1887

Abstract

A low- factor which induces gonoccocal resistance to complement-mediated serum killing has been partially purified from lysates of mixed red and buffy coat cells from human blood. The lysates were dialysed against Tris buffer for 24 h at 25 °C with the diffusate being continuously recycled through a column of QAE-Sephadex A25. After elution in an NaCl gradient, the active fractions were both desalted and further purified on Sephadex G10. A second fractionation on QAE-Sephadex A25 and desalting with Sephadex G10 preceded further purification by repeated high-pressure liquid chromatography (HPLC) using a DEAE anion exchange column and desalting with Sephadex G10. Less than 500 μg of material showing one peak in HPLC was obtained from 1 litre of blood. After NMR had indicated the possible presence of pyrimidine nucleotide, carbohydrate and -acetyl groups, nanogram quantities of a commercial preparation of cytidine 5′-monophospho--acetylneuraminic acid (CMP-NANA) were shown to induce gonococci to serum resistance. The synthetic CMP-NANA also co-eluted with the preparation from blood cells in HPLC, and the two materials were indistinguishable in their patterns of acid and heat lability. Furthermore, the resistance-inducing activity of both materials was inhibited by cytidine monophosphate, which is known to inhibit sialylation reactions by CMP-NANA. It appears therefore that the resistance-inducing factor is CMP-NANA or a closely related compound. If the factor is CMP-NANA, biological activities indicated that the cell lysate from 1 litre of blood contained about 40 μg, and the most purified preparation contained only about 1%. With this minute amount in a mixture, the presence of CMP-NANA or a closely related analogue could not be established unequivocally by NMR.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-12-3295
1988-12-01
2021-05-14
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/12/mic-134-12-3295.html?itemId=/content/journal/micro/10.1099/00221287-134-12-3295&mimeType=html&fmt=ahah

References

  1. Apicella M.A., Shero M., Jarvis G.A., Griffiss J.M., Mandrbll R.E., Schneider H. 1987; Phenotypic variation in epitope expression of the Neisseria gonorrhoeae lipooligosaccharide.. Infection and Immunity 55:1755–1761
    [Google Scholar]
  2. Apicella M.A., Wbsternick M.A.J., Morse S.A., Schneider H., Ricb P.A., Griffiss J.M. 1986; Bactericidal antibody response of normal human serum to the lipooligosaccharide of Neisseria gonorrhoeae. . Journal of Infectious Diseases 153:520–525
    [Google Scholar]
  3. Bbrnacki R.J. 1975; Regulation of rat-liver glycoprotein; jy-acetyineuraminic acid transferase activity by pyrimidine nucleotides.. European Journal of Biochemistry 58:477–481
    [Google Scholar]
  4. Bernt E., Bbromeybr H.U. 1974 Methods of Enzymatic Analysis, 2nd edn.. pp 1643–1647 Weinheim: Verlag Chemie;
    [Google Scholar]
  5. Brackmann T.R., Geldmbyer S., Jahn R., Soling H.D. 1983; Radioenzymatic determination of CMP-N-acetylsialic acid and free N-acetyl- sialic acid in biological material.. Analytical Biochemistry 130:369–375
    [Google Scholar]
  6. Brown M.R.W., Williams P. 1985; The influence of environment on envelope properties affecting survival of bacteria in infection.. Annual Review of Microbiology 39:527–556
    [Google Scholar]
  7. Carey D.J., Hirschbbrg C.D. 1979; Metabolism of Af-acetylneuraminic acid in mammals: isolation and characterisation of CMP-lV-acetylneuraminic acid.. Biochemistry 18:2086–2092
    [Google Scholar]
  8. Ferwerda W., Blok C.M., Van-Rinsum J. 1983; Synthesis of N-acetylneuraminic acid and CMP-Y-acetylneuraminic acid in the rat liver cell.. Biochemical Journal 216:87–92
    [Google Scholar]
  9. Glynn A.A., Ward M.E. 1970; Nature and heterogeneity of the antigens of Neisseria gonorrhoeae involved in serum bactericidal reaction.. Infection and Immunity 2:162–168
    [Google Scholar]
  10. Gottschalk A. 1972 Glycoproteins. Their Composition, Structure and Function Amsterdam: Elsevier;
    [Google Scholar]
  11. Griffiss J.M., O’Brien J.P., Yamasaki R., Williams G.D., Ride P.A., Schneider H. 1987; Physical heterogeneity of neisserial lipo- oligosaccharides reflects oligosaccharides that differ in apparent molecular weight, chemical composition and antigenic expression.. Infection and Immunity 55:1792–1800
    [Google Scholar]
  12. Guymon L.F., Lbb T.J., Walstad D., Schmayer A., Sparling P.F. 1978; Altered outer membrane components in serum-sensitive and serum- resistant strains of Neisseria gonorrhoeae. . In Immunobiology of Neisseria gonorrhoeae pp 139–141 Brooks G. F., Gotschlich E. C., Holmes K. K., Sawyer W. D., Young F. E. Edited by Washington, DC: American Society for Microbiology;
    [Google Scholar]
  13. Guymon L.F., Esser M., Shafer W.M. 1982; Pyocin-resistant lipopoiysaccharide mutants of Neisseria gonorrhoeae: alterations in sensitivity to normal human serum and polymyxin B.. Infection and Immunity 36:541–547
    [Google Scholar]
  14. Igarashi M., Komiva Y., Kurokawa M. 1985; CMP-sialk acid, the sde sialosyl donor, is intra- axionally transported.. FEBS Letters 192:239–242
    [Google Scholar]
  15. Martin P.M.V., Patel P.V., Parsons N.J., Smith H. 1981; Induction of phenotypically determined resistance of Neisseria gonorrhoeae to human serum by factors in human serum.. Journal of General Microbiology 127:213–217
    [Google Scholar]
  16. Martin P.M.V., Patel P.V., Parsons N.J., Smith H. 1982; Induction in gonococci of phenotypic resistance to killing by human serum by genital secretions.. British Journal of Venereal Disease 58:363–365
    [Google Scholar]
  17. Martin P.M.V., Patel P.V., Clay J.C., Parsons N.J., Smith H. 1984; Induction of serum resistance of Neisseria gonorrhoeae by human serum: a clinical survey of patients with gonorrhoea.. British Journal of Venereal Disease 60:151–153
    [Google Scholar]
  18. Parsons N.J., Patbl P.V., Martin P.M.V., Goldner M., Smith H. 1985; Gonococci in vitro and in vivo: studies of the host and bacterial determinants of gonococcal resistance to killing by human serum and by phagocytes.. In The Pathogenic Neisseriae pp 487–494 Schoolnik G. K., Brookes G. F., Falkow S., Frasch C. F., Knapp J. S., Mccutchan J. A., Morse S. A. Edited by Washington, DC: American Society for Microbiology;
    [Google Scholar]
  19. Parsons N.J., Patel P.V., Tan E.L., Andrade J.R.C., Nairn C.A., Goldner M., Cole J.A., Smith H. 1988; Cytidine 5´-monophospho-.V- acetyl neuraminic acid and a low molecular weight factor from human blood cells induce lipopolysac- charide alteration in gonococci when conferring resistance to killing by human serum.. Microbial Pathogenesis in the Press
    [Google Scholar]
  20. Patel P.V., Martin P.M.V., Goldner M., Parsons N.J., Smith H. 1984a; Red blood cells, a source of factors which induce Neisseria gonorrhoeae to resistance to complement-mediated killing by human sera.. Journal of General Microbiology 130:2767–2700
    [Google Scholar]
  21. Patel P.V., Vealb D., Fox J.E., Martin P.M.V., Parsons N.J., Smith H. 1984b; Fractionation of guinea pig serum for an inducer of gonococcal resistance to killing by human serum: active fractions containing glucopeptides similar to those from human red blood cells.. Journal of General Microbiology 130:2757–2766
    [Google Scholar]
  22. Patel P.V., Parsons N.J., Andrade J.R.C., Nairn C.A., Tan E.L., Goldner M., Cole J.A., Smith H. 1988; White blood cells including polymorphonuclear phagocytes contain a factor which induces gonococcal resistance to complement- mediated serum killing.. FEMS Microbiology Letters 50:173–176
    [Google Scholar]
  23. Penn C.W., Sbn D., Veale D.R., Parsons N.J., Smith H., Witt K. 1976; Morphological, biological and antigenic properties of Neisseria gonorrhoeae adapted to growth in guinea pig subcutaneous chambers.. Journal of General Microbiology 97:35–43
    [Google Scholar]
  24. Perry M.B., Daoust V., Diena B.B., Ashton F.E., Wallace R. 1975; The lipopolysaccharides of Neisseria gonorrhoeae colony types 1 and 4.. Canadian Journal of Biochemistry 53:623–629
    [Google Scholar]
  25. Rice P.A., Mccormack W.M., Kasper D.L. 1980; Natural serum bactericidal activity against Neisseria gonorrhoeae isolates from disseminated, locally invasive and uncomplicated disease.. Journal of Immunology 124:2105–2109
    [Google Scholar]
  26. Rittenberg S.C., Penn C.W., Parsons N.J., Veale D.R., Smith H. 1977; Phenotypic changes in the resistance of Neisseria gonorrhoeae to killing by normal human serum.. Journal of General Microbiology 103:69–75
    [Google Scholar]
  27. Robbins J.B., Schneerson R., Egan W.G., Vann W., Liu D.T. 1980; Virulence properties of bacterial capsular polysaccharides - unanswered questions.. In The Molecular Basis of Microbial Pathogenicity pp 115–132 Smith H., Skehel J. J., Turner M. J. Edited by Weinheim: Verlag Chemie;
    [Google Scholar]
  28. Schoolnik G.K., Ochs H.D., Buchanan T.M. 1979; Immunoglobulin class responsible for gonococcal bactericidal activity of normal human sera.. Journal of Immunology 122:1771–1779
    [Google Scholar]
  29. Scudder P.R., Chantler E.N. 1981; Glycosyl- transferases of the human cervical epithelium. II. Characteristics of a CMP-JV-acetylneuraminate, galactosyl glycoprotein sialyltransferase.. Biochimka et biophysica acta 660:136–141
    [Google Scholar]
  30. Smith H. 1988; The development of studies on the determinants of bacterial pathogenicity.. Journal of Comparative Pathology 98: in the Press
    [Google Scholar]
  31. Stephens D.S., Shafer W.M. 1987; Evidence that the serum resistance genetic locus sac-3 of Neisseria gonorrhoeae is involved in lipopolysaccharide structure.. Journal of General Mkrobiology 133:2671–2678
    [Google Scholar]
  32. Tan E.L., Patel P.V., Parsons N.J., Martin P.M.V., Smith H. 1986; Lipopolysaccharide alteration is associated with induced resistance of Neisseria gonorrhoeae to killing by human serum.. Journal of General Mkrobiology 132:1407–1413
    [Google Scholar]
  33. Veale D.R., Penn C.W., Smith H. 1981; Factors affecting the induction of phenotypically determined serum resistance of Neisseria gonorrhoeae grown in media containing serum or its diffusible components.. Journal of General Microbiology 122:235–245
    [Google Scholar]
  34. Ward M.E., Watt P.J., Glynn A.A. 1970; Gonococci in urethral exudates possess a virulence factor lost on subculture.. Nature, London 227:382–384
    [Google Scholar]
  35. Weiss J.B., Lote C.J., Bobinski H. 1971; New low molecular weight glycopeptide containing tri- glucosylcysteine in human erythrocyte membrane.. Nature New Biology 234:25–26
    [Google Scholar]
  36. Winstanley F.D., Blackwell C.C., Tan E.L., Patel P.V., Parsons N.J., Martin P.M.V., Smith H. 1984; Alteration of pyocin-sensitivity pattern of Neisseria gonorrhoeae is associated with induced resistance to killing by human serum.. Journal of General Microbiology 130:1303–1306
    [Google Scholar]
  37. Wiseman G.M., Cairo J.D. 1977; Composition of the lipopolysaccharide of Neisseria gonorrhoeae. . Infection and Immunity 16:550–556
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-12-3295
Loading
/content/journal/micro/10.1099/00221287-134-12-3295
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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