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Infection and Immunity, May 2007, p. 2548-2561, Vol. 75, No. 5
0019-9567/07/$08.00+0     doi:10.1128/IAI.01974-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Proteomic and Immunoblot Analyses of Bartonella quintana Total Membrane Proteins Identify Antigens Recognized by Sera from Infected Patients

Jenni K. Boonjakuakul,^1, Helen L. Gerns,¹ Yu-Ting Chen,¹ Linda D. Hicks,² Michael F. Minnick,² Scott E. Dixon,³ Steven C. Hall,³ and Jane E. Koehler^1*

Division of Infectious Diseases, Department of Medicine, University of California at San Francisco, San Francisco, California 94143-0654,¹ Division of Biological Sciences, University of Montana, Missoula, Montana 59812,² Biomolecular Resource Center Mass Spectrometry Facility, University of California at San Francisco, San Francisco, California 94143³

Received 15 December 2006/ Returned for modification 21 January 2007/ Accepted 10 February 2007

Bartonella quintana is a fastidious, gram-negative, rod-shaped bacterium that causes prolonged bacteremia in immunocompetent humans and severe infections in immunocompromised individuals. We sought to define the outer membrane subproteome of B. quintana in order to obtain insight into the biology and pathogenesis of this emerging pathogen and to identify the predominant B. quintana antigens targeted by the human immune system during infection. We isolated the total membrane proteins of B. quintana and identified 60 proteins by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and peptide mass fingerprinting. Using the newly constructed proteome map, we then utilized two-dimensional immunoblotting with sera from 21 B. quintana-infected patients to identify 24 consistently recognized, immunoreactive B. quintana antigens that have potential relevance for pathogenesis and diagnosis. Among the outer membrane proteins, the variably expressed outer membrane protein adhesins (VompA and VompB), peptidyl-prolyl cis-trans-isomerase (PpI), and hemin-binding protein E (HbpE) were recognized most frequently by sera from patients, which is consistent with surface expression of these virulence factors during human infection.

Published ahead of print on 16 February 2007.

Editor: W. A. Petri, Jr.

Present address: Foodborne Contaminants Research Unit, Agricultural Research Service, U.S. Department of Agriculture, 800 Buchanan St., Albany, CA 94710.

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• MacKichan, J. K., Gerns, H. L., Chen, Y.-T., Zhang, P., Koehler, J. E. (2008). A SacB Mutagenesis Strategy Reveals that the Bartonella quintana Variably Expressed Outer Membrane Proteins Are Required for Bloodstream Infection of the Host. Infect. Immun. 76: 788-795 [Abstract] [Full Text]