Abstract
Three hybridomas which secrete antibodies to Mycobacterium avium were obtained by the fusion of p3u1 myeloma cells with spleen cells of mice immunized with M. avium culture sonicate. The reactivity of these monoclonal antibodies was determined in 16 species of mycobacteria by an enzyme-linked immunosorbent assay. An antibody, designated Avi-3, reacted only with M. avium and not with the reference strains of the other 15 species of mycobacteria tested, including M. intracellulare, M. paratuberculosis, and M. lepraemurium. Specificity of the antibody was confirmed by assay, using a specific DNA probe of M. avium complex in 29 M. avium complex isolates. An antigen was purified from M. avium culture sonicate on a monoclonal antibody Avi-3-coupled affinity column. The purified antigen gave a single band (molecular size, about 27 kilodaltons) upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigen Avi-3 showed strong skin test activity in guinea pig preimmunized with heat-killed M. avium but not in those sensitized with heat-killed M. intracellulare or M. bovis BCG. Purified protein derivative elicited positive skin reactions in all the immunized guinea pigs. When heat-killed M. avium was used as the immunogen, strong lymphoproliferative responses were observed in cultures stimulated with the antigen Avi-3. These results suggest that M. avium-specific antigen Avi-3 may facilitate the diagnosis of mycobacterial infections.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Baess I. Deoxyribonucleic acid relatedness among species of slowly-growing mycobacteria. Acta Pathol Microbiol Scand B. 1979 Aug;87(4):221–226. doi: 10.1111/j.1699-0463.1979.tb02430.x. [DOI] [PubMed] [Google Scholar]
- Baess I. Deoxyribonucleic acid relationships between different serovars of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum. Acta Pathol Microbiol Immunol Scand B. 1983 Jun;91(3):201–203. doi: 10.1111/j.1699-0463.1983.tb00033.x. [DOI] [PubMed] [Google Scholar]
- Bloom B. R., Godal T. Selective primary health care: strategies for control of disease in the developing world. V. Leprosy. Rev Infect Dis. 1983 Jul-Aug;5(4):765–780. doi: 10.1093/clinids/5.4.765. [DOI] [PubMed] [Google Scholar]
- Brennan P. J., Goren M. B. Structural studies on the type-specific antigens and lipids of the mycobacterium avium. Mycobacterium intracellulare. Mycobacterium scrofulaceum serocomplex. Mycobacterium intracellulare serotype 9. J Biol Chem. 1979 May 25;254(10):4205–4211. [PubMed] [Google Scholar]
- Britton W. J., Hellqvist L., Basten A., Raison R. L. Mycobacterium leprae antigens involved in human immune responses. I. Identification of four antigens by monoclonal antibodies. J Immunol. 1985 Dec;135(6):4171–4177. [PubMed] [Google Scholar]
- Buchanan T. M., Nomaguchi H., Anderson D. C., Young R. A., Gillis T. P., Britton W. J., Ivanyi J., Kolk A. H., Closs O., Bloom B. R. Characterization of antibody-reactive epitopes on the 65-kilodalton protein of Mycobacterium leprae. Infect Immun. 1987 Apr;55(4):1000–1003. doi: 10.1128/iai.55.4.1000-1003.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Drake T. A., Hindler J. A., Berlin O. G., Bruckner D. A. Rapid identification of Mycobacterium avium complex in culture using DNA probes. J Clin Microbiol. 1987 Aug;25(8):1442–1445. doi: 10.1128/jcm.25.8.1442-1445.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gillis T. P., Buchanan T. M. Production and partial characterization of monoclonal antibodies to Mycobacterium leprae. Infect Immun. 1982 Jul;37(1):172–178. doi: 10.1128/iai.37.1.172-178.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hawkins C. C., Gold J. W., Whimbey E., Kiehn T. E., Brannon P., Cammarata R., Brown A. E., Armstrong D. Mycobacterium avium complex infections in patients with the acquired immunodeficiency syndrome. Ann Intern Med. 1986 Aug;105(2):184–188. doi: 10.7326/0003-4819-105-2-184. [DOI] [PubMed] [Google Scholar]
- 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]
- Ivanyi J., Sinha S., Aston R., Cussell D., Keen M., Sengupta U. Definition of species specific and cross-reactive antigenic determinants of Mycobacterium leprae using monoclonal antibodies. Clin Exp Immunol. 1983 Jun;52(3):528–536. [PMC free article] [PubMed] [Google Scholar]
- Kiehn T. E., Edwards F. F. Rapid identification using a specific DNA probe of Mycobacterium avium complex from patients with acquired immunodeficiency syndrome. J Clin Microbiol. 1987 Aug;25(8):1551–1552. doi: 10.1128/jcm.25.8.1551-1552.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kolk A. H., Ho M. L., Klatser P. R., Eggelte T. A., Kuijper S., de Jonge S., van Leeuwen J. Production and characterization of monoclonal antibodies to Mycobacterium tuberculosis, M. bovis (BCG) and M. leprae. Clin Exp Immunol. 1984 Dec;58(3):511–521. [PMC free article] [PubMed] [Google Scholar]
- Köhler G., Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. doi: 10.1002/eji.1830060713. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Minden P., Kelleher P. J., Freed J. H., Nielsen L. D., Brennan P. J., McPheron L., McClatchy J. K. Immunological evaluation of a component isolated from Mycobacterium bovis BCG with a monoclonal antibody to M. bovis BCG. Infect Immun. 1984 Nov;46(2):519–525. doi: 10.1128/iai.46.2.519-525.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morris J. A., Thorns C. J., Woolley J. The identification of antigenic determinants on Mycobacterium bovis using monoclonal antibodies. J Gen Microbiol. 1985 Jul;131(7):1825–1831. doi: 10.1099/00221287-131-7-1825. [DOI] [PubMed] [Google Scholar]
- Schaefer W. B. Serologic identification and classification of the atypical mycobacteria by their agglutination. Am Rev Respir Dis. 1965 Dec;92(6):85–93. doi: 10.1164/arrd.1965.92.6P2.85. [DOI] [PubMed] [Google Scholar]
- Schaefer W. B. Type-specificity of atypical mycobacteria in agglutination and antibody absorption tests. Am Rev Respir Dis. 1967 Dec;96(6):1165–1168. doi: 10.1164/arrd.1967.96.6.1165. [DOI] [PubMed] [Google Scholar]
- Stanford J. L., Grange J. M. The meaning and structure of species as applied to mycobacteria. Tubercle. 1974 Jun;55(2):143–152. doi: 10.1016/0041-3879(74)90008-7. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wolinsky E. Nontuberculous mycobacteria and associated diseases. Am Rev Respir Dis. 1979 Jan;119(1):107–159. doi: 10.1164/arrd.1979.119.1.107. [DOI] [PubMed] [Google Scholar]
- Zakowski P., Fligiel S., Berlin G. W., Johnson L., Jr Disseminated Mycobacterium avium-intracellulare infection in homosexual men dying of acquired immunodeficiency. JAMA. 1982 Dec 10;248(22):2980–2982. doi: 10.1001/jama.1982.03330220024029. [DOI] [PubMed] [Google Scholar]