Rb2, and A3c12 showed distinct staining in association with infiltrates in the granuloma. None of these MAbs showed any detectable reaction with control nonleprosy skin lesions, while MAb A3c12 positively stained the granulomas of both leprosy and control specimens. In situ reactivity of these MAbs with M. leprae-infected armadillo liver tissues also showed a heterogeneous staining pattern. Interestingly, a clear difference in expression of these epitopes was observed between armadillo tissues and human leprosy lesions. By immunogold ultracytochemistry, we further showed the differential localization of these MAb-reactive epitopes on the cell surface, in the cytosol, and at the vicinity of M. leprae within Kupffer cells of armadillo liver tissues. Our results indicate that these antigenic epitopes of the antigen 85 complex are differentially expressed in leprosy lesions and infected armadillo tissues and that they could be target determinants in the immunopathological responses during M. leprae infection.

Full text

PDF
1835

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abou-Zeid C., Ratliff T. L., Wiker H. G., Harboe M., Bennedsen J., Rook G. A. Characterization of fibronectin-binding antigens released by Mycobacterium tuberculosis and Mycobacterium bovis BCG. Infect Immun. 1988 Dec;56(12):3046–3051. doi: 10.1128/iai.56.12.3046-3051.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersen P., Askgaard D., Ljungqvist L., Bentzon M. W., Heron I. T-cell proliferative response to antigens secreted by Mycobacterium tuberculosis. Infect Immun. 1991 Apr;59(4):1558–1563. doi: 10.1128/iai.59.4.1558-1563.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Das P. K., Rambukkana A., Baas J. G., Groothuis D. G., Halperin M. Enzyme-linked immunosorbent assay for distinguishing serological responses of lepromatous and tuberculoid leprosies to the 29/33-kilodalton doublet and 64-kilodalton antigens of Mycobacterium tuberculosis. J Clin Microbiol. 1990 Feb;28(2):379–382. doi: 10.1128/jcm.28.2.379-382.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Espitia C., Sciutto E., Bottasso O., González-Amaro R., Hernández-Pando R., Mancilla R. High antibody levels to the mycobacterial fibronectin-binding antigen of 30-31 kD in tuberculosis and lepromatous leprosy. Clin Exp Immunol. 1992 Mar;87(3):362–367. doi: 10.1111/j.1365-2249.1992.tb03003.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Harboe M., Wiker H. G., Nagai S. Protein antigens of mycobacteria studied by quantitative immunologic techniques. Clin Infect Dis. 1992 Jan;14(1):313–319. doi: 10.1093/clinids/14.1.313. [DOI] [PubMed] [Google Scholar]
  6. Karat A. B., Job C. K., Rao P. S. Liver in leprosy: histological and biochemical findings. Br Med J. 1971 Feb 6;1(5744):307–310. doi: 10.1136/bmj.1.5744.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Khanolkar S. R., Mackenzie C. D., Lucas S. B., Hussen A., Girdhar B. K., Katoch K., McAdam K. P. Identification of Mycobacterium leprae antigens in tissues of leprosy patients using monoclonal antibodies. Int J Lepr Other Mycobact Dis. 1989 Sep;57(3):652–658. [PubMed] [Google Scholar]
  8. Modlin R. L., Melancon-Kaplan J., Young S. M., Pirmez C., Kino H., Convit J., Rea T. H., Bloom B. R. Learning from lesions: patterns of tissue inflammation in leprosy. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1213–1217. doi: 10.1073/pnas.85.4.1213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mshana R. N., Belehu A., Stoner G. L., Harboe M., Haregewoin A. Demonstration of mycobacterial antigens in leprosy tissues. Int J Lepr Other Mycobact Dis. 1982 Mar;50(1):1–10. [PubMed] [Google Scholar]
