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Infection and Immunity, May 2005, p. 2873-2880, Vol. 73, No. 5
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.5.2873-2880.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Brucella abortus Cu,Zn Superoxide Dismutase Is Required for Optimal Resistance to Oxidative Killing by Murine Macrophages and Wild-Type Virulence in Experimentally Infected Mice

Jason M. Gee,1,2 Michelle Wright Valderas,1,{dagger} Michael E. Kovach,2,{ddagger} Vanessa K. Grippe,2,¶ Gregory T. Robertson,3,# Wai-Leung Ng,3,% John M. Richardson,3 Malcolm E. Winkler,3,% and R. Martin Roop II1*

Department of Microbiology and Immunology, East Carolina University School of Medicine, Greenville, North Carolina 27834,1 Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130-3932,2 Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, Indiana 462853

Received 19 August 2004/ Returned for modification 16 November 2004/ Accepted 29 December 2004

Two-dimensional gel electrophoretic analysis of cell lysates from Brucella abortus 2308 and the isogenic hfq mutant Hfq3 revealed that the RNA binding protein Hfq (also known as host factor I or HF-I) is required for the optimal stationary phase production of the periplasmic Cu,Zn superoxide dismutase SodC. An isogenic sodC mutant, designated MEK2, was constructed from B. abortus 2308 by gene replacement, and the sodC mutant exhibited much greater susceptibility to killing by O2– generated by pyrogallol and the xanthine oxidase reaction than the parental 2308 strain supporting a role for SodC in protecting this bacterium from O2– of exogenous origin. The B. abortus sodC mutant was also found to be much more sensitive to killing by cultured resident peritoneal macrophages from C57BL6J mice than 2308, and the attenuation displayed by MEK2 in cultured murine macrophages was enhanced when these phagocytes were treated with gamma interferon (IFN-{gamma}). The attenuation displayed by the B. abortus sodC mutant in both resting and IFN-{gamma}-activated macrophages was alleviated, however, when these host cells were treated with the NADPH oxidase inhibitor apocynin. Consistent with its increased susceptibility to killing by cultured murine macrophages, the B. abortus sodC mutant also displayed significant attenuation in experimentally infected C57BL6J mice compared to the parental strain. These experimental findings indicate that SodC protects B. abortus 2308 from the respiratory burst of host macrophages. They also suggest that reduced SodC levels may contribute to the attenuation displayed by the B. abortus hfq mutant Hfq3 in the mouse model.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, East Carolina University School of Medicine, 600 Moye Boulevard, Greenville, NC 27834. Phone: (252) 744-1357. Fax: (252) 744-3535. E-mail: roopr{at}mail.ecu.edu.

Editor: V. J. DiRita

{dagger} Present address: Department of Veterinary Pathobiology, Oklahoma State University College of Veterinary Medicine, Stillwater, OK 74078.

{ddagger} Present address: Department of Biology, Baldwin-Wallace College, Berea, OH 44017.

Present address: Center for Biologics Evaluations and Research, Food and Drug Administration, Rockville, MD 20852.

# Present address: Cumbre, Inc., Dallas, TX 75235.

% Present address: Department of Biology, Indiana University, Bloomington, IN 47405.

Infection and Immunity, May 2005, p. 2873-2880, Vol. 73, No. 5
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.5.2873-2880.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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