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. 1997 Jan;4(1):49–56. doi: 10.1128/cdli.4.1.49-56.1997

Human humoral responses to antigens of Mycobacterium tuberculosis: immunodominance of high-molecular-mass antigens.

S Laal 1, K M Samanich 1, M G Sonnenberg 1, S Zolla-Pazner 1, J M Phadtare 1, J T Belisle 1
PMCID: PMC170474  PMID: 9008280

Abstract

The selection of antigens of Mycobacterium tuberculosis for most studies of humoral responses in tuberculosis patients has been restricted to molecules that were either immunodominant in immunized animals or amenable to biochemical purification rather than those that were reactive with the human immune system. Delineation of antigens that elicit humoral responses during the natural course of disease progression in humans has been hindered by the presence of cross-reactive antibodies to conserved regions on ubiquitous prokaryotic antigens in sera from healthy individuals and tuberculosis patients. The levels of cross-reactive antibodies in the sera were reduced by preadsorption with Escherichia coli lysates, prior to studying their reactivity against a large panel of M. tuberculosis antigens to which the human immune system may be exposed during natural infection and disease. Thus, reactivity against pools of secreted, cellular, and cell wall-associated antigens of M. tuberculosis was assessed by an enzyme-linked immunosorbent assay (ELISA). Initial results suggested that the secreted protein preparation contained antigens most frequently recognized by the humoral responses of pulmonary tuberculosis patients. The culture filtrate proteins were subsequently size fractionated by preparative polyacrylamide gel electrophoresis, characterized by reaction with murine monoclonal antibodies to known antigens of M. tuberculosis by an ELISA, and assessed for reactivity with tuberculous and nontuberculous sera. Results show that a secreted antigen of 88 kDa elicits a strong antibody response in a high percentage of patients with pulmonary tuberculosis. This and other antigens identified on the basis of their reactivity with patient sera may prove useful for developing serodiagnosis for tuberculosis.

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Selected References

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  1. Andersen A. B., Andersen P., Ljungqvist L. Structure and function of a 40,000-molecular-weight protein antigen of Mycobacterium tuberculosis. Infect Immun. 1992 Jun;60(6):2317–2323. doi: 10.1128/iai.60.6.2317-2323.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersen A. B., Hansen E. B. Structure and mapping of antigenic domains of protein antigen b, a 38,000-molecular-weight protein of Mycobacterium tuberculosis. Infect Immun. 1989 Aug;57(8):2481–2488. doi: 10.1128/iai.57.8.2481-2488.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Andersen P., Askgaard D., Gottschau A., Bennedsen J., Nagai S., Heron I. Identification of immunodominant antigens during infection with Mycobacterium tuberculosis. Scand J Immunol. 1992 Dec;36(6):823–831. doi: 10.1111/j.1365-3083.1992.tb03144.x. [DOI] [PubMed] [Google Scholar]
  4. Bardana E. J., Jr, McClatchy J. K., Farr R. S., Minden P. Universal occurrence of antibodies to tubercle bacilli in sera from non-tuberculous and tuberculous individuals. Clin Exp Immunol. 1973 Jan;13(1):65–77. [PMC free article] [PubMed] [Google Scholar]
  5. Bothamley G. H., Rudd R., Festenstein F., Ivanyi J. Clinical value of the measurement of Mycobacterium tuberculosis specific antibody in pulmonary tuberculosis. Thorax. 1992 Apr;47(4):270–275. doi: 10.1136/thx.47.4.270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bothamley G., Udani P., Rudd R., Festenstein F., Ivanyi J. Humoral response to defined epitopes of tubercle bacilli in adult pulmonary and child tuberculosis. Eur J Clin Microbiol Infect Dis. 1988 Oct;7(5):639–645. doi: 10.1007/BF01964242. [DOI] [PubMed] [Google Scholar]
  7. Braibant M., De Wit L., Peirs P., Kalai M., Ooms J., Drowart A., Huygen K., Content J. Structure of the Mycobacterium tuberculosis antigen 88, a protein related to the Escherichia coli PstA periplasmic phosphate permease subunit. Infect Immun. 1994 Mar;62(3):849–854. doi: 10.1128/iai.62.3.849-854.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Calle J. M., Nardin E. H., Clavijo P., Boudin C., Stüber D., Takacs B., Nussenzweig R. S., Cochrane A. H. Recognition of different domains of the Plasmodium falciparum CS protein by the sera of naturally infected individuals compared with those of sporozoite-immunized volunteers. J Immunol. 1992 Oct 15;149(8):2695–2701. [PubMed] [Google Scholar]
  9. Carlin N. I., Löfdahl S., Magnusson M. Monoclonal antibodies specific for elongation factor Tu and complete nucleotide sequence of the tuf gene in Mycobacterium tuberculosis. Infect Immun. 1992 Aug;60(8):3136–3142. doi: 10.1128/iai.60.8.3136-3142.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chan S. L., Reggiardo Z., Daniel T. M., Girling D. J., Mitchison D. A. Serodiagnosis of tuberculosis using an ELISA with antigen 5 and a hemagglutination assay with glycolipid antigens. Results in patients with newly diagnosed pulmonary tuberculosis ranging in extent of disease from minimal to extensive. Am Rev Respir Dis. 1990 Aug;142(2):385–389. doi: 10.1164/ajrccm/142.2.385. [DOI] [PubMed] [Google Scholar]
  11. Daffe M., Brennan P. J., McNeil M. Predominant structural features of the cell wall arabinogalactan of Mycobacterium tuberculosis as revealed through characterization of oligoglycosyl alditol fragments by gas chromatography/mass spectrometry and by 1H and 13C NMR analyses. J Biol Chem. 1990 Apr 25;265(12):6734–6743. [PubMed] [Google Scholar]
  12. Daniel T. M., De Murillo G. L., Sawyer J. A., Griffin A. M., Pinto E., Debanne S. M., Espinosa P., Cespedes E. Field evaluation of enzyme-linked immunosorbent assay for the serodiagnosis of tuberculosis. Am Rev Respir Dis. 1986 Oct;134(4):662–665. doi: 10.1164/arrd.1986.134.4.662. [DOI] [PubMed] [Google Scholar]
  13. Daniel T. M., Debanne S. M. The serodiagnosis of tuberculosis and other mycobacterial diseases by enzyme-linked immunosorbent assay. Am Rev Respir Dis. 1987 May;135(5):1137–1151. doi: 10.1164/arrd.1987.135.5.1137. [DOI] [PubMed] [Google Scholar]
  14. Daniel T. M., Debanne S. M., van der Kuyp F. Enzyme-linked immunosorbent assay using Mycobacterium tuberculosis antigen 5 and PPD for the serodiagnosis of tuberculosis. Chest. 1985 Sep;88(3):388–392. doi: 10.1378/chest.88.3.388. [DOI] [PubMed] [Google Scholar]
  15. Das S., Cheng S. H., Lowrie D. B., Walker K. B., Mitchison D. A., Vallishayee R. S., Narayanan P. R. The pattern of mycobacterial antigen recognition in sera from Mantoux-negative individuals is essentially unaffected by bacille Calmette-Guérin (BCG) vaccination in either south India or London. Clin Exp Immunol. 1992 Sep;89(3):402–406. doi: 10.1111/j.1365-2249.1992.tb06970.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Davenport M. P., McKenzie K. R., Basten A., Britton W. J. The variable C-terminal region of the Mycobacterium leprae 70-kilodalton heat shock protein is the target for humoral immune responses. Infect Immun. 1992 Mar;60(3):1170–1177. doi: 10.1128/iai.60.3.1170-1177.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Del Giudice G., Gervaix A., Costantino P., Wyler C. A., Tougne C., de Graeff-Meeder E. R., van Embden J., van der Zee R., Nencioni L., Rappuoli R. Priming to heat shock proteins in infants vaccinated against pertussis. J Immunol. 1993 Mar 1;150(5):2025–2032. [PubMed] [Google Scholar]
  18. Drowart A., Huygen K., De Bruyn J., Yernault J. C., Farber C. M., Van Vooren J. P. Antibody levels to whole culture filtrate antigens and to purified P32 during treatment of smear-positive tuberculosis. Chest. 1991 Sep;100(3):685–687. doi: 10.1378/chest.100.3.685. [DOI] [PubMed] [Google Scholar]
  19. Espitia C., Cervera I., González R., Mancilla R. A 38-kD Mycobacterium tuberculosis antigen associated with infection. Its isolation and serologic evaluation. Clin Exp Immunol. 1989 Sep;77(3):373–377. [PMC free article] [PubMed] [Google Scholar]
  20. Garsia R. J., Hellqvist L., Booth R. J., Radford A. J., Britton W. J., Astbury L., Trent R. J., Basten A. Homology of the 70-kilodalton antigens from Mycobacterium leprae and Mycobacterium bovis with the Mycobacterium tuberculosis 71-kilodalton antigen and with the conserved heat shock protein 70 of eucaryotes. Infect Immun. 1989 Jan;57(1):204–212. doi: 10.1128/iai.57.1.204-212.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Grandia A. A., de Visser H., van Embden J. D., van der Zee R., van den Berg W. B., Hazenberg M. P. Natural antibodies to 65 kD mycobacterial heat shock protein in rats do not correlate with susceptibility for Mycobacterium tuberculosis induced adjuvant arthritis. Immunobiology. 1991 Mar;182(2):127–134. doi: 10.1016/S0171-2985(11)80196-8. [DOI] [PubMed] [Google Scholar]
  22. Grange J. M. The humoral immune response in tuberculosis: its nature, biological role and diagnostic usefulness. Adv Tuberc Res. 1984;21:1–78. [PubMed] [Google Scholar]
  23. Harboe M., Wiker H. G. The 38-kDa protein of Mycobacterium tuberculosis: a review. J Infect Dis. 1992 Oct;166(4):874–884. doi: 10.1093/infdis/166.4.874. [DOI] [PubMed] [Google Scholar]
  24. Hartskeerl R. A., van Rens R. M., Stabel L. F., de Wit M. Y., Klatser P. R. Selection and characterization of recombinant clones that produce Mycobacterium leprae antigens recognized by antibodies in sera from household contacts of leprosy patients. Infect Immun. 1990 Sep;58(9):2821–2827. doi: 10.1128/iai.58.9.2821-2827.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Havlir D. V., Wallis R. S., Boom W. H., Daniel T. M., Chervenak K., Ellner J. J. Human immune response to Mycobacterium tuberculosis antigens. Infect Immun. 1991 Feb;59(2):665–670. doi: 10.1128/iai.59.2.665-670.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Hirschfield G. R., McNeil M., Brennan P. J. Peptidoglycan-associated polypeptides of Mycobacterium tuberculosis. J Bacteriol. 1990 Feb;172(2):1005–1013. doi: 10.1128/jb.172.2.1005-1013.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ivanyi J., Bothamley G. H., Jackett P. S. Immunodiagnostic assays for tuberculosis and leprosy. Br Med Bull. 1988 Jul;44(3):635–649. doi: 10.1093/oxfordjournals.bmb.a072273. [DOI] [PubMed] [Google Scholar]
  28. Ivanyi J., Krambovitis E., Keen M. Evaluation of a monoclonal antibody (TB72) based serological test for tuberculosis. Clin Exp Immunol. 1983 Nov;54(2):337–345. [PMC free article] [PubMed] [Google Scholar]
  29. Jackett P. S., Bothamley G. H., Batra H. V., Mistry A., Young D. B., Ivanyi J. Specificity of antibodies to immunodominant mycobacterial antigens in pulmonary tuberculosis. J Clin Microbiol. 1988 Nov;26(11):2313–2318. doi: 10.1128/jcm.26.11.2313-2318.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Khanolkar-Young S., Kolk A. H., Andersen A. B., Bennedsen J., Brennan P. J., Rivoire B., Kuijper S., McAdam K. P., Abe C., Batra H. V. Results of the third immunology of leprosy/immunology of tuberculosis antimycobacterial monoclonal antibody workshop. Infect Immun. 1992 Sep;60(9):3925–3927. doi: 10.1128/iai.60.9.3925-3927.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Laal S., Sharma Y. D., Prasad H. K., Murtaza A., Singh S., Tangri S., Misra R. S., Nath I. Recombinant fusion protein identified by lepromatous sera mimics native Mycobacterium leprae in T-cell responses across the leprosy spectrum. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):1054–1058. doi: 10.1073/pnas.88.3.1054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ma Y., Wang Y. M., Daniel T. M. Enzyme-linked immunosorbent assay using Mycobacterium tuberculosis antigen 5 for the diagnosis of pulmonary tuberculosis in China. Am Rev Respir Dis. 1986 Dec;134(6):1273–1275. doi: 10.1164/arrd.1986.134.5.1273. [DOI] [PubMed] [Google Scholar]