  10. Mshana R. N., Humber D. P., Harboe M., Belehu A. Demonstration of mycobacterial antigens in nerve biopsies from leprosy patients using peroxidase-antiperoxidase immunoenzyme technique. Clin Immunol Immunopathol. 1983 Dec;29(3):359–368. doi: 10.1016/0090-1229(83)90039-9. [DOI] [PubMed] [Google Scholar]
  11. Myrvang B., Godal T., Ridley D. S., Fröland S. S., Song Y. K. Immune responsiveness to Mycobacterium leprae and other mycobacterial antigens throughout the clinical and histopathological spectrum of leprosy. Clin Exp Immunol. 1973 Aug;14(4):541–553. [PMC free article] [PubMed] [Google Scholar]
  12. Naafs B., Kolk A. H., Chin A Lien R. A., Faber W. R., Van Dijk G., Kuijper S., Stolz E., Van Joost T. Anti-Mycobacterium leprae monoclonal antibodies cross-react with human skin: an alternative explanation for the immune responses in leprosy. J Invest Dermatol. 1990 May;94(5):685–688. doi: 10.1111/1523-1747.ep12876264. [DOI] [PubMed] [Google Scholar]
  13. Nagai S., Wiker H. G., Harboe M., Kinomoto M. Isolation and partial characterization of major protein antigens in the culture fluid of Mycobacterium tuberculosis. Infect Immun. 1991 Jan;59(1):372–382. doi: 10.1128/iai.59.1.372-382.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Narayanan R. B. Immunopathology of leprosy granulomas--current status: a review. Lepr Rev. 1988 Mar;59(1):75–82. [PubMed] [Google Scholar]
  15. Narayanan R. B., Ramu G., Sinha S., Sengupta U., Malaviya G. N., Desikan K. V. Demonstration of Mycobacterium leprae specific antigens in leprosy lesions using monoclonal antibodies. Indian J Lepr. 1985 Apr-Jun;57(2):258–264. [PubMed] [Google Scholar]
  16. Pessolani M. C., Rumjanek F. D., Marques M. A., de Melo F. S., Sarno E. N. Serological response of patients with leprosy to a 28- to 30-kilodalton protein doublet from early cultures of Mycobacterium bovis BCG. J Clin Microbiol. 1989 Oct;27(10):2184–2189. doi: 10.1128/jcm.27.10.2184-2189.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rambukkana A., Das P. K., Burggraaf J. D., Faber W. R., Teeling P., Krieg S., Thole J. E., Harboe M. Identification and characterization of epitopes shared between the mycobacterial 65-kilodalton heat shock protein and the actively secreted antigen 85 complex: their in situ expression on the cell wall surface of Mycobacterium leprae. Infect Immun. 1992 Nov;60(11):4517–4527. doi: 10.1128/iai.60.11.4517-4527.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rambukkana A., Das P. K., Burggraaf J. D., Yong S., Faber W. R., Thole J. E., Harboe M. Heterogeneity of monoclonal antibody-reactive epitopes on mycobacterial 30-kilodalton-region proteins and the secreted antigen 85 complex and demonstration of antigen 85B on the Mycobacterium leprae cell wall surface. Infect Immun. 1992 Dec;60(12):5172–5181. doi: 10.1128/iai.60.12.5172-5181.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rambukkana A., Das P. K., Chand A., Baas J. G., Groothuis D. G., Kolk A. H. Subcellular distribution of monoclonal antibody defined epitopes on immunodominant Mycobacterium tuberculosis proteins in the 30-kDa region: identification and localization of 29/33-kDa doublet proteins on mycobacterial cell wall. Scand J Immunol. 1991 Jun;33(6):763–775. doi: 10.1111/j.1365-3083.1991.tb02551.x. [DOI] [PubMed] [Google Scholar]
