1887

Journal of Medical Microbiology logo

Volume 47, Issue 1

Evaluation Of Immunogenicity and Protective Activity in Balb/C Mice of the 25-kDa Major Outer-Membrane Protein Of (Omp25) Expressed in Free

Abstract

The antibody response specific to the 25-kDa major outer-membrane protein (Omp25) of expressed in was assessed in BALB/c mice. Groups of mice were immunised and boosted either with sonicated carrying plasmid pAC2533 – (pAC2533) – expressing the gene coding for Omp25 (25 gene) of , or with carrying plasmid pUC19 – (pUC19). One control group received saline. The evolution of antibody responses was investigated by indirect ELISA with whole rough (R) H38 cells as antigen. Serum antibody titres of mice immunised with (pAC2533) were appreciably higher than those of mice immunised with (pUC19). The specificity to Omp25 of murine antibodies induced by (pAC2533) was demonstrated by SDS-PAGE and immunoblotting of five strains. Binding of antibody in (pAC2533) immune sera to the surface of strains differing in their smooth lipopolysaccharide (S-LPS) expression was also studied by whole-cell ELISA and by flow cytometry. Antibody reactivity to R and smooth-rough (S-R) was much stronger than that to smooth (S) strains, indicating a much better accessibility of Omp25 to antibody on strains lacking or expressing less O-polysaccharide on their surface. The antibodies to Omp25 were predominantly of IgG2a isotype. The capacity of (pAC2533) to induce protective immune responses against four challenge strains of was further evaluated in mice. Significant reductions in splenic infections, in comparison with mice immunised with (pUC19) and unimmunised (saline injection) mice, were observed in R B115, S-R EP and S H38 infected mice. Protection against S 16M was not significant. The data from the present study, together with previous results, suggest that humoral immunity against probably conformational, well-exposed epitopes of the Omp25 could contribute to protective mechanisms against infection in mice.

  • Received:
  • Accepted:
  • Published Online:
© 1998 The Pathological Society of Great Britain and Ireland
Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-47-1-39
1998-01-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/jmm/47/1/medmicro-47-1-39.html?itemId=/content/journal/jmm/10.1099/00222615-47-1-39&mimeType=html&fmt=ahah

