1887

Abstract

Summary: Twenty-two monoclonal antibodies (mAbs) specific for smooth lipopolysaccharide (S-LPS) were generated by fusion of spleen cells from mice infected with the rough strain B115 with the NSO myeloma. According to reactivity in enzyme-linked immunosorbent assay (ELISA) with O-polysaccharide (O-PS) and absence of reactivity with rough lipopolysaccharide (R-LPS), it was postulated that these mAbs recognized epitopes present on the O-PS. Most of the mAbs reacted equally well in ELISA and immunoblotting with S-LPS types of Brucella A and M dominant strains and were designated as specific for common (C) epitopes. Three mAbs were highly specific for M dominant S-LPS. All these mAbs, in contrast to a mAb specific for the A epitope, showed little or no cross-reactivity with O:9 S-LPS. S-LPS of B115 was extracted and analysed by immunoblotting and ELISA with mAbs specific for A, M and C epitopes. Reactivity of the mAbs with this S-LPS was compared to reactivity with S-LPS of A and M dominant smooth strains. The results suggest that S-LPS of B115 bears mainly C epitopes and a few M epitopes. The very weak reactivity of this S-LPS with the mAb specific for the A epitope and the fact that the mAbs specific for C and M epitopes showed little or no cross-reactivity with O:9 S-LPS suggest that O-PS from this rough strain could be used to distinguish O:9 infection from infection. The potential value of the rough strain B115 as a vaccine strain is also discussed.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-7-1551
1993-07-01
2021-05-06
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/7/mic-139-7-1551.html?itemId=/content/journal/micro/10.1099/00221287-139-7-1551&mimeType=html&fmt=ahah

