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Infection and Immunity, June 2002, p. 2926-2932, Vol. 70, No. 6
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.6.2926-2932.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification by Mass Spectrometry of CD8⁺-T-Cell Mycobacterium tuberculosis Epitopes within the Rv0341 Gene Product

David C. Flyer,^1, Venkatesh Ramakrishna,^1* Cara Miller,^1, Helen Myers,¹ Melanie McDaniel,¹ Karen Root,² Caroline Flournoy,³ Victor H. Engelhard,³ David H. Canaday,⁴ Jarrod A. Marto,^2, Mark M. Ross,^1, Donald F. Hunt,² Jeffrey Shabanowitz,² and Forest M. White²

Argonex, Inc., Charlottesville, Virginia 22903,¹ Department of Chemistry,² Beirne Carter Center for Microbiology and Immunology, University of Virginia, Charlottesville, Virginia 22904,³ Division of Infectious Diseases, Case Western Reserve University, Cleveland, Ohio 44106⁴

Received 6 November 2001/ Returned for modification 21 December 2001/ Accepted 19 March 2002

Identification of Mycobacterium tuberculosis proteins that can provide immunological protection against tuberculosis is essential for the development of a more effective vaccine. To identify new vaccine targets, we have used immunoaffinity chromatography to isolate class I HLA-A*0201-peptide complexes from M. tuberculosis-infected cells and sequenced the isolated peptides by mass spectrometry. From this material, we have identified three peptides derived from a single M. tuberculosis protein that is encoded by the M. tuberculosis Rv0341 gene. Although no known protein encoded by the Rv0341 gene has been described, it is predicted to give rise to a 479-amino-acid protein with a molecular mass of 43.9 kDa. The three peptides identified are all nested and were found to be antigenic, in that they were capable of inducing peptide-specific, CD8⁺ T cells from healthy blood donors in vitro and capable of recognizing and lysing M. tuberculosis-infected dendritic cells. This methodology provides a powerful tool for the identification of M. tuberculosis proteins that can be evaluated as potential vaccine candidates.

Editor: S. H. E. Kaufmann

Present address: IOMAI Corp., Gaithersburg, MD 20878.

Present address: Wyeth-Lederle Viral Vaccine Immunology, Pearl River, NY 10956.

Present address: MDS Proteomics, Charlottesville, VA 22904.

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