References

  1. Adams L. G. Development of live Brucella vaccines. In Adams L. G. (ed) Advances in brucellosis research College Station, Texas A & M University Press. TX; 1990250–276
     [Google Scholar]
  2. Plommet M. Killed vaccines in cattle: current situation and perspectives. In Adams L. G. (ed) Advances in brucellosis research College Station, Texas A & M University Press. TX; 1990215–227
     [Google Scholar]
  3. Dubray G., Bezard G. Isolation of three Brucella abortus cell-wall antigens protective in murine experimental brucellosis. Ann Rech Vet 1980; 11:367–373
     [Google Scholar]
  4. Dubray G., Charriaut C. Evidence of three major polypeptide species and two major polysaccharide species in the Brucella outer membrane. Ann Rech Vet 1983; 14:311–318
     [Google Scholar]
  5. Dubray G. Protective antigens in brucellosis. Ann Inst Pasteur Microbiol 1987; 138:84–87
     [Google Scholar]
  6. Jimenez de Bagüés M. P., Elzer P. H., Blasco J. M., Marin C. M., Gamazo C., Winter A. J. Protective immunity to Brucella ovis in BALB/c mice following recovery from primary infection or immunization with subcellular vaccines. Infect Immun 1994; 62:632–638
     [Google Scholar]
  7. Winter A. J., Rowe G. E. Comparative immune responses to native cell envelope antigens and the hot sodium dodecyl sulfate insoluble fraction (PG) of Brucella abortus in cattle and mice. Vet Immunol Immunopathol 1988; 18:149–163
     [Google Scholar]
  8. Robinson A., Melling J. Envelope structure and the development of new vaccines. J Appl Bacteriol 1993; 74: Suppl 43S–51S
     [Google Scholar]
  9. Cloeckaert A., Jacques I., Limet J. N., Dubray G. Immunogenic properties of Brucella melitensis cell-wall fractions in BALB/c mice. J Med Microbiol 1995; 42:200–208
     [Google Scholar]
  10. Sowa B. A., Kelly K. A., Ficht T. A., Frey M., Adams L. G. SDS-soluble and peptidoglycan-bound proteins in the outer mem-brane-peptidoglycan complex of Brucella abortus. Vet Microbiol 1991; 27:351–369
     [Google Scholar]
  11. Gamazo C., Winter A. J., MoriyÓn I., Riezu-Boj J. I., Blasco J. M., Diaz R. Comparative analyses of proteins extracted by hot saline or released spontaneously into outer membrane blebs from field strains of Brucella ovis and Brucella melitensis. Infect Immun 1989; 57:1419–1426
     [Google Scholar]
  12. Cloeckaert A., Zygmunt M. S., de Wergifosse P., Dubray G., Limet J. N. Demonstration of peptidoglycan-associated Brucella outer-membrane proteins by use of monoclonal antibodies. J Gen Microbiol 1992; 138:1543–1550
     [Google Scholar]
  13. Verstreate D. R., Creasy M. T., Caveney N. T., Baldwin C. L., Blab M. W., Winter A. J. Outer membrane proteins of Brucella abortus: isolation and characterization. Infect Immun 1982; 35:979–989
     [Google Scholar]
  14. Cherwonogrodzky J. W., Dubray G., Moreno E., Mayer H. Antigens of Brucella. In Nielsen K., Duncan J. R. (eds) Animal brucellosis Boca Raton: CRC Press; 199019–64
     [Google Scholar]
  15. MoriyÓn I., Gamazo C., Diaz R. Properties of the outer membrane of Brucella. Ann Inst Pasteur Microbiol 1987; 138:89–91
     [Google Scholar]
  16. Cloeckaert A., Jacques I., de Wergifosse P., Dubray G., Limet J. N. 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. Infect Immun 1992; 60:312–315
     [Google Scholar]
  17. Limet J., Plommet A. M., Dubray G., Plommet M. Immunity conferred upon mice by anti-LPS monoclonal antibodies in murine brucellosis. Ann Inst Pasteur Immunol 1987; 138:417–424
     [Google Scholar]
  18. Limet J. N., Bosseray N., Garin-Bastuji B., Dubray G., Plommet M. Humoral immunity in mice mediated by monoclonal antibodies directed against the A and M antigens of Brucella. J Med Microbiol 1989; 30:37–43
     [Google Scholar]
  19. Montaraz J. A., Winter A. J., Hunter D. M., Sowa B. A., Wu A. M., Adams L. G. Protection against Brucella abortus in mice with O-polysaccharide-specific monoclonal antibodies. Infect Immun 1986; 51:961–963
     [Google Scholar]
  20. Phillips M., Deyoe B., Canning P. C. Protection of mice against Brucella abortus infection by inoculation with monoclonal antibodies recognizing Brucella O-antigen. Am J Vet Res 1989; 50:2158–2161
