Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Jun;64(6):2188–2192. doi: 10.1128/iai.64.6.2188-2192.1996

Deletion of purE attenuates Brucella melitensis infection in mice.

R M Crawford 1, L Van De Verg 1, L Yuan 1, T L Hadfield 1, R L Warren 1, E S Drazek 1, H H Houng 1, C Hammack 1, K Sasala 1, T Polsinelli 1, J Thompson 1, D L Hoover 1
PMCID: PMC174054  PMID: 8675325

Abstract

We previously showed that a purE mutant (delta purE201) of Brucella melitensis 16M is attenuated for growth in cultured human monocytes (E. S. Drazek, H. H. Houng, R. M. Crawford, T. L. Hadfield, D. L. Hoover, and R. L. Warren, Infect. Immun. 63:3297-3301, 1995). To determine if this strain is attenuated in animals, we compared the growth of the delta purE201 mutant with that of strain 16M in BALB/c mice. The number of bacteria in the spleen and spleen weight peaked for both strains between 1 and 2 weeks postinfection (p.i.), though the number of delta purE201 cells was significantly less than the number of 16M cells recovered from the spleens of infected mice. During the next 6 weeks, delta purE201 was essentially eliminated from infected mice (three of five mice sterile; < 100 CFU in two of live mice at 8 weeks p.i.), whereas bacteria persisted at a high level in the spleens of 16M-infected mice (about 106 CFU per spleen). The number of bacteria in the livers and lungs of mice infected with either strain paralleled those in the spleen. Mice infected with 16M had a strong inflammatory response, developing dramatic and prolonged splenomegaly (five to eight times normal spleen weight) and producing serum interleukin-6. In contrast, mice infected with delta purE201 developed only mild, transient splenomegaly at 1 week p.i. and produced no interleukin-6 in their serum. We further characterized the host response to infection by measuring changes in immune spleen cell populations by flow cytometry. CD4- and CD8-positive lymphocytes declined by I week in both experimental groups, while MAC-1-positive cells increased. T-cell subpopulations remained low or declined further, and MAC-1 cells increased to three times normal levels during 8 weeks of infection with 16M but returned to normal by 4 weeks after infection with delta purE201. These results document infectivity and attenuation of delta purE201 and suggest that it should be further evaluated as a potential vaccine.

Full Text

The Full Text of this article is available as a PDF (263.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. 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]
  2. 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]
  3. Drazek E. S., Houng H. S., Crawford R. M., Hadfield T. L., Hoover D. L., Warren R. L. Deletion of purE attenuates Brucella melitensis 16M for growth in human monocyte-derived macrophages. Infect Immun. 1995 Sep;63(9):3297–3301. doi: 10.1128/iai.63.9.3297-3301.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Helle M., Boeije L., Aarden L. A. Functional discrimination between interleukin 6 and interleukin 1. Eur J Immunol. 1988 Oct;18(10):1535–1540. doi: 10.1002/eji.1830181010. [DOI] [PubMed] [Google Scholar]
  6. Hopkins S. J., Humphreys M. Simple, sensitive and specific bioassay of interleukin-1. J Immunol Methods. 1989 Jun 21;120(2):271–276. doi: 10.1016/0022-1759(89)90252-4. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. McFarland W. C., Stocker B. A. Effect of different purine auxotrophic mutations on mouse-virulence of a Vi-positive strain of Salmonella dublin and of two strains of Salmonella typhimurium. Microb Pathog. 1987 Aug;3(2):129–141. doi: 10.1016/0882-4010(87)90071-4. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Nicoletti P. Vaccination against Brucella. Adv Biotechnol Processes. 1990;13:147–168. [PubMed] [Google Scholar]
  11. Pugh G. W., Jr, Zehr E. S., Meador V. P., Phillips M., McDonald T. J., Deyoe B. L. Immunologic, histopathologic, and bacteriologic responses of five strains of mice to Brucella abortus strain 2308. Am J Vet Res. 1989 Mar;50(3):323–328. [PubMed] [Google Scholar]
  12. Sangari F. J., García-Lobo J. M., Agüero J. The Brucella abortus vaccine strain B19 carries a deletion in the erythritol catabolic genes. FEMS Microbiol Lett. 1994 Sep 1;121(3):337–342. doi: 10.1111/j.1574-6968.1994.tb07123.x. [DOI] [PubMed] [Google Scholar]
  13. Stevens M. G., Olsen S. C., Cheville N. F. Lymphocyte proliferation in response to immunodominant antigens of Brucella abortus 2308 and RB51 in strain 2308-infected cattle. Infect Immun. 1994 Oct;62(10):4646–4649. doi: 10.1128/iai.62.10.4646-4649.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Stevens M. G., Olsen S. C., Pugh G. W., Jr, Brees D. Comparison of immune responses and resistance to brucellosis in mice vaccinated with Brucella abortus 19 or RB51. Infect Immun. 1995 Jan;63(1):264–270. doi: 10.1128/iai.63.1.264-270.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Zhan Y., Cheers C. Endogenous gamma interferon mediates resistance to Brucella abortus infection. Infect Immun. 1993 Nov;61(11):4899–4901. doi: 10.1128/iai.61.11.4899-4901.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Zhan Y., Kelso A., Cheers C. Cytokine production in the murine response to brucella infection or immunization with antigenic extracts. Immunology. 1993 Nov;80(3):458–464. [PMC free article] [PubMed] [Google Scholar]
  17. Zhan Y., Yang J., Cheers C. Cytokine response of T-cell subsets from Brucella abortus-infected mice to soluble Brucella proteins. Infect Immun. 1993 Jul;61(7):2841–2847. doi: 10.1128/iai.61.7.2841-2847.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES