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Infection and Immunity, April 2008, p. 1565-1571, Vol. 76, No. 4
0019-9567/08/$08.00+0     doi:10.1128/IAI.01381-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Identification of an HLA-A*0201-Restricted T-Cell Epitope on the MPT51 Protein, a Major Secreted Protein Derived from Mycobacterium tuberculosis, by MPT51 Overlapping Peptide Screening

Department of Infectious Diseases,¹ Department of Health Science,² Department of Internal Medicine, Second Division, Hamamatsu University School of Medicine, Hamamatsu 431-3192, Japan³

Received 15 October 2007/ Returned for modification 26 November 2007/ Accepted 28 December 2007

CD8⁺ T cells play a pivotal role in protection against Mycobacterium tuberculosis infection. We identified a novel HLA-A*0201-restricted CD8⁺ T-cell epitope on a dominant secreted antigen of M. tuberculosis, MPT51, in HLA-A*0201 transgenic HHD mice. HHD mice were immunized with plasmid DNA encoding MPT51 with gene gun bombardment, and gamma interferon (IFN-) production by the immune splenocytes was analyzed. In response to overlapping synthetic peptides covering the mature MPT51 sequence, the splenocytes were stimulated to produce IFN- by only one peptide, p51-70. Three-color flow cytometric analysis of intracellular IFN- and cell surface CD4 and CD8 staining revealed that the MPT51 p51-70 peptide contains an immunodominant CD8⁺ T-cell epitope. Further analysis using computer algorithms permitted identification of a bona fide T-cell epitope, p53-62. A major histocompatibility complex class I stabilization assay using T2 cells confirmed that this epitope binds to HLA-A*0201. The T cells were capable of lysing MPT51 p53-62 peptide-pulsed T2 cells. In addition, MPT51 p53-62-specific memory CD8⁺ T cells were found in tuberculin skin test-positive HLA-A*0201⁺ healthy individuals. Use of this HLA-A*0201-restricted CD8⁺ T-cell epitope for analysis of the role of MPT51-specific T cells in M. tuberculosis infection and for design of vaccines against tuberculosis is feasible.

Published ahead of print on 22 January 2008.

Editor: R. P. Morrison