     [Google Scholar]
  21. Cloeckaert A., Jacques I., Bosseray N. Protection conferred on mice by monoclonal antibodies directed against outer-membrane-protein antigens of Brucellal. J Med Microbiol 1991; 34:175–180
     [Google Scholar]
  22. Cloeckaer A., Jacques I., Bowden R. A., Dubray G., Limet J. N. Monoclonal antibodies to Brucella rough lipopolysaccharide: characterization and evaluation of their protective effect against B. abortus. Res Microbiol 1993; 144:475–484
     [Google Scholar]
  23. Jacques I., Cloeckaert A., Limet J. N., Dubray G. Protection conferred on mice by combinations of monoclonal antibodies directed against outer-membrane proteins or smooth lipopolysaccharide of Brucella. J Med Microbiol 1992; 37:100–103
     [Google Scholar]
  24. Bowden R. A., Cloeckaert A., Zygmunt M. S., Bernard S., Dubray G. Surface exposure of outer membrane protein and lipopolysaccharide epitopes in Brucella species studied by enzyme-linked immunosorbent assay and flow cytometry. Infect Immun 1995; 63:3945–3952
     [Google Scholar]
  25. Cloeckaert A., de Wergifosse P., Dubray G., Limet J. N. Identification of seven surface-exposed Brucella outer membrane proteins by use of monoclonal antibodies: immunogold labeling for electron microscopy and enzyme-linked immunosorbent assay. Infect Immun 1990; 58:3980–3987
     [Google Scholar]
  26. de Wergifosse P., Lintermans P., Limet J. N., Cloeckaert A. Cloning and nucleotide sequence of the gene cloding for the major 25-kilodalton outer membrane protein of Brucella abortus. J Bacterial 1995; 177:1911–1914
     [Google Scholar]
  27. Cloeckaert A., Verger J. M., Grayon M., Grepinet O. Restriction site polymorphism of the genes encoding the major 25 kDa and 36 kDa outer-membrane proteins of Brucella. Microbiology 1995; 141:2111–2121
     [Google Scholar]
  28. Cloeckaert A., Verger J. M., Grayon M., Zygmunt M. S., Grepinet O. Nucleotide sequence and expression of the gene encoding the major 25-kilodalton outer membrane protein of Brucella ovis: evidence for antigenic shift, compared with other Brucella species, due to a deletion in the gene. Infect Immun 1996; 64:2047–2055
     [Google Scholar]
  29. Alton G. G., Jones L. M., Angus R. D., Verger J. M. Techniques for the brucellosis laboratory. Paris: Institut National de la Recherche Agronomique; 1988
     [Google Scholar]
  30. Bowden R. A., Verger J. M., Grayon M., Limet J. N., Dubray G. Simultaneous expression of smooth and rough phase properties related to lipopolysaccharide in a strain of Brucella melitensis. J Med Microbiol 1993; 39:363–370
     [Google Scholar]
  31. Cloeckaert A., Zygmunt M. S., Nicolle J. C., Dubray G., Limet J. N. O-chain expression in the rough Brucella melitensis strain B115: induction of O-polysaccharide-specific monoclonal antibodies and intracellular localization demonstrated by immuno-electron microscopy. J Gen Microbiol 1992; 138:1211–1219
     [Google Scholar]
  32. Mossman T. R., Fong T. A. T. Specific assays for cytokine production by T cells. J Immunol Methods 1989; 116:151–158
     [Google Scholar]
  33. Tibor A., Weynants V., Denoel P. Molecular cloning, nucleotide sequence, and occurrence of a 16.5 kilodalton outer membrane protein of Brucella abortus with similarity to PAL lipoproteins. Infect Immun 1994; 62:3633–3639
     [Google Scholar]
  34. Berzofsky J. A., Berkower I. J. Immunogenicity and antigen structure. In Paul W. E. (ed) Fundamental immunology 2nd edn New York: Raven Press; 1989169–208
     [Google Scholar]
  35. Cloeckaert A., Zygmunt M. S., Bezard G., Dubray G. Purification and antigenic analysis of the major 25-kilodalton outer membrane protein of Brucella abortus. Res Microbiol 1996; 147:225–235
     [Google Scholar]
  36. Cardenas L., Clements J. D. Stability, immunogenicity and expression of foreign antigens in bacterial vaccine vectors. Vaccine 1993; 11:126–135
     [Google Scholar]
  37. Leclerc C., Charbit A., Molla A., Hofnung M. Antibody response to a foreign epitope expressed at the surface of recombinant bacteria: importance of the route of immunization. Vaccine 1989; 7:242–248
