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. 1994 Dec;62(12):5327–5334. doi: 10.1128/iai.62.12.5327-5334.1994

Alteration of protective and serologic responses in BALB/c mice vaccinated with chemically modified versus nonmodified proteins of Brucella abortus 19.

G W Pugh Jr 1, L B Tabatabai 1
PMCID: PMC303272  PMID: 7960111

Abstract

A study was conducted to determine whether the covalent chemical modification of Brucella abortus 19 salt-extractable proteins (BCSP) and BCSP derivatives would modulate the immune responses in BALB/c mice. Salt-extractable proteins BCSP 0-70 and BCSP 70-100 were modified with acetoacetic anhydride, and recombinant proteins rBCSP20 (20 kDa), rBCSP31 (31 kDa), and rBCSP45 (45 kDa) were modified with succinic and dodecanoyl anhydrides. Four weeks after mice were vaccinated with the different preparations, principal and control mice were challenge exposed with a virulent culture of B. abortus 2308, and mice were necropsied 2 weeks later. Serum samples were obtained immediately before mice were challenge exposed and at necropsy. Sera were tested for specific immunoglobulin M (IgM) and G (IgG) antibodies by using an enzyme-linked immunosorbent assay. Acylation decreased the immune responses (increased IgG antibodies and reduced spleen CFU and splenomegaly) induced by both BCSP 0-70 and BCSP 70-100. Modification of the recombinant proteins by dodecanoyl and succinic anhydrides had no effect on the protection induced; however, the IgG serologic responses to the homologous and heterologous proteins were altered. Monophosphoryl lipid A markedly enhanced the immunogenicity of BCSP 0-70.

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Selected References

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  1. Allison A. C., Gregoriadis G. Vaccines: recent trends and progress. Immunol Today. 1990 Dec;11(12):427–429. doi: 10.1016/0167-5699(90)90164-5. [DOI] [PubMed] [Google Scholar]
  2. Araya L. N., Elzer P. H., Rowe G. E., Enright F. M., Winter A. J. Temporal development of protective cell-mediated and humoral immunity in BALB/c mice infected with Brucella abortus. J Immunol. 1989 Nov 15;143(10):3330–3337. [PubMed] [Google Scholar]
  3. Beauclair K. D., Khansari D. N. Protection of mice against Brucella abortus by immunization with polyclonal anti-idiotype antibodies. Immunobiology. 1990 Feb;180(2-3):208–220. doi: 10.1016/s0171-2985(11)80329-3. [DOI] [PubMed] [Google Scholar]
  4. Belzer C. A., Tabatabai L. B., Deyoe B. L. Differentiation by western blotting of immune responses of cattle vaccinated with Brucella abortus strain 19 or infected experimentally or naturally with virulent Brucella abortus. Vet Microbiol. 1991 Mar;27(1):79–90. doi: 10.1016/0378-1135(91)90064-m. [DOI] [PubMed] [Google Scholar]
  5. Bosseray N. Immunity to Brucella in mice vaccinated with a fraction (F8) or a killed vaccine (H38) with or without adjuvant. Level and duration of immunity in relation to dose of vaccine, recall injection and age of mice. Br J Exp Pathol. 1978 Aug;59(4):354–365. [PMC free article] [PubMed] [Google Scholar]
  6. Bricker B. J., Tabatabai L. B., Deyoe B. L., Mayfield J. E. Conservation of antigenicity in a 31-kDa Brucella protein. Vet Microbiol. 1988 Dec;18(3-4):313–325. doi: 10.1016/0378-1135(88)90096-x. [DOI] [PubMed] [Google Scholar]
  7. Cheers C. Pathogenesis and cellular immunity in experimental murine brucellosis. Dev Biol Stand. 1984;56:237–246. [PubMed] [Google Scholar]
  8. Confer A. W., Tabatabai L. B., Deyoe B. L., Oltjen S. L., Hall S. M., Oltjen J. W., Morton R. J., Fulnechek D. L., Smith R. E., Smith R. A. Vaccination of cattle with chemically modified and unmodified salt-extractable proteins from Brucella abortus. Vet Microbiol. 1987 Dec;15(4):325–339. doi: 10.1016/0378-1135(87)90020-4. [DOI] [PubMed] [Google Scholar]
  9. Coon J., Hunter R. Properties of conjugated protein immunogens which selectively stimulate delayed-type hypersensitivity. J Immunol. 1975 May;114(5):1518–1522. [PubMed] [Google Scholar]
  10. Dubray G. Protective antigens in brucellosis. Ann Inst Pasteur Microbiol. 1987 Jan-Feb;138(1):84–87. doi: 10.1016/0769-2609(87)90080-9. [DOI] [PubMed] [Google Scholar]
  11. Habeeb A. F. Determination of free amino groups in proteins by trinitrobenzenesulfonic acid. Anal Biochem. 1966 Mar;14(3):328–336. doi: 10.1016/0003-2697(66)90275-2. [DOI] [PubMed] [Google Scholar]
  12. Ho M., Cheers C. Resistance and susceptibility of mice to bacterial infection. IV. Genetic and cellular basis of resistance to chronic infection with Brucella abortus. J Infect Dis. 1982 Sep;146(3):381–387. doi: 10.1093/infdis/146.3.381. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Lynn W. A., Golenbock D. T. Lipopolysaccharide antagonists. Immunol Today. 1992 Jul;13(7):271–276. doi: 10.1016/0167-5699(92)90009-V. [DOI] [PubMed] [Google Scholar]
  15. Marzotto A., Pajetta P., Galzigna L., Scoffone E. Reversible acetoacetylation of amino groups in proteins . Biochim Biophys Acta. 1968 Apr 9;154(3):450–456. doi: 10.1016/0005-2795(68)90004-4. [DOI] [PubMed] [Google Scholar]
  16. Montaraz J. A., Winter A. J. Comparison of living and nonliving vaccines for Brucella abortus in BALB/c mice. Infect Immun. 1986 Aug;53(2):245–251. doi: 10.1128/iai.53.2.245-251.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Moreno E., Kurtz R. S., Berman D. T. Induction of immune and adjuvant immunoglobulin G responses in mice by Brucella lipopolysaccharide. Infect Immun. 1984 Oct;46(1):74–80. doi: 10.1128/iai.46.1.74-80.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Naor D., Galili N. Immune response to chemically modified antigens. Prog Allergy. 1977;22:107–146. [PubMed] [Google Scholar]
  19. Parish C. R. Immune response to chemically modified flagellin. II. Evidence for a fundamental relationship between humoral and cell-mediated immunity. J Exp Med. 1971 Jul 1;134(1):21–47. doi: 10.1084/jem.134.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Parish C. R. The relationship between humoral and cell-mediated immunity. Transplant Rev. 1972;13:35–66. doi: 10.1111/j.1600-065x.1972.tb00059.x. [DOI] [PubMed] [Google Scholar]
  21. Phillips M., Deyoe B. L., Canning P. C. Protection of mice against Brucella abortus infection by inoculation with monoclonal antibodies recognizing Brucella O-antigen. Am J Vet Res. 1989 Dec;50(12):2158–2161. [PubMed] [Google Scholar]
  22. Phillips M., Pugh G. W., Jr, Deyoe B. L. Chemical and protective properties of Brucella lipopolysaccharide obtained by butanol extraction. Am J Vet Res. 1989 Mar;50(3):311–317. [PubMed] [Google Scholar]
  23. Plommet M. Brucellosis and immunity: humoral and cellular components in mice. Ann Inst Pasteur Microbiol. 1987 Jan-Feb;138(1):105–110. doi: 10.1016/0769-2609(87)90086-x. [DOI] [PubMed] [Google Scholar]
  24. Pugh G. W., Jr, Tabatabai L. B., Bricker B. J., Mayfield J. E., Phillips M., McDonald T. J., Zehr E. S. Identification of a virulence factor for Brucella abortus infection in BALB/c mice. Am J Vet Res. 1989 Jun;50(6):887–892. [PubMed] [Google Scholar]
  25. Pugh G. W., Jr, Tabatabai L. B., Bricker B. J., Mayfield J. E., Phillips M., Zehr E. S., Belzer C. A. Immunogenicity of Brucella-extracted and recombinant protein vaccines in CD-1 and BALB/c mice. Am J Vet Res. 1990 Sep;51(9):1413–1420. [PubMed] [Google Scholar]
  26. Pugh G. W., Jr, Tabatabai L. B., Phillips M., McDonald T. J. Establishment of dose-response relationships in BALB/c mice, using Brucella cell surface protein and lipopolysaccharide. Am J Vet Res. 1991 Feb;52(2):261–268. [PubMed] [Google Scholar]
  27. Rubin B., Aasted B. Characterization of new antigenic determinants introduced into homologous serum albumin by dinitrophenylation and sulphanylation. Immunology. 1973 Sep;25(3):399–408. [PMC free article] [PubMed] [Google Scholar]
  28. Schirrmacher V., Wigzell H. Immune responses against native and chemically modified albumins in mice. II. Effect of electric charge and conformation on the humoral antibody response and on helper T cell responses. J Immunol. 1974 Nov;113(5):1635–1643. [PubMed] [Google Scholar]
  29. Splitter G. A., Everlith K. M. Brucella abortus regulates bovine macrophage-T-cell interaction by major histocompatibility complex class II and interleukin-1 expression. Infect Immun. 1989 Apr;57(4):1151–1157. doi: 10.1128/iai.57.4.1151-1157.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tabatabai L. B., Deyoe B. L. Biochemical and biological properties of soluble protein preparations from Brucella abortus. Dev Biol Stand. 1984;56:199–211. [PubMed] [Google Scholar]
  31. Tabatabai L. B., Deyoe B. L. Characterization of salt-extractable protein antigens from Brucella abortus by crossed immunoelectrophoresis and isoelectricfocusing. Vet Microbiol. 1984 Oct;9(6):549–560. doi: 10.1016/0378-1135(84)90017-8. [DOI] [PubMed] [Google Scholar]
  32. Tabatabai L. B., Deyoe B. L., Patterson J. M. Immunogenicity of Brucella abortus salt-extractable proteins. Vet Microbiol. 1989 May;20(1):49–58. doi: 10.1016/0378-1135(89)90006-0. [DOI] [PubMed] [Google Scholar]
  33. Tabatabai L. B., Deyoe B. L. Specific enzyme-linked immunosorbent assay for detection of bovine antibody to Brucella abortus. J Clin Microbiol. 1984 Aug;20(2):209–213. doi: 10.1128/jcm.20.2.209-213.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tabatabai L. B., Pugh G. W., Jr, Stevens M. G., Phillips M., McDonald T. J. Monophosphoryl lipid A-induced immune enhancement of Brucella abortus salt-extractable protein and lipopolysaccharide vaccines in BALB/c mice. Am J Vet Res. 1992 Oct;53(10):1900–1907. [PubMed] [Google Scholar]
  35. 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 Mar;18(2):149–163. doi: 10.1016/0165-2427(88)90057-8. [DOI] [PubMed] [Google Scholar]
  36. Young E. J. Brucellosis: a model zoonosis in developing countries. APMIS Suppl. 1988;3:17–20. [PubMed] [Google Scholar]

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