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. 1994 Nov;62(11):4990–4996. doi: 10.1128/iai.62.11.4990-4996.1994

Vaccination with Brucella abortus rough mutant RB51 protects BALB/c mice against virulent strains of Brucella abortus, Brucella melitensis, and Brucella ovis.

M P Jiménez de Bagüés 1, P H Elzer 1, S M Jones 1, J M Blasco 1, F M Enright 1, G G Schurig 1, A J Winter 1
PMCID: PMC303217  PMID: 7927779

Abstract

Vaccination of BALB/c mice with live Brucella abortus RB51, a stable rough mutant, produced protection against challenge with virulent strains of Brucella abortus, Brucella melitensis, and Brucella ovis. Passive-transfer experiments indicated that vaccinated mice were protected against B. abortus 2308 through cell-mediated immunity, against B. ovis PA through humoral immunity, and against B. melitensis 16M through both forms of immunity. Live bacteria were required for the induction of protective cell-mediated immunity; vaccination with whole killed cells of strain RB51 failed to protect mice against B. abortus 2308 despite development of good delayed-type hypersensitivity reactions. Protective antibodies against the heterologous species were generated in vaccinated mice primarily through anamnestic responses following challenge infections. Growth of the antigenically unrelated bacterium Listeria monocytogenes in the spleens of vaccinated mice indicated that nonspecific killing by residual activated macrophages contributed minimally to protection. These results encourage the continued investigation of strain RB51 as an alternative vaccine against heterologous Brucella species. However, its usefulness against B. ovis would be limited if, as suggested here, epitopes critical for protective cell-mediated immunity are not shared between B. abortus and B. ovis.

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Selected References

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  1. Afzal M., Tengerdy R. P., Ellis R. P., Kimberling C. V., Morris C. J. Protection of rams against epididymitis by a Brucella ovis-vitamin E adjuvant vaccine. Vet Immunol Immunopathol. 1984 Oct;7(3-4):293–304. doi: 10.1016/0165-2427(84)90087-4. [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. Araya L. N., Winter A. J. Comparative protection of mice against virulent and attenuated strains of Brucella abortus by passive transfer of immune T cells or serum. Infect Immun. 1990 Jan;58(1):254–256. doi: 10.1128/iai.58.1.254-256.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blasco J. M., Díaz R. Brucella melitensis Rev-1 vaccine as a cause of human brucellosis. Lancet. 1993 Sep 25;342(8874):805–805. doi: 10.1016/0140-6736(93)91571-3. [DOI] [PubMed] [Google Scholar]
  5. Caroff M., Bundle D. R., Perry M. B. Structure of the O-chain of the phenol-phase soluble cellular lipopolysaccharide of Yersinia enterocolitica serotype O:9. Eur J Biochem. 1984 Feb 15;139(1):195–200. doi: 10.1111/j.1432-1033.1984.tb07994.x. [DOI] [PubMed] [Google Scholar]
  6. Cheers C., Pagram F. Macrophage activation during experimental murine brucellosis: a basis for chronic infection. Infect Immun. 1979 Feb;23(2):197–205. doi: 10.1128/iai.23.2.197-205.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cheville N. F., Jensen A. E., Halling S. M., Tatum F. M., Morfitt D. C., Hennager S. G., Frerichs W. M., Schurig G. Bacterial survival, lymph node changes, and immunologic responses of cattle vaccinated with standard and mutant strains of Brucella abortus. Am J Vet Res. 1992 Oct;53(10):1881–1888. [PubMed] [Google Scholar]
  8. Cheville N. F., Stevens M. G., Jensen A. E., Tatum F. M., Halling S. M. Immune responses and protection against infection and abortion in cattle experimentally vaccinated with mutant strains of Brucella abortus. Am J Vet Res. 1993 Oct;54(10):1591–1597. [PubMed] [Google Scholar]
  9. Claxton P. D. Brucella ovis vaccination of rams. A comparison of two commercial vaccines and two methods of vaccination. Aust Vet J. 1968 Feb;44(2):48–54. doi: 10.1111/j.1751-0813.1968.tb04953.x. [DOI] [PubMed] [Google Scholar]
  10. Cloeckaert A., Jacques I., Bosseray N., Limet J. N., Bowden R., Dubray G., Plommet M. Protection conferred on mice by monoclonal antibodies directed against outer-membrane-protein antigens of Brucella. J Med Microbiol. 1991 Mar;34(3):175–180. doi: 10.1099/00222615-34-3-175. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Corner L. A., Alton G. G. Persistence of Brucella abortus strain 19 infection in adult cattle vaccinated with reduced doses. Res Vet Sci. 1981 Nov;31(3):342–344. [PubMed] [Google Scholar]
  13. Douglas J. T., Palmer D. A. Use of monoclonal antibodies to identify the distribution of A and M epitopes on smooth Brucella species. J Clin Microbiol. 1988 Jul;26(7):1353–1356. doi: 10.1128/jcm.26.7.1353-1356.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Elzer P. H., Rowe G. E., Enright F. M., Winter A. J. Effects of gamma radiation and azathioprine on Brucella abortus infection in BALB/c mice. Am J Vet Res. 1991 Jun;52(6):838–844. [PubMed] [Google Scholar]
  15. Enright F. M., Araya L. N., Elzer P. H., Rowe G. E., Winter A. J. Comparative histopathology in BALB/c mice infected with virulent and attenuated strains of Brucella abortus. Vet Immunol Immunopathol. 1990 Oct;26(2):171–182. doi: 10.1016/0165-2427(90)90065-z. [DOI] [PubMed] [Google Scholar]
  16. Gamazo C., Winter A. J., Moriyón I., Riezu-Boj J. I., Blasco J. M., Díaz 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 May;57(5):1419–1426. doi: 10.1128/iai.57.5.1419-1426.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Garcia-Carrillo C. Protection of rams against Brucella ovis infection by Brucella melitensis Rev. 1 vaccine. Zentralbl Veterinarmed B. 1981;28(6):425–431. doi: 10.1111/j.1439-0450.1981.tb01762.x. [DOI] [PubMed] [Google Scholar]
  18. Hurvell B., Lindberg A. A. Serological cross-reactions between different Brucella species and Yersinia enterocolitica. Immunochemical studies on phenol-water extracted lipopolysaccharides from Brucella abortus and Yersinia enterocolitica type IX. Acta Pathol Microbiol Scand B Microbiol Immunol. 1973 Feb;81(1):113–119. [PubMed] [Google Scholar]
  19. Jiménez de Bagués M. P., Marin C. M., Barberán M., Blasco J. M. Responses of ewes to B. melitensis Rev1 vaccine administered by subcutaneous or conjunctival routes at different stages of pregnancy. Ann Rech Vet. 1989;20(2):205–213. [PubMed] [Google Scholar]
  20. Jiménez de Bagüs M. P., Elzer P. H., Blasco J. M., Marín 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 Feb;62(2):632–638. doi: 10.1128/iai.62.2.632-638.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jiménez de Bagüs M. P., Marín C. M., Blasco J. M., Moriyón I., Gamazo C. An ELISA with Brucella lipopolysaccharide antigen for the diagnosis of B. melitensis infection in sheep and for the evaluation of serological responses following subcutaneous or conjunctival B. melitensis strain Rev 1 vaccination. Vet Microbiol. 1992 Feb;30(2-3):233–241. doi: 10.1016/0378-1135(92)90117-c. [DOI] [PubMed] [Google Scholar]
  22. 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 Jul;60(7):3011–3014. doi: 10.1128/iai.60.7.3011-3014.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kaneene J. M., Anderson R. K., Johnson D. W., Muscoplat C. C. Brucella antigen preparations for in vitro lymphocyte immunostimulation assays in bovine brucellosis. Infect Immun. 1978 Nov;22(2):486–491. doi: 10.1128/iai.22.2.486-491.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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 May-Jun;138(3):417–424. doi: 10.1016/s0769-2625(87)80052-1. [DOI] [PubMed] [Google Scholar]
  25. McGowan B., Harrold D. R. Epididymitis in rams: studies on vaccine efficacy. Cornell Vet. 1979 Jan;69(1):73–76. [PubMed] [Google Scholar]
  26. 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]
  27. 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 Mar;51(3):961–963. doi: 10.1128/iai.51.3.961-963.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moreno E., Pitt M. W., Jones L. M., Schurig G. G., Berman D. T. Purification and characterization of smooth and rough lipopolysaccharides from Brucella abortus. J Bacteriol. 1979 May;138(2):361–369. doi: 10.1128/jb.138.2.361-369.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ray W. C. An assessment of investigations conducted in the USA on Brucella abortus strain 45/20 bacterins. Dev Biol Stand. 1976;31:335–342. [PubMed] [Google Scholar]
  30. Riezu-Boj J. I., Moriyón I., Blasco J. M., Gamazo C., Díaz R. Antibody response to Brucella ovis outer membrane proteins in ovine brucellosis. Infect Immun. 1990 Feb;58(2):489–494. doi: 10.1128/iai.58.2.489-494.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Riezu-Boj J. I., Moriyón I., Blasco J. M., Marín C. M., Diaz R. Comparison of lipopolysaccharide and outer membrane protein-lipopolysaccharide extracts in an enzyme-linked immunosorbent assay for the diagnosis of Brucella ovis infection. J Clin Microbiol. 1986 May;23(5):938–942. doi: 10.1128/jcm.23.5.938-942.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ris D. R. The persistence of antibodies against Brucella ovis and Brucella abortus in rams following vaccination: a field study. N Z Vet J. 1967 Jun;15(6):94–98. doi: 10.1080/00480169.1967.33702. [DOI] [PubMed] [Google Scholar]
  33. Roop R. M., 2nd, Jeffers G., Bagchi T., Walker J., Enright F. M., Schurig G. G. Experimental infection of goat fetuses in utero with a stable, rough mutant of Brucella abortus. Res Vet Sci. 1991 Sep;51(2):123–127. doi: 10.1016/0034-5288(91)90001-5. [DOI] [PubMed] [Google Scholar]
  34. Samartino L. E., Enright F. M. Interaction of bovine chorioallantoic membrane explants with three strains of Brucella abortus. Am J Vet Res. 1992 Mar;53(3):359–363. [PubMed] [Google Scholar]
  35. Schurig G. G., Hammerberg C., Finkler B. R. Monoclonal antibodies to Brucella surface antigens associated with the smooth lipopolysaccharide complex. Am J Vet Res. 1984 May;45(5):967–971. [PubMed] [Google Scholar]
  36. Schurig G. G., Roop R. M., 2nd, Bagchi T., Boyle S., Buhrman D., Sriranganathan N. Biological properties of RB51; a stable rough strain of Brucella abortus. Vet Microbiol. 1991 Jul;28(2):171–188. doi: 10.1016/0378-1135(91)90091-s. [DOI] [PubMed] [Google Scholar]
  37. Swift B. L., Maki L. R. Immunologic studies on three ram epididymitis bacterins. Cornell Vet. 1968 Oct;48(4):659–665. [PubMed] [Google Scholar]
  38. Tabatabai L. B., Deyoe B. L., Ritchie A. E. Isolation and characterization of toxic fractions from Brucella abortus. Infect Immun. 1979 Nov;26(2):668–679. doi: 10.1128/iai.26.2.668-679.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tobias L., Schurig G. G., Cordes D. O. Comparative behaviour of Brucella abortus strains 19 and RB51 in the pregnant mouse. Res Vet Sci. 1992 Sep;53(2):179–183. doi: 10.1016/0034-5288(92)90107-d. [DOI] [PubMed] [Google Scholar]
  40. Tsang V. C., Wilson B. C., Maddison S. E. Kinetic studies of a quantitative single-tube enzyme-linked immunosorbent assay. Clin Chem. 1980 Aug;26(9):1255–1260. [PubMed] [Google Scholar]
  41. Winter A. J., Rowe G. E., Duncan J. R., Eis M. J., Widom J., Ganem B., Morein B. Effectiveness of natural and synthetic complexes of porin and O polysaccharide as vaccines against Brucella abortus in mice. Infect Immun. 1988 Nov;56(11):2808–2817. doi: 10.1128/iai.56.11.2808-2817.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Young E. J. Human brucellosis. Rev Infect Dis. 1983 Sep-Oct;5(5):821–842. doi: 10.1093/clinids/5.5.821. [DOI] [PubMed] [Google Scholar]

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