1887

Abstract

Summary: To gain insight into the pathogenesis of tuberculosis, a molecular definition of the tubercle bacillus cell envelope, which is involved in the early stages of the infection, is required. The cell-surface-exposed material of the pathogen was isolated by mechanical means and chemically analysed. It was shown by scanning electron microscopy that the method used for extracting the surface-covering material preserves the integrity of the bacilli. Surprisingly, in view of the current opinion, only small amounts of lipids (1-6%) were present. Polysaccharides and proteins were the main components of the material. The polysaccharides were neutral and lipid-free D-glucan, D-arabino-D-mannan and D-mannan, which were eluted from gel-filtration columns in positions corresponding to molecular masses of 120, 13 and 4 kDa, respectively. Based on NMR spectroscopy and conventional chemical analyses, the major structural motifs of the purified polysaccharides were established as being identical to those of the polysaccharides we previously isolated from the culture filtrate of the tubercle bacillus. Immunocytochemical studies showed that these compounds were not only surface-located but were also present in the inner capsular compartment. The major protein constituents exhibited the same mobilities on SDS-PAGE as those of the culture filtrate of the tubercle bacillus and readily reacted with the monoclonal antibodies directed against these molecules. These proteins included the 19 and 38 kDa lipoproteins, the 30/31 kDa fibronectin-binding proteins and the 40 kDa L-alanine dehydrogenase. These findings suggest that the culture filtrate material represents part of the capsule which, in an context, could contribute to the electron transparent zone surrounding the tubercle bacillus. The 24 kDa (MPB/T64) protein was found to be a secreted protein, as it was detected almost exclusively in the culture filtrate. Taken together, the data give a new insight into the surface-exposed compounds of the tubercle bacillus and may explain part of the nature and limitation of the host immunity towards the pathogen.

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-7-1609
1995-07-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/7/mic-141-7-1609.html?itemId=/content/journal/micro/10.1099/13500872-141-7-1609&mimeType=html&fmt=ahah

References

  1. Abou-Zeid C., Ratliff T.L, Harboe M., Bennedsen J., Rook G.A.W. 1988; Characterization of fibronectin-binding antigens released by Mycobacterium tuberculosis and Mycobacterium bovis BCG.. Infect Immun 56:3046–3051
    [Google Scholar]
  2. Amar-Nacasch C., Vilkas E. 1970; Etude des parois de Mycobacterium tuberculosis. II. Mise en evidence d’un mycolate d’arabinobiose et d’un glucane dans les parois de M. tuberculosis H37Ra.. Hull Soc Chim Biol 52:145–151
    [Google Scholar]
  3. Andersen A.B., Yuan Z.L., Haslov K., Vergmann B., Benned-sen J. 1986; Interspecies reactivity of five monoclonal antibodies to Mycobacterium tuberculosis as examined by immunoblotting and enzyme-linked immunosorbent assay.. Infect Immun 23:446–451
    [Google Scholar]
  4. Andersen A.B., Ljungqvist L., Haslov K., Bentzon M.W. 1991; MPB 64 possesses ‘tuberculosis-complex’-specific B- and T-cell epitopes.. Scand J Immunol 34:365–372
    [Google Scholar]
  5. Andersen A.B., Andersen P., Ljungqvist L. 1992; Structure and function of a 40,000-molecular-weight protein antigen of Mycobacterium tuberculosis.. Infect Immun 60:2317–2323
    [Google Scholar]
  6. Andersen P., Askgaard D., Ljungqvist L., Bennedsen J., Heron I. 1991; Proteins released from Mycobacterium tuberculosis during growth.. Infect Immun 59:1905–1910
    [Google Scholar]
  7. Antoine A.D., Tepper B.S. 1969; Characterization of glyco-gens from mycobacteria.. Arch Biochem Biophys 134:207–213
    [Google Scholar]
  8. Armstrong J.A., D’ArcyHart P.D. 1971; Response of cultured macrophages to Mycobacterium tuberculosis with observation on fusion of lysosomes with phagosomes. J Exp Med 134:713–740
    [Google Scholar]
  9. Armstrong J.A., D’Arcy Hart P.D. 1975; Phagosome-lysosome interactions in cultured macrophages infected with virulent tubercle bacilli. Reversal of the usual non-fusion pattern and observation on bacterial survival.. J Exp Med 142:1–16
    [Google Scholar]
  10. Bloom B.R., Murray J.L. 1992; Turberculosis: commentary on a reemergent killer.. Science 257:1055–1064
    [Google Scholar]
  11. Boddingius J., Dijkman H.P. 1989; Immunogold labelling method for Mycobacterium leprae-specific phenolic glycolipid in glutaraldehyde-osmium-fixed and araldite-embedded leprosy lesion.. J Histochem Cytochem 37:455–462
    [Google Scholar]
  12. Bradbury J.H., Jenkins G.A. 1984; Determination of the structures of trisaccharides by 13C-NMR spectroscopy.. Carbohydr Res 126:125–156
    [Google Scholar]
  13. Buttiaux R., Beerens H. 1966; Mycobacteries.. In Manuel de Techniques Bacteriologiques, 2nd edn. pp. 477–547 Buttiaux R., Beerens H., Tacquet A. Edited by Paris: Flammarion;
    [Google Scholar]
  14. Chatterjee D., Lowell K., Rivoire B., McNeil M.R., Brennan P. 1992; Lipoarabinomannan of Mycobacterium tuberculosis-, capping with mannosyl residues in some strains.. J Biol Chem 267:6234–6239
    [Google Scholar]
  15. Ciucanu I., Kerek F. 1984; A simple and rapid method for permethylation of carbohydrates.. Carbohydr Res 131:209–217
    [Google Scholar]
  16. Daffé M., Lacave C., Lanéelle M.-A., Lanéelle G. 1987; Structure of the major triglycosyl phenol-phthiocerol of Myco-bacterium tuberculosis (strain Canetti).. Eur J Biochem 167:155–160
    [Google Scholar]
  17. Daffé M., Brennan P.J., McNeil M. 1990; 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 265:6734–6743
    [Google Scholar]
  18. Daffé M., Cho S.-N., Chatterjee D., Brennan P.J. 1991a; Chemical synthesis and seroreactivity of a neoantigen containing the oligosaccharide hapten of the Mycobacterium tuberculosis-specific phenolic glycolipid.. J Infect Dis 163:161–168
    [Google Scholar]
  19. Daffé M., McNeil M., Brennan P.J. 1991b; Novel type-specific lipooligosaccharides from Mycobacterium tuberculosis.. Biochemistry 30:378–388
    [Google Scholar]
  20. Dische Z. 1962; Color reaction of hexoses.. Methods Carbohydr Chem 1:488–494
    [Google Scholar]
  21. Draper P. 1982; The anatomy of mycobacteria.. In The Biology of the Mycobacteria 1 pp. 9–52 Ratledge C.C., Stanford J.L. Edited by London: Academic Press;
    [Google Scholar]
  22. Draper P., Rees R.J.W. 1970; Electron transparent zone of mycobacteria may be a defence mechanism.. Nature 228:860–861
    [Google Scholar]
  23. Draper P., Rees R.J.W. 1973; The nature of the electron- transparent zone that surrounds Mycobacterium lepraemurium inside host cells.. J Gen Microbiol 77:79–87
    [Google Scholar]
  24. Ellner J.J., Daniel T.M. 1979; Immunosuppression by mycobacterial arabinomannan.. Clin Exp Immunol 35:250–257
    [Google Scholar]
  25. Espitia C., Elinos M., Hernandez-Pando R., Mancilla R.A. 1992; Phosphate starvation enhances expression of the immuno-dominant 38-kilodalton protein antigen of Mycobacterium tuberculosis-. demonstration by immunogold electron microscopy.. Inject Immun 60:2998–3001
    [Google Scholar]
  26. Fournié J.J., Mullins R.J., Basten A. 1991; Isolation and structural characteristics of a monoclonal antibody-defined cross-reactive phospholipid antigen from Mycobacterium tuberculosis and Mycobacterium leprae.. J Biol Chem 266:1211–1219
    [Google Scholar]
  27. Gaylord H., Brennan P.J., Young D.B., Buchanan T.M. 1987; Most Mycobacterium leprae carbohydrate-reactive monoclonal antibodies are directed to lipoarabinomannan.. Infect Immun 55:2860–2863
    [Google Scholar]
  28. Gerwig G.J., Kamerling J.B., Vliegenthart J.F.G. 1978; Determination of the D and L configuration of neutral monosaccharides by high-resolution capillary GLC.. Carbohydr Res 62:349–357
    [Google Scholar]
  29. Goren M.B., Brennan P.J. 1979; Mycobacterial lipids.. In Tuberculosis pp. 69–193 Youmans G.P. Edited by Philadelphia: W.B.Saunders;
    [Google Scholar]
  30. Hanks J.H. 1961; Significance of capsular components of Mycobacterium leprae and other mycobacteria.. Int J Eepr 29:74–83
    [Google Scholar]
  31. Hunter S.W., Brennan P.J. 1983; Further specific phenolic glycolipid antigens and a related diacyl phthiocerol from Myco-bacterium leprae.. J Biol Chem 258:7556–7562
    [Google Scholar]
  32. Kochi A. 1991; The global tuberculosis situation and the new control strategy of the World Health Organization.. Tubercle 72:1–6
    [Google Scholar]
  33. Koul A.K., Gastambide-Odier M. 1977; Microanalyse rapide de dimycocerosate de phtiocerol, de mycosides et de glycerides dans les extraits a l’ether de petrole de Mycobacterium kansasii et du BCG, souche Pasteur.. Biochimie 59:535–538
    [Google Scholar]
  34. Kreis B. 1966; La mesure de la resistance.. In Resistance et Survivance du Bacille Tuberculeux aux Medicaments Antibacillaires pp. 45–73 Kreis B. Edited by Paris: Masson et Cie ;
    [Google Scholar]
  35. Lemassu A., Daffé M. 1994; Structural features of the exocellular polysaccharides of Mycobacterium tuberculosis.. Biochem J 297:351–357
    [Google Scholar]
  36. Ljungqvist L., Worsaae A., Heron I. 1988; Antibody responses against Mycobacterium tuberculosis in 11 strains of inbred mice: novel monoclonal antibody specificities generated by fusions, using spleens from BALB. B10 and CBA/J mice.. Infect Immun 56:1994–1998
    [Google Scholar]
  37. Merckx J.J., Brown A.L., Karlson A.G. 1964; An electron- microscopic study of experimental infection with acid-fast bacilli.. Am Rev Respir Dis 89:485–496
    [Google Scholar]
  38. Misaki A., Yukawa S. 1966; Studies on cell walls of mycobacteria. II. Constitution of polysaccharides from BCG cell walls.. J Biochem 59:511–520
    [Google Scholar]
  39. Moreno C., Mehlert A., Lamb J. 1988; The inhibitory effects of mycobacterial lipoarabinomannan and polysaccharides upon polyclonal and monoclonal human T cell proliferation.. Clin Exp Immunol 74:206–210
    [Google Scholar]
  40. Patel A.M., Abraham E.W. 1989; Pulmonary tuberculosis.. In The Biology of the Mycobacteria 3 pp. 179–244 Ratledge C., Stanford J., Grange J.M. Edited by London: Academic Press;
    [Google Scholar]
  41. Picard B., Frehel C., Rastogi N. 1984; Cytochemical characterization of mycobacterial outer surfaces.. Acta Eepr 95:227–235
    [Google Scholar]
  42. Rambukkana A., Das P.K.A., Chand A., Baas J.G., Groothuis D.G., Kolk A.H.J. 1991; Subcellular distribution of monoclonal antibody defined epitopes on immunodominant Mycobacterium tuberculosis proteins in the 30-kDa region: identification and localization of 29/33-kDa doublet proteins on mycobacterial cell wall.. Scand J Immunol 33:763–775
    [Google Scholar]
  43. Sauton B. 1912; Sur la nutrition minerale du bacille tuberculeux.. C.R Acad Sci Ser III Sci Vie 155:860–863
    [Google Scholar]
  44. Schlesinger L.S. 1993; Macrophage phagocytosis of virulent but not attenuated strains of Mycobacterium tuberculosis is mediated by mannose receptors in addition to complement receptors.. J Immunol 150:2920–2930
    [Google Scholar]
  45. Schlesinger L.S., Hull S.R., Kaufman T.M. 1994; Binding of the terminal mannosyl units of lipoarabinomannan from a virulent strain of Mycobacterium tuberculosis to human macrophages.. J Immunol 152:4070–4079
    [Google Scholar]
  46. Stewart T.S., Ballou C.E. 1968; A comparison of yeast mannans and phosphomannans by acetolysis.. Biochemistry 7:1855–1863
    [Google Scholar]
  47. Venisse A., Berjeaud J.M., Chaurand P., Gilleron M., Puzo G. 1993; Structural features of lipoarabinomannan from Myco-bacterium bovis BCG.. J Biol Chem 268:12401–12411
    [Google Scholar]
  48. Wiker H.G., Harboe M. 1992; The antigen 85 complex: a major secretion product of Mycobacterium tuberculosis.. Microbiol Rev 56:648–661
    [Google Scholar]
  49. Zang L.-H., Howseman A.M., Shulman R.G. 1991; Assignment of 1H chemical shifts of glycogen.. Carbohydr Res 220:1–9
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-7-1609
Loading
/content/journal/micro/10.1099/13500872-141-7-1609
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error