Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Mar;35(3):553–557. doi: 10.1128/jcm.35.3.553-557.1997

Evaluation of the recombinant 38-kilodalton antigen of Mycobacterium tuberculosis as a potential immunodiagnostic reagent.

R J Wilkinson 1, K Hasløv 1, R Rappuoli 1, F Giovannoni 1, P R Narayanan 1, C R Desai 1, H M Vordermeier 1, J Paulsen 1, G Pasvol 1, J Ivanyi 1, M Singh 1
PMCID: PMC229625  PMID: 9041387

Abstract

The diagnosis of infection caused by Mycobacterium tuberculosis is of increased public health concern following increases in the number of cases in developed countries and major increases in developing countries associated with the spread of human immunodeficiency virus (HIV) infection. The specificity of purified protein derivative skin testing for the detection of infection is compromised by exposure to environmental mycobacteria. Examination of sputum detects the most infectious patients, but not those with extrapulmonary disease. The 38-kDa antigen of M. tuberculosis contains two M. tuberculosis-specific B-cell epitopes. We overexpressed the gene for this antigen in Escherichia coli and evaluated the recombinant product in in vitro assays of T-cell function and as a target for the antibody response in humans. The sensitivity and specificity of the antigen as a skin test reagent were also assessed in outbred guinea pigs. We found that 69% of healthy sensitized humans recognize the antigen in vitro, as manifested by both cell proliferation and the production of gamma interferon. Untreated patients initially have a lower frequency of response (38%); this recovers to 72% during therapy. A total of 292 patients (20 with HIV coinfection) and 58 controls were examined for production of antibody to the 38-kDa antigen by using a commercially available kit. The sensitivity of the test in comparison with that of culture was 72.6%, and the specificity was 94.9%. The antigen was also tested for its ability to induce skin reactions in outbred guinea pigs sensitized by various mycobacterial species. The antigen provoked significant skin reactions in M. tuberculosis-, M. bovis BCG-, and M. intracellulare-sensitized animals. The significance of these findings and the usefulness of this antigen in immunodiagnosis are discussed.

Full Text

The Full Text of this article is available as a PDF (117.6 KB).

Selected References

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

  1. Asherson G. L., Dieli F., Sireci G., Salerno A. Role of IL-4 in delayed type hypersensitivity. Clin Exp Immunol. 1996 Jan;103(1):1–4. doi: 10.1046/j.1365-2249.1996.845537.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barnes P. F., Mistry S. D., Cooper C. L., Pirmez C., Rea T. H., Modlin R. L. Compartmentalization of a CD4+ T lymphocyte subpopulation in tuberculous pleuritis. J Immunol. 1989 Feb 15;142(4):1114–1119. [PubMed] [Google Scholar]
  3. 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]
  4. Chang Z., Choudhary A., Lathigra R., Quiocho F. A. The immunodominant 38-kDa lipoprotein antigen of Mycobacterium tuberculosis is a phosphate-binding protein. J Biol Chem. 1994 Jan 21;269(3):1956–1958. [PubMed] [Google Scholar]
  5. Coates A. R., Hewitt J., Allen B. W., Ivanyi J., Mitchison D. A. Antigenic diversity of Mycobacterium tuberculosis and Mycobacterium bovis detected by means of monoclonal antibodies. Lancet. 1981 Jul 25;2(8239):167–169. doi: 10.1016/s0140-6736(81)90355-x. [DOI] [PubMed] [Google Scholar]
  6. Daniel T. M., Balestrino E. A., Balestrino O. C., Davidson P. T., Debanne S. M., Kataria S., Kataria Y. P., Scocozza J. B. The tuberculin specificity in humans of Mycobacterium tuberculosis antigen 5. Am Rev Respir Dis. 1982 Oct;126(4):600–606. doi: 10.1164/arrd.1982.126.4.600. [DOI] [PubMed] [Google Scholar]
  7. Daniel T. M., Ellner J. J., Todd L. S., McCoy D. W., Payne V. D., Anderson P. A., Bhe F. T. Immunobiology and species distribution of Mycobacterium tuberculosis antigen 5. Infect Immun. 1979 Apr;24(1):77–82. doi: 10.1128/iai.24.1.77-82.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. De Smet K. A., Hellyer T. J., Khan A. W., Brown I. N., Ivanyi J. Genetic and serovar typing of clinical isolates of the Mycobacterium avium-intracellulare complex. Tuber Lung Dis. 1996 Feb;77(1):71–76. doi: 10.1016/s0962-8479(96)90079-9. [DOI] [PubMed] [Google Scholar]
  9. Dolin P. J., Raviglione M. C., Kochi A. Global tuberculosis incidence and mortality during 1990-2000. Bull World Health Organ. 1994;72(2):213–220. [PMC free article] [PubMed] [Google Scholar]
  10. Grange J. M. Infection and disease due to the environmental mycobacteria. Trans R Soc Trop Med Hyg. 1987;81(2):179–182. doi: 10.1016/0035-9203(87)90206-9. [DOI] [PubMed] [Google Scholar]
  11. Hasløv K., Andersen A. B., Ljungqvist L., Weis Bentzon M. Comparison of the immunological activity of five defined antigens from Mycobacterium tuberculosis in seven inbred guinea pig strains. The 38-kDa antigen is immunodominant. Scand J Immunol. 1990 Apr;31(4):503–514. doi: 10.1111/j.1365-3083.1990.tb02798.x. [DOI] [PubMed] [Google Scholar]
  12. Hirsch C. S., Hussain R., Toossi Z., Dawood G., Shahid F., Ellner J. J. Cross-modulation by transforming growth factor beta in human tuberculosis: suppression of antigen-driven blastogenesis and interferon gamma production. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3193–3198. doi: 10.1073/pnas.93.8.3193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Martin-Casabona N., Gonzalez Fuente T., Papa F., Rosselló Urgell J., Vidal Plá R., Codina Grau G., Ruiz Camps I. Time course of anti-SL-IV immunoglobulin G antibodies in patients with tuberculosis and tuberculosis-associated AIDS. J Clin Microbiol. 1992 May;30(5):1089–1093. doi: 10.1128/jcm.30.5.1089-1093.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McMurray D. N., Echeverri A. Cell-mediated immunity in anergic patients with pulmonary tuberculosis. Am Rev Respir Dis. 1978 Nov;118(5):827–834. doi: 10.1164/arrd.1978.118.5.827. [DOI] [PubMed] [Google Scholar]
  16. Mosmann T. R., Cherwinski H., Bond M. W., Giedlin M. A., Coffman R. L. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol. 1986 Apr 1;136(7):2348–2357. [PubMed] [Google Scholar]
  17. Pancholi P., Mirza A., Bhardwaj N., Steinman R. M. Sequestration from immune CD4+ T cells of mycobacteria growing in human macrophages. Science. 1993 May 14;260(5110):984–986. doi: 10.1126/science.8098550. [DOI] [PubMed] [Google Scholar]
  18. Singh M., Andersen A. B., McCarthy J. E., Rohde M., Schütte H., Sanders E., Timmis K. N. The Mycobacterium tuberculosis 38-kDa antigen: overproduction in Escherichia coli, purification and characterization. Gene. 1992 Aug 1;117(1):53–60. doi: 10.1016/0378-1119(92)90489-c. [DOI] [PubMed] [Google Scholar]
  19. Snider D. E., Jr, Roper W. L. The new tuberculosis. N Engl J Med. 1992 Mar 5;326(10):703–705. doi: 10.1056/NEJM199203053261011. [DOI] [PubMed] [Google Scholar]
  20. Surcel H. M., Troye-Blomberg M., Paulie S., Andersson G., Moreno C., Pasvol G., Ivanyi J. Th1/Th2 profiles in tuberculosis, based on the proliferation and cytokine response of blood lymphocytes to mycobacterial antigens. Immunology. 1994 Feb;81(2):171–176. [PMC free article] [PubMed] [Google Scholar]
  21. Thybo S., Richter C., Wachmann H., Maselle S. Y., Mwakyusa D. H., Mtoni I., Andersen A. B. Humoral response to Mycobacterium tuberculosis-specific antigens in African tuberculosis patients with high prevalence of human immunodeficiency virus infection. Tuber Lung Dis. 1995 Apr;76(2):149–155. doi: 10.1016/0962-8479(95)90558-8. [DOI] [PubMed] [Google Scholar]
  22. Verbon A., Weverling G. J., Kuijper S., Speelman P., Jansen H. M., Kolk A. H. Evaluation of different tests for the serodiagnosis of tuberculosis and the use of likelihood ratios in serology. Am Rev Respir Dis. 1993 Aug;148(2):378–384. doi: 10.1164/ajrccm/148.2.378. [DOI] [PubMed] [Google Scholar]
  23. Vordermeier H. M., Harris D. P., Friscia G., Román E., Surcel H. M., Moreno C., Pasvol G., Ivanyi J. T cell repertoire in tuberculosis: selective anergy to an immunodominant epitope of the 38-kDa antigen in patients with active disease. Eur J Immunol. 1992 Oct;22(10):2631–2637. doi: 10.1002/eji.1830221024. [DOI] [PubMed] [Google Scholar]
  24. Wilkins E. G., Ivanyi J. Potential value of serology for diagnosis of extrapulmonary tuberculosis. Lancet. 1990 Sep 15;336(8716):641–644. doi: 10.1016/0140-6736(90)92144-7. [DOI] [PubMed] [Google Scholar]
  25. Worsaae A., Ljungqvist L., Heron I. Monoclonal antibodies produced in BALB.B10 mice define new antigenic determinants in culture filtrate preparations of Mycobacterium tuberculosis. J Clin Microbiol. 1988 Dec;26(12):2608–2614. doi: 10.1128/jcm.26.12.2608-2614.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Young D., Kent L., Rees A., Lamb J., Ivanyi J. Immunological activity of a 38-kilodalton protein purified from Mycobacterium tuberculosis. Infect Immun. 1986 Oct;54(1):177–183. doi: 10.1128/iai.54.1.177-183.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES