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Infection and Immunity, May 2005, p. 2665-2679, Vol. 73, No. 5
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.5.2665-2679.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Use of In Vivo-Induced Antigen Technology for Identification of Escherichia coli O157:H7 Proteins Expressed during Human Infection

Manohar John,^1,2, Indira T. Kudva,^1,2,* Robert W. Griffin,¹ Allen W. Dodson,¹ Bethany McManus,¹ Bryan Krastins,³ David Sarracino,³ Ann Progulske-Fox,⁴ Jeffrey D. Hillman,⁴ Martin Handfield,⁴ Phillip I. Tarr,⁵ and Stephen B. Calderwood^1,2,6

Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts 02114,¹ Department of Medicine, Harvard Medical School, Boston, Massachusetts 02115,² Harvard Partners Center For Genetics and Genomics, 65 Landsdowne Street, Cambridge, Massachusetts 02139,³ College of Dentistry, Department of Oral Biology, University of Florida, Gainesville, Florida 32610,⁴ Departments of Pediatrics and Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110,⁵ Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115⁶

Received 4 November 2004/ Returned for modification 14 December 2004/ Accepted 12 January 2005

Using in vivo-induced antigen technology (IVIAT), a modified immunoscreening technique that circumvents the need for animal models, we directly identified immunogenic Escherichia coli O157:H7 (O157) proteins expressed either specifically during human infection but not during growth under standard laboratory conditions or at significantly higher levels in vivo than in vitro. IVIAT identified 223 O157 proteins expressed during human infection, several of which were unique to this study. These in vivo-induced (ivi) proteins, encoded by ivi genes, mapped to the backbone, O islands (OIs), and pO157. Lack of in vitro expression of O157-specific ivi proteins was confirmed by proteomic analysis of a mid-exponential-phase culture of E. coli O157 grown in LB broth. Because ivi proteins are expressed in response to specific cues during infection and might help pathogens adapt to and counter hostile in vivo environments, those identified in this study are potential targets for drug and vaccine development. Also, such proteins may be exploited as markers of O157 infection in stool specimens.

Editor: A. D. O'Brien

Equal contribution.

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