Skip to main content
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1992 Jul;89(1):115–119. doi: 10.1111/j.1365-2249.1992.tb06888.x

Monoclonal antibody epitopes of mycobacterial 65-kD heat-shock protein defined by epitope scanning.

A H Hajeer 1, J Worthington 1, K Morgan 1, R M Bernstein 1
PMCID: PMC1554412  PMID: 1378362

Abstract

The binding sites for MoAbs to the 65-kD heat-shock protein (hsp65) of mycobacteria have been investigated by epitope scanning. Five hundred and twenty-six 8-mer peptides representing the complete sequence of Mycobacterium tuberculosis hsp65 were synthesised in duplicate using the Epitope Scanning Kit (CRB Ltd.). The epitopes of six MoAbs raised to the hsp65 of M. tuberculosis or M. leprae were investigated. We have identified the epitope of a new MoAb (DC16); this epitope is continuous, hydrophilic in nature and 11 amino acids long. We have also confirmed the location of the epitopes of three MoAbs (IIH9, ML30 and IIC8). Thus the epitope scanning technique has proved suitable for the detection of continuous epitopes of hsp65.

Full text

PDF
115

Selected References

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

  1. Anderson D. C., Barry M. E., Buchanan T. M. Exact definition of species-specific and cross-reactive epitopes of the 65-kilodalton protein of Mycobacterium leprae using synthetic peptides. J Immunol. 1988 Jul 15;141(2):607–613. [PubMed] [Google Scholar]
  2. Bahr G. M., Rook G. A., al-Saffar M., Van Embden J., Stanford J. L., Behbehani K. Antibody levels to mycobacteria in relation to HLA type: evidence for non-HLA-linked high levels of antibody to the 65 kD heat shock protein of M. bovis in rheumatoid arthritis. Clin Exp Immunol. 1988 Nov;74(2):211–215. [PMC free article] [PubMed] [Google Scholar]
  3. Brand S. R., Worthington J., McIntosh D. P., Bernstein R. M. Antibody to a 63 kilodalton insect protein in ankylosing spondylitis. Ann Rheum Dis. 1992 Mar;51(3):334–339. doi: 10.1136/ard.51.3.334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Cohen I. R., Holoshitz J., van Eden W., Frenkel A. T lymphocyte clones illuminate pathogenesis and affect therapy of experimental arthritis. Arthritis Rheum. 1985 Aug;28(8):841–845. doi: 10.1002/art.1780280802. [DOI] [PubMed] [Google Scholar]
  6. Emmrich F., Thole J., van Embden J., Kaufmann S. H. A recombinant 64 kilodalton protein of Mycobacterium bovis bacillus Calmette-Guerin specifically stimulates human T4 clones reactive to mycobacterial antigens. J Exp Med. 1986 Apr 1;163(4):1024–1029. doi: 10.1084/jem.163.4.1024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Geysen H. M., Meloen R. H., Barteling S. J. Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proc Natl Acad Sci U S A. 1984 Jul;81(13):3998–4002. doi: 10.1073/pnas.81.13.3998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Holoshitz J., Koning F., Coligan J. E., De Bruyn J., Strober S. Isolation of CD4- CD8- mycobacteria-reactive T lymphocyte clones from rheumatoid arthritis synovial fluid. Nature. 1989 May 18;339(6221):226–229. doi: 10.1038/339226a0. [DOI] [PubMed] [Google Scholar]
  9. Jindal S., Dudani A. K., Singh B., Harley C. B., Gupta R. S. Primary structure of a human mitochondrial protein homologous to the bacterial and plant chaperonins and to the 65-kilodalton mycobacterial antigen. Mol Cell Biol. 1989 May;9(5):2279–2283. doi: 10.1128/mcb.9.5.2279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kleinau S., Söderström K., Kiessling R., Klareskog L. A monoclonal antibody to the mycobacterial 65 kDa heat shock protein (ML 30) binds to cells in normal and arthritic joints of rats. Scand J Immunol. 1991 Feb;33(2):195–202. doi: 10.1111/j.1365-3083.1991.tb03749.x. [DOI] [PubMed] [Google Scholar]
  11. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Rambukkana A., Yong S., Das P. K. Identification of a novel B-cell epitope of restricted specificity on the hsp 65-kDa protein of Mycobacterium tuberculosis. FEMS Microbiol Immunol. 1991 Feb;3(1):39–45. doi: 10.1111/j.1574-6968.1991.tb04161.x. [DOI] [PubMed] [Google Scholar]
  14. Shinnick T. M. The 65-kilodalton antigen of Mycobacterium tuberculosis. J Bacteriol. 1987 Mar;169(3):1080–1088. doi: 10.1128/jb.169.3.1080-1088.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Stark S. E., Caton A. J. Antibodies that are specific for a single amino acid interchange in a protein epitope use structurally distinct variable regions. J Exp Med. 1991 Sep 1;174(3):613–624. doi: 10.1084/jem.174.3.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Worthington J., Brass A., Morgan K. Identification of antibody epitopes within the CB-11 peptide of type II collagen. I: Detection of antibody binding sites by epitope scanning. Autoimmunity. 1991;10(3):201–207. doi: 10.3109/08916939109001890. [DOI] [PubMed] [Google Scholar]
  17. van Eden W., Thole J. E., van der Zee R., Noordzij A., van Embden J. D., Hensen E. J., Cohen I. R. Cloning of the mycobacterial epitope recognized by T lymphocytes in adjuvant arthritis. Nature. 1988 Jan 14;331(6152):171–173. doi: 10.1038/331171a0. [DOI] [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES