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Infection and Immunity, July 2006, p. 4002-4013, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00257-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification, Recombinant Expression, Immunolocalization in Macrophages, and T-Cell Responsiveness of the Major Extracellular Proteins of Francisella tularensis

Bai-Yu Lee, Marcus A. Horwitz, and Daniel L. Clemens*

Division of Infectious Diseases, Department of Medicine, Center for Health Sciences, University of California, Los Angeles, School of Medicine, Los Angeles, California 90095-1688

Received 16 February 2006/ Returned for modification 7 April 2006/ Accepted 24 April 2006

A safer and more effective vaccine than the previously developed live attenuated vaccine is needed for combating Francisella tularensis, a highly infectious bacterial pathogen. To search for potential candidates for inclusion in a new vaccine, we characterized the proteins present in the culture filtrates of a virulent recent clinical isolate and the attenuated live vaccine strain of F. tularensis using a proteomic approach. We identified a total of 12 proteins; among these, catalase-peroxidase was much more abundant in the culture filtrate of the virulent clinical isolate, whereas bacterioferritin was more abundant in the culture filtrate of the live vaccine strain. Streptolysin O treatment of infected human macrophages indicated that catalase-peroxidase and the heat shock protein GroEL are released intracellularly by actively growing F. tularensis. Mice immunized with F. tularensis developed significant cell-mediated immune responses to catalase-peroxidase, the heat shock protein GroEL, and bacterioferritin as measured by splenic lymphocyte proliferation and gamma interferon production. Finally, we expressed the major culture filtrate proteins that are promising vaccine candidates in Escherichia coli at high levels in soluble form to facilitate study of their immunobiology and potential role in vaccines.

* Corresponding author. Mailing address: Division of Infectious Diseases, 37-121 Center for the Health Sciences, University of California, Los Angeles, 10833 Le Conte Avenue, Los Angeles, CA 90095-1688. Phone: (310) 825-9324. Fax: (310) 793-7156. E-mail: dclemens{at}mednet.ucla.edu.

Editor: D. L. Burns

Infection and Immunity, July 2006, p. 4002-4013, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00257-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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