     [Google Scholar]
  38. Wetzler L. M., Blake M. S., Barry K., Gotschlich E. C. Gonococcal porin vaccine evaluation: comparison of por proteosomes, liposomes, and blebs isolated from rmp deletion mutants. J Infect Dis 1992; 166:551–555
     [Google Scholar]
  39. Muthukkumar S., Muthukkaruppan V. R. Mechanism of protective immunity induced by porin-lipopolysaccharide against murine salmonellosis. Infect Immun 1993; 61:3017–3025
     [Google Scholar]
  40. Nurminen M., Butcher S., Idanpaan-Heikkila I. The class 1 outer membrane protein of Neisseria meningitidis produced in Bacillus subtilis can give rise to protective immunity. Mol Microbiol 1992; 6:2499–2506
     [Google Scholar]
  41. Moriyon I., Montanes M. A. In vitro interactions between lipopolysaccharides and heterologous outer membrane porin proteins. Curr Microbiol 1985; 12:229–234
     [Google Scholar]
  42. Winter A. J., Schurig G. G., Boyle S. M. Protection of BALB/c mice against homologous and heterologous species of Brucella by rough strain vaccines derived from Brucella melitensis and Brucella suis biovar 4. Am J Vet Res 1996; 57:677–683
     [Google Scholar]
  43. Jones S. M., Winter A. J. Survival of virulent and attenuated strains of Brucella abortus in normal and gamma interferon-activated murine peritoneal macrophages. Infect Immun 1992; 60:3011–3014
     [Google Scholar]
  44. Zhan Y., Cheers C. Endogenous gamma interferon mediates resistance to Brucella abortus infection. Infect Immun 1993; 61:4899–4901
     [Google Scholar]
  45. ten Hagen T. L. M., Sulzer A. J., Kidd M. R., Lai A. A., Hunter R. L. Role of adjuvants in the modulation of antibody isotype, specificity, and induction of protection by whole blood-stage Plasmodium yoelii vaccines. J Immunol 1993; 151:7077–7085
     [Google Scholar]
  46. Finkelman F. D., Katona I. M., Mosmann T. R., Coffman R. L. IFN-γ regulates the isotype of Ig secreted during in vivo humoral immune responses. J Immunol 1988; 140:1022–1027
     [Google Scholar]
  47. Potter M. Immunoglobulins and immunoglobulin genes. In Foster H. L., Small J. D., Fox J. (eds) Mouse in biomedical research vol 2 New York: Academic Press; 1984347–380
     [Google Scholar]
  48. Bowden R. A., Cloeckaert A., Zygmunt M. S., Dubray G. Outer-membrane protein-and rough lipopolysaccharide-specific monoclonal antibodies protect mice against Brucella ovis. J Med Microbiol 1995; 43:344–347
     [Google Scholar]
  49. Smith R., Adams L. G., Ficht T. A., Sowa B. A., Wu A. Immunogenicity of subcellular fractions of Brucella abortus: measurement by in vitro lymphocyte proliferative responses. Vet Immunol Immunopathol 1990; 25:83–97
     [Google Scholar]
  50. Stabel T. J., Mayfield J. E., Tabatabai L. B., Wannemuehler M. J. Oral immunization of mice with an attenuated Salmonella typhi-murium containing a recombinant plasmid which codes for production of a 31-kilodalton protein of Brucella abortus. Infect Immun 1990; 58:2048–2055
     [Google Scholar]
  51. Stabel T. J., Mayfield J. E., Morfitt D. C., Wannemuehler M. J. Oral immunization of mice and swine with an attenuated Salmonella choleraesuis [Acya-12A (crp-cdt)19] mutant containing a recombinant plasmid. Infect Immun 1993; 61:610–618
     [Google Scholar]
  52. Tabatabai L. B., Pugh G. W. Modulation of immune responses in Balb/c mice vaccinated with Brucella abortus Cu-Zn superoxide dismutase synthetic peptide vaccine. Vaccine 1994; 12:919–924
     [Google Scholar]
  53. Oliveira S. C., Splitter G. A. Immunization of mice with recombinant L7/L12 ribosomal protein confers protection against Brucella abortus infection. Vaccine 1996; 14:959–962
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-47-1-39
Loading
Evaluation Of Immunogenicity and Protective Activity in Balb/C Mice of the 25-kDa Major Outer-Membrane Protein Of Brucella Melitensis (Omp25) Expressed in Escherichia Coli
J. Med. Microbiol. 47, 39 (1998); https://doi.org/10.1099/00222615-47-1-39
/content/journal/jmm/10.1099/00222615-47-1-39
/content/journal/jmm/10.1099/00222615-47-1-39
Loading

Data & Media loading...

Most cited 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