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Infection and Immunity, February 2003, p. 621-628, Vol. 71, No. 2
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.2.621-628.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Nramp1 Is Not a Major Determinant in the Control of Brucella melitensis Infection in Mice

Laurence A. Guilloteau,1* Jacques Dornand,2 Antoine Gross,2 Michel Olivier,1 Fabienne Cortade,1 Yves Le Vern,3 and Dominique Kerboeuf3

Pathologie Infectieuse et Immunologie,1 Service commun de cytométrie en flux, Institut National de la Recherche Agronomique, Nouzilly,3 Institut National de la Santé et de la Recherche Médicale U-431, Montpellier, France2

Received 8 July 2002/ Returned for modification 3 September 2002/ Accepted 1 November 2002

Brucella, the causative agent of brucellosis in animals and humans, can survive and proliferate within macrophages. Macrophages mediate mouse resistance to various pathogens through the expression of the Nramp1 gene. The role of this gene in the control of Brucella infection was investigated. When BALB/c mice (Nramp1s) and C.CB congenic mice (Nramp1r) were infected with Brucella melitensis, the number of Brucella organisms per spleen was significantly larger in the C.CB mice than in the BALB/c mice during the first week postinfection (p.i.). This Nramp1-linked susceptibility to Brucella was temporary, since similar numbers of Brucella were recovered from the two strains of mice 2 weeks p.i. The effect of Nramp1 expression occurred within splenocytes intracellularly infected by Brucella. However, there was no difference between in vitro replication rates of Brucella in macrophages isolated from the two strains of mice infected in vivo or in Nramp1 RAW264 transfectants. In mice, infection with Brucella induced an inflammatory response, resulting in splenomegaly and recruitment of phagocytes in the spleen, which was amplified in C.CB mice. Reverse transcription-PCR (RT-PCR), performed 5 days p.i., showed that inducible nitric oxide synthase, tumor necrosis factor alpha (TNF-{alpha}), interleukin-12 p40 (IL-12p40), gamma interferon (IFN-{gamma}), and IL-10 mRNAs were similarly induced in spleens of the two strains. In contrast, the mRNA of KC, a C-X-C chemokine, was induced only in infected C.CB mice at this time. This pattern of mRNA expression was maintained at 14 days p.i., with IFN-{gamma} and IL-12p40 mRNAs being more intensively induced in the infected C.CB mice, but TNF-{alpha} mRNA was no longer induced. The higher recruitment of neutrophils observed in the spleens of infected C.CB mice could explain the temporary susceptibility of C.CB mice to B. melitensis infection. In contrast to infections with Salmonella, Leishmania, and Mycobacterium, the expression of the Nramp1 gene appears to be of limited importance for the natural resistance of mice to Brucella.

* Corresponding author. Mailing address: Unité de Pathologie Infectieuse et Immunologie, INRA, 37380 Nouzilly, France. Phone: (33) 2 47 42 78 90. Fax: (33) 2 47 42 77 79. E-mail: Laurence.Guilloteau{at}tours.inra.fr.

Editor: D. L. Burns

Infection and Immunity, February 2003, p. 621-628, Vol. 71, No. 2
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.2.621-628.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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