References

  1. Alton G.G. 1960; The occurrence of dissociated strains of Brucella melitensis in the milk of goats in Malta.. Journal of Comparative Pathology 70:10–17
    [Google Scholar]
  2. Alton G.G., Jones L.M., Angus R.D., Verger J.M. 1988 Techniques for the Brucellosis Laboratory. Paris:: Institut National de la Recherche Agronomique.;
    [Google Scholar]
  3. Bundle D.R., Cherwonogrodzky J.W., Gidney M.A.J., Meikle P.J., Perry M.B., Peters T. 1989; Definition of Brucella A and M epitopes by monoclonal typing reagents and synthetic oligosaccharides.. Infection and Immunity 57:2829–2836
    [Google Scholar]
  4. Cloeckaert A., De Wergifosse P., Dubray G., Limet J.N. 1990; Identification of seven surface-exposed Brucella outer membrane proteins by use of monoclonal antibodies: immunogold labeling for electron microscopy and enzyme-linked immunosorbent assay.. Infection and Immunity 58:3980–3987
    [Google Scholar]
  5. Cloeckaert A., Jacques I., De Wergifosse P., Dubray G., Limet J.N. 1992a; Protection against Brucella melitensis or Brucella abortus in mice with immunoglobulin G (IgG), IgA and IgM monoclonal antibodies specific for a common epitope shared by the Brucella A and M smooth lipopolysaccharides.. Infection and Immunity 60:312–315
    [Google Scholar]
  6. Cloeckaert A., Zygmunt M.S., Nicolle J.-C., Dubray G., Limet J.N. 1992b; O-chain expression in the rough Brucella melitensis strain B115: induction of O-polysaccharide specific monoclonal antibodies and intracellular localization demonstrated byimmunoelectron microscopy.. Journal of General Microbiology 138:1211–1219
    [Google Scholar]
  7. Douglas J.T., Palmer D.A. 1988; Use of monoclonal antibodies to identify the distribution of A and M epitopes on smooth Brucellaspecies.. Journal of Clinical Microbiology 26:1353–1356
    [Google Scholar]
  8. Dubray G. 1981 Etude ultrastructurale et biochimique des enveloppes des bactéries du genre Brucella. PhD thesis Université de Paris-Sud.:
    [Google Scholar]
  9. Galanos C., Luderitz O., Westphal O. 1969; A new method for the extraction of R-lipopolysaccharides.. European Journal of Biochemistry 9:245–249
    [Google Scholar]
  10. Garin-Bastuji B., Bowden R.A., Dubray G., Limet J.N. 1990; Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting analysis of smooth-lipopolysaccharide hetero-geneity among Brucella biovars related to A and M specificities.. Journal of Clinical Microbiology 28:2169–2174
    [Google Scholar]
  11. Harmon B.G., Adams L.G., Frey M. 1988; Survival of rough and smooth strains of Brucella abortus in bovine mammary gland macrophages.. American Journal of Veterinary Research 49:1092–1097
    [Google Scholar]
  12. Jacques I., Olivier-Bernardin V., Dubray G. 1991; Induction of antibody and protective responses in mice by Brucella O-polysaccharide-BSA conjugate.. Vaccine 9:896–900
    [Google Scholar]
  13. Jones L.M., Diaz R., Taylor A.G. 1973; Characterization of allergens prepared from smooth and rough strains of Brucella melitensis.. British Journal of Experimental Pathology 54:492–508
    [Google Scholar]
  14. Kreutzer D.L., Robertson D.C. 1979; Surface macromolecules and virulence in intracellularparasitism: comparison of cell envelope components of smooth and rough strains of Brucella abortus.. Infection and Immunity 23:819–837
    [Google Scholar]
  15. Leong D., Diaz R., Milner K., Rudbach J., Wilson J.B. 1970; Some structural and biological properties of Brucella endotoxin.. Infection and Immunity 1:174–182
    [Google Scholar]
  16. Limet J.N., Berbinschi A., Cloeckaert A., Cambiaso C.L., Masson P.L. 1988; Longitudinal study of brucellosis in mice by immunoassay of lipopolysaccharide-related antigens in blood and urine.. Journal of Medical Microbiology 26:37–45
    [Google Scholar]
  17. Limet J.N., Bosseray N., Garin-Bastuji B., Dubray G., Plommet M. 1989; Humoral immunity in mice mediated by monoclonal antibodies against the A and M antigens of Brucella.. Journal of Medical Microbiology 30:37–43
    [Google Scholar]
  18. Meikle P.J., Perry M.B., Cherwonogrodzky J.W., Bundle D.R. 1989; Fine structure of A and M antigens from Brucellabiovars.. Infection and Immunity 57:2820–2828
    [Google Scholar]
  19. Palmer D.A., Douglas J.T. 1989; Analysis of Brucellalipopolysaccharide with specific and cross-reacting monoclonal antibodies.. Journal of Clinical Microbiology 27:2331–2337
    [Google Scholar]
  20. Roop R.M. II Preston-Moore D., Bagchi T., Schurig G.G. 1987; Rapid identification of smooth Brucella species with a monoclonal antibody.. Journal of Clinical Microbiology 25:2090–2093
    [Google Scholar]
  21. Schurig G.G., Roop R.M. II Bagchi T., Boyle S., Buhrman D., Sriranganathan N. 1991; Biological properties of RB51; a stable rough strain of Brucella abortus.. Veterinary Microbiology 28:171–188
    [Google Scholar]
  22. Tobias L., Schurig G.G., Cordes D.O. 1992; Comparative behaviour of Brucella abortus strains 19 and RB51 in the pregnant mouse.. Research in Veterinary Science 53:179–183
    [Google Scholar]
  23. Vizcaino N., Fernandez-Lago L. 1992; A rapid and sensitive method for the identification of Brucella species with a monoclonal antibody.. Research in Microbiology 143:513–518
    [Google Scholar]
  24. Wilson G.S., Miles A.A. 1932; The serological differentiation of smooth strains of the Brucella group.. British Journal of Experimental Pathology 13:1–13
    [Google Scholar]
  25. Zygmunt M.S., Dubray G., Bundle D.R., Perry M.B. 1988; Purified native haptens of Brucella abortus B19 and B. melitensis16M reveal the lipopolysaccharide origin of the antigens.. Annales de l̓Institut Pasteur/Microbiologie 139:421–433
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-7-1551
Loading
/content/journal/micro/10.1099/00221287-139-7-1551
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