  20. Rambukkana A., Das P. K., Krieg S., Faber W. R. Association of the mycobacterial 30-kDa region proteins with the cutaneous infiltrates of leprosy lesions. Evidence for the involvement of the major mycobacterial secreted proteins in the local immune response of leprosy. Scand J Immunol. 1992 Jul;36(1):35–48. doi: 10.1111/j.1365-3083.1992.tb02938.x. [DOI] [PubMed] [Google Scholar]
  21. Ridley D. S., Jopling W. H. Classification of leprosy according to immunity. A five-group system. Int J Lepr Other Mycobact Dis. 1966 Jul-Sep;34(3):255–273. [PubMed] [Google Scholar]
  22. Ridley D. S. The pathogenesis of the early skin lesion in leprosy. J Pathol. 1973 Nov;111(3):191–206. doi: 10.1002/path.1711110307. [DOI] [PubMed] [Google Scholar]
  23. Rumschlag H. S., Shinnick T. M., Cohen M. L. Serological responses of patients with lepromatous and tuberculoid leprosy to 30-, 31-, and 32-kilodalton antigens of Mycobacterium tuberculosis. J Clin Microbiol. 1988 Oct;26(10):2200–2202. doi: 10.1128/jcm.26.10.2200-2202.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Thole J. E., Schöningh R., Janson A. A., Garbe T., Cornelisse Y. E., Clark-Curtiss J. E., Kolk A. H., Ottenhoff T. H., De Vries R. R., Abou-Zeid C. Molecular and immunological analysis of a fibronectin-binding protein antigen secreted by Mycobacterium leprae. Mol Microbiol. 1992 Jan;6(2):153–163. doi: 10.1111/j.1365-2958.1992.tb01996.x. [DOI] [PubMed] [Google Scholar]
  25. Van Vooren J. P., Drowart A., De Bruyn J., Launois P., Millan J., Delaporte E., Develoux M., Yernault J. C., Huygen K. Humoral responses against the 85A and 85B antigens of Mycobacterium bovis BCG in patients with leprosy and tuberculosis. J Clin Microbiol. 1992 Jun;30(6):1608–1610. doi: 10.1128/jcm.30.6.1608-1610.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Van Voorhis W. C., Kaplan G., Sarno E. N., Horwitz M. A., Steinman R. M., Levis W. R., Nogueira N., Hair L. S., Gattass C. R., Arrick B. A. The cutaneous infiltrates of leprosy: cellular characteristics and the predominant T-cell phenotypes. N Engl J Med. 1982 Dec 23;307(26):1593–1597. doi: 10.1056/NEJM198212233072601. [DOI] [PubMed] [Google Scholar]
  27. Van der Loos C. M., Das P. K., Van den Oord J. J., Houthoff H. J. Multiple immunoenzyme staining techniques. Use of fluoresceinated, biotinylated and unlabelled monoclonal antibodies. J Immunol Methods. 1989 Feb 8;117(1):45–52. doi: 10.1016/0022-1759(89)90117-8. [DOI] [PubMed] [Google Scholar]
  28. Watson J. D. Leprosy: understanding protective immunity. Immunol Today. 1989 Jul;10(7):218–221. doi: 10.1016/0167-5699(89)90253-3. [DOI] [PubMed] [Google Scholar]
  29. Wiker H. G., Harboe M., Nagai S., Bennedsen J. Quantitative and qualitative studies on the major extracellular antigen of Mycobacterium tuberculosis H37Rv and Mycobacterium bovis BCG. Am Rev Respir Dis. 1990 Apr;141(4 Pt 1):830–838. doi: 10.1164/ajrccm/141.4_Pt_1.830. [DOI] [PubMed] [Google Scholar]
  30. Wiker H. G., Harboe M., Nagai S., Patarroyo M. E., Ramirez C., Cruz N. MPB59, a widely cross-reacting protein of Mycobacterium bovis BCG. Int Arch Allergy Appl Immunol. 1986;81(4):307–314. doi: 10.1159/000234154. [DOI] [PubMed] [Google Scholar]
  31. Wiker H. G., Nagai S., Harboe M., Ljungqvist L. A family of cross-reacting proteins secreted by Mycobacterium tuberculosis. Scand J Immunol. 1992 Aug;36(2):307–319. doi: 10.1111/j.1365-3083.1992.tb03104.x. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES