1887

Abstract

Antigens of the outer envelope from serovar (Hond Utrecht IV) were extracted by 50% (v/v) ethanol or by sodium dodecyl sulphate and serological analysis suggested that they were identical. The ‘fraction 4’ extracted by alkali was found to contain glycoproteins of high (retentate) and low (filtrate) molecular weight; the latter behaved like a hapten in serology and in animal immunization experiments. Antibodies were raised in rabbits against this hapten by conjugating it to bovine albumin fraction V. The antiserum was found to react with both the low molecular weight and high molecular weight glycoproteins. This anti-hapten serum contained little or no whole-cell-agglutinating antibodies. The fraction 4 retentate behaved like a complete antigen in serological and immunization studies. Fraction 4 retentate and the outer envelope preparations were serologically related but they were not identical. Chemical studies revealed similarities between the carbohydrate component of the outer envelope obtained by ethanol extraction and fraction 4. The outer envelope extracted by ethanol, fraction 4 and its low and high molecular weight glycoproteins contained arabinose, rhamnose, fucose, xylose, mannose, galactose, glucose, glucosamine and glucuronic acid. Three unidentified peaks were observed in gas-liquid chromatographic analysis of the -trimethylsilyl derivatives of methyl glycosides of all these samples and one of these peaks co-eluted with the -trimethylsilyl derivative of 3--methylmannose.

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1980-03-01
2024-12-05
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References

  1. Adler B., Faine S. 1978a; Serological and protective-antibody responses of rabbits to leptospiral antigens. Journal of Medical Microbiology 11:401–409
    [Google Scholar]
  2. Adler B., Faine S. 1978b; The antibodies involved in the human immune response to leptospiral infection. Journal of Medical Microbiology 11:387–400
    [Google Scholar]
  3. Alton G. G., Jones L. M., Pietz D. E. 1975; Laboratory Techniques in Brucellosis (World Health Organization Monograph Series no. 55). , 2nd. pp. 85–101 Geneva: World Health Organization;
    [Google Scholar]
  4. Auran N. E., Johnson R. C., Ritzi M. 1972; Isolation of the outer sheath of Leptospira and its immunogenic properties in hamsters. Infection and Immunity 5:968–975
    [Google Scholar]
  5. Azuma I., Takeda K., Yamamura Y., Yanagihara Y., Mifuchi I. 1976; Occurrence of 3-O-methylmannose in the polysaccharide of Leptospira biflexa Urawa. Journal of Bacteriology 128:492–494
    [Google Scholar]
  6. Faine S., Adler B., Palit A. 1974a; Chemical, serological and biological properties of serotype- specific antigen in Leptospira. Australian Journal of Experimental Biology and Medical Science 52:311–319
    [Google Scholar]
  7. Faine S., Adler B., Ruta G. 1974b; A mechanism of immunity to leptospirosis. Australian Journal of Experimental Biology and Medical Science 52:301–310
    [Google Scholar]
  8. Fielder R. J., Bishop C. T., Grappel S. F., Blank F. 1970; An immunogenic polysaccharide-protein conjugate. Journal of Immunology 105:265–267
    [Google Scholar]
  9. Hindle E., White P. B. 1934; A specific soluble substance in spirochaetes. Proceedings of the Royal Society 114:523–529
    [Google Scholar]
  10. Kabat E. A., Mayer M. M. 1964 Experimental Immunochemistry, 2nd. (a) pp. 527–528 (b) pp. 505–507 Springfield, Illinois: Charles C. Thomas;
    [Google Scholar]
  11. Lauber K. 1976; Photometric determination of nitrogen. Wet incineration followed by formation of indophenol blue with salicylate/ hypochlorite. Clinica chimica acta 67:107–110
    [Google Scholar]
  12. Likhite V., Sehon A. 1967; 6. Protein-protein conjugation. Methods in Immunology and Immunochemistry 1:150–154
    [Google Scholar]
  13. Lüderitz O., Staub A. M., Westphal O. 1966; Immunochemistry of O and R antigens of Salmonella and related Enterobacteriaceae. Bacteriological Reviews 30:192–255
    [Google Scholar]
  14. Neter E. 1956; Bacterial haemagglutination and haemolysis. Bacteriological Reviews 20:166–188
    [Google Scholar]
  15. Ouchterlony Ö. 1949; Antigen-antibody reactions in gels. Acta pathologica et microbiologica scandinavica 22:507–515
    [Google Scholar]
  16. Palit A., Gulasekharam J. 1973; Genus- specific leptospiral antigen and its possible use in laboratory diagnosis. Journal of Clinical Pathology 26:7–16
    [Google Scholar]
  17. Palit A., Harrison P. M. 1977; Immunochemistry of extracts from Leptospira interrogansserotype hardjo. Journal of General Microbiology 100:249–256
    [Google Scholar]
  18. Palit A., Hamilton R. C., Gulasekharam J. 1974; Further studies onleptospiral genus-specific antigen: its ultrastructure and immunochemistry. Journal of General Microbiology 82:223–236
    [Google Scholar]
  19. Palmer E. L., Martin M. L., Hierholzer J. C., Ziegler D. W. 1971; Nonspecific precipitation of serum proteins by sodium lauryl sulphate in agar diffusion and immunoelectrophoresis. Applied Microbiology 21:903–906
    [Google Scholar]
  20. Singer S. J., Schick A. F. 1961; The properties of specific stains for electron-microscopy prepared by conjugation of antibody molecules with ferritin. Journal of Biophysical and Biochemical Cytology 9:519–537
    [Google Scholar]
  21. Weissbach A., Hurwitz J. 1959; The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. Journal of Biological Chemistry 234:705–709
    [Google Scholar]
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