  33. Meeker H. C., Williams D. L., Anderson D. C., Gillis T. P., Schuller-Levis G., Levis W. R. Analysis of human antibody epitopes on the 65-kilodalton protein of Mycobacterium leprae by using synthetic peptides. Infect Immun. 1989 Dec;57(12):3689–3694. doi: 10.1128/iai.57.12.3689-3694.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Orme I. M., Andersen P., Boom W. H. T cell response to Mycobacterium tuberculosis. J Infect Dis. 1993 Jun;167(6):1481–1497. doi: 10.1093/infdis/167.6.1481. [DOI] [PubMed] [Google Scholar]
  35. Raja A., Narayanan P. R., Mathew R., Prabhakar R. Characterization of mycobacterial antigens and antibodies in circulating immune complexes from pulmonary tuberculosis. J Lab Clin Med. 1995 May;125(5):581–587. [PubMed] [Google Scholar]
  36. Raviglione M. C., Snider D. E., Jr, Kochi A. Global epidemiology of tuberculosis. Morbidity and mortality of a worldwide epidemic. JAMA. 1995 Jan 18;273(3):220–226. [PubMed] [Google Scholar]
  37. Results of a World Health Organization-sponsored workshop to characterize antigens recognized by mycobacterium-specific monoclonal antibodies. Infect Immun. 1986 Feb;51(2):718–720. doi: 10.1128/iai.51.2.718-720.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sada E., Brennan P. J., Herrera T., Torres M. Evaluation of lipoarabinomannan for the serological diagnosis of tuberculosis. J Clin Microbiol. 1990 Dec;28(12):2587–2590. doi: 10.1128/jcm.28.12.2587-2590.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sada E., Ferguson L. E., Daniel T. M. An ELISA for the serodiagnosis of tuberculosis using a 30,000-Da native antigen of Mycobacterium tuberculosis. J Infect Dis. 1990 Oct;162(4):928–931. doi: 10.1093/infdis/162.4.928. [DOI] [PubMed] [Google Scholar]
  40. Shinnick T. M., Plikaytis B. B., Hyche A. D., Van Landingham R. M., Walker L. L. The Mycobacterium tuberculosis BCG-a protein has homology with the Escherichia coli GroES protein. Nucleic Acids Res. 1989 Feb 11;17(3):1254–1254. doi: 10.1093/nar/17.3.1254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Shinnick T. M., Vodkin M. H., Williams J. C. The Mycobacterium tuberculosis 65-kilodalton antigen is a heat shock protein which corresponds to common antigen and to the Escherichia coli GroEL protein. Infect Immun. 1988 Feb;56(2):446–451. doi: 10.1128/iai.56.2.446-451.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Thole J. E., Keulen W. J., De Bruyn J., Kolk A. H., Groothuis D. G., Berwald L. G., Tiesjema R. H., van Embden J. D. Characterization, sequence determination, and immunogenicity of a 64-kilodalton protein of Mycobacterium bovis BCG expressed in escherichia coli K-12. Infect Immun. 1987 Jun;55(6):1466–1475. doi: 10.1128/iai.55.6.1466-1475.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Udaykumar, Saxena R. K. Antigenic epitopes on Mycobacterium tuberculosis recognized by antibodies in tuberculosis and mouse antisera. FEMS Microbiol Immunol. 1991 Feb;3(1):7–12. doi: 10.1111/j.1574-6968.1991.tb04156.x. [DOI] [PubMed] [Google Scholar]
  44. Van Vooren J. P., Drowart A., De Cock M., Van Onckelen A., D'Hoop M. H., Yernault J. C., Valcke C., Huygen K. Humoral immune response of tuberculous patients against the three components of the Mycobacterium bovis BCG 85 complex separated by isoelectric focusing. J Clin Microbiol. 1991 Oct;29(10):2348–2350. doi: 10.1128/jcm.29.10.2348-2350.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Verbon A. Development of a serological test for tuberculosis. Problems and potential. Trop Geogr Med. 1994;46(5):275–279. [PubMed] [Google Scholar]
  46. Verbon A., Kuijper S., Jansen H. M., Speelman P., Kolk A. H. Antigens in culture supernatant of Mycobacterium tuberculosis: epitopes defined by monoclonal and human antibodies. J Gen Microbiol. 1990 May;136(5):955–964. doi: 10.1099/00221287-136-5-955. [DOI] [PubMed] [Google Scholar]
  47. Wallis R. S., Paranjape R., Phillips M. Identification by two-dimensional gel electrophoresis of a 58-kilodalton tumor necrosis factor-inducing protein of Mycobacterium tuberculosis. Infect Immun. 1993 Feb;61(2):627–632. doi: 10.1128/iai.61.2.627-632.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wieles B., Nagai S., Wiker H. G., Harboe M., Ottenhoff T. H. Identification and functional characterization of thioredoxin of Mycobacterium tuberculosis. Infect Immun. 1995 Dec;63(12):4946–4948. doi: 10.1128/iai.63.12.4946-4948.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Young D. B., Kaufmann S. H., Hermans P. W., Thole J. E. Mycobacterial protein antigens: a compilation. Mol Microbiol. 1992 Jan;6(2):133–145. doi: 10.1111/j.1365-2958.1992.tb01994.x. [DOI] [PubMed] [Google Scholar]
  50. Zhang Y., Lathigra R., Garbe T., Catty D., Young D. Genetic analysis of superoxide dismutase, the 23 kilodalton antigen of Mycobacterium tuberculosis. Mol Microbiol. 1991 Feb;5(2):381–391. doi: 10.1111/j.1365-2958.1991.tb02120.x. [DOI] [PubMed] [Google Scholar]

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