1887

Abstract

Summary

A 66-kDa protein (TB66) was purified from culture filtrate (CF) and cell sonicate (CS) of HRv by immobilised metal affinity chromatography (IMAC) on a Ni-nitrilotriacetic acid (NTA) column. TB66 was found to be a fibronectin-binding protein as determined by ELISA and could be purified by affinity chromatography with fibronectin-Sepharose. A similar 66-kDa protein could be isolated also from BCG, and HRa by IMAC, but not from any other mycobacteria. The NH-terminal amino-acid sequence of TB66 from HRv and was identical and showed 85% homology with the N-terminal sequence of bovine serum albumin (BSA). A monoclonal antibody (MAb) OD4AG3 recognised a heat-stable and trypsin-sensitive epitope near the C-terminal end of TB66. This MAb also recognised the 66-kDa protein isolated from the other members of the complex. In tests of immunogenicity, TB66 elicited a delayed type hypersensitivity reaction in guinea-pigs immunised with either TB66 or with H37Rv. TB66 also elicited an antibody response in immunised guinea-pigs and stimulated murine macrophages to produce tumour necrosis factor.

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1994-12-01
2022-01-16
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References

  1. Murray CJ, Styblo K, Rouillon A. Tuberculosis in developing countries: burden, intervention and cost. Bull Int Union Tuberc Lung Dis 1990; 65:6–24
    [Google Scholar]
  2. Daniel TM, Debanne SM. The serodiagnosis of tuberculosis and other mycobacterial diseases by enzyme-linked immunosorbent assay. Am Rev Respir Dis 1987; 135:1137–1151
    [Google Scholar]
  3. Daniel TM, Janicki BW. Mycobacterial antigens: a review of their isolation, chemistry, and immunological properties. Microbiol Rev 1978; 42:84–113
    [Google Scholar]
  4. Stanford JL. Immunologically important constituents of mycobacteria: antigens. In Ratledge C, Stanford J. (eds) The biology of mycobacteria, vol. 2. Immunological and environmental aspects London: Academic Press Inc.; 198385–127
    [Google Scholar]
  5. Abou-Zeid C, Smith I, Grange JM, Ratliff TL, Steele J, Rook GAW. The secreted antigens of Mycobacterium tuberculosis and their relationship to those recognized by the available antibodies. J Gen Microbiol 1988; 134:531–538
    [Google Scholar]
  6. Abou-Zeid C, Ratliff TL, Wiker HG, Harboe M, Bennedsen J, Rook GAW. Characterization of fibronectin-binding antigens released by Mycobacterium tuberculosis and Mycobacterium bovis BCG. Infect Immun 1988; 56:3046–3051
    [Google Scholar]
  7. Hynes RO, Yamada KM. Fibronectins: multifunctional modular glycoproteins. J Cell Biol 1982; 95:369–377
    [Google Scholar]
  8. Ratliff TL, McGarr JA, Abou-Zeid GAW. et al. Attachment of mycobacteria to fibronectin-coated surfaces. J Gen Microbiol 1988; 134:1307–1313
    [Google Scholar]
  9. Valone SE, Rich EA, Wallis RS, Ellner JJ. Expression of tumor necrosis factor in vitro by human mononuclear phagocytes stimulated with whole Mycobacterium bovis BCG and mycobacterial antigens. Infect Immun 1988; 56:3313–3315
    [Google Scholar]
  10. Wallis RS, Amir-Tahmasseb M, Ellner JJ. Induction of interleukin 1 and tumor necrosis factor by mycobacterial proteins: the monocyte western blot. Proc Natl Acad Sci USA 1990; 87:3348–3352
    [Google Scholar]
  11. Deshpande RG, Khan MB, Bhat DA, Navalkar RG. Purification and partial characterisation of novel 66-kDa seroreactive protein of Mycobacterium tuberculosis H37Rv. J Med Microbiol 1994; 41:173–178
    [Google Scholar]
  12. Hochuli E, Dobeli E, Schacher A. New metal chelate adsorbent selective for proteins and peptides containing neighbouring histidine residues. J Chromatogr 1987; 411:177–184
    [Google Scholar]
  13. Vuento M. Purification of fibronectin from human plasma by affinity chromatography under non-denaturing conditions. Biochem J 1979; 183:331–337
    [Google Scholar]
  14. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76:4350–4354
    [Google Scholar]
  15. Deshpande RG, Khan MB, Navalkar RG. Immunoreactivity of a mammalian liver component with leprosy sera. Int Arch Allergy Immunol 1992; 97:345–349
    [Google Scholar]
  16. Flick DA, Gifford GE. Comparison of in vitro cell cytotoxic assays for tumor necrosis factor. J Immunol Methods 1984; 68:167–175
    [Google Scholar]
  17. McGavin MJ, Raucci G, Gurusiddappa S, Hook M. Fibronectin binding determinants of the Staphylococcus aureus fibronectin receptor. J Biol Chem 1991; 266:8343–8347
    [Google Scholar]
  18. Wallis RS, Paranjape R, Phillips M. Identification by twodimensional gel electrophoresis of a 58-kilodalton tumor necrosis factor-inducing protein of Mycobacterium tuberculosis. Infect Immun 1993; 61:627–632
    [Google Scholar]
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