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Infection and Immunity, July 2004, p. 3829-3837, Vol. 72, No. 7
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.7.3829-3837.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Identification of Murine H2-D^d- and H2-A^b-Restricted T-Cell Epitopes on a Novel Protective Antigen, MPT51, of Mycobacterium tuberculosis

Mina Suzuki, Taiki Aoshi, Toshi Nagata, and Yukio Koide^*

Department of Microbiology and Immunology, Hamamatsu University School of Medicine, Hamamatsu 431-3192, Japan

Received 15 September 2003/ Returned for modification 11 December 2003/ Accepted 9 March 2004

Both CD4⁺ type 1 helper T (Th1) cells and CD8⁺ cytotoxic T lymphocytes (CTL) play pivotal roles in protection against Mycobacterium tuberculosis infection. Here, we identified Th1 and CTL epitopes on a novel protective antigen, MPT51, in BALB/c and C57BL/6 mice. Mice were immunized with plasmid DNA encoding MPT51 by using a gene gun, and gamma interferon (IFN-) production from the immune spleen cells was analyzed in response to a synthetic overlapping peptide library covering the mature MPT51 sequence. In BALB/c mice, only one peptide, p21-40, appeared to stimulate the immune splenocytes to produce IFN-. Flow cytometric analysis with intracellular IFN- and the T-cell phenotype revealed that the p21-40 peptide contains an immunodominant CD8⁺ T-cell epitope. Further analysis with a computer-assisted algorithm permitted identification of a T-cell epitope, p24-32. In addition, a major histocompatibility complex class I stabilization assay with TAP2-deficient RMA-S cells transfected with K^d, D^d, or L^d indicated that the epitope is presented by D^d. Finally, we proved that the p24-32/D^d complex is recognized by IFN--producing CTL. In C57BL/6 mice, we observed H2-A^b-restricted dominant and subdominant Th1 epitopes by using T-cell subset depletion analysis and three-color flow cytometry. The data obtained are useful for analyzing the role of MPT51-specific T cells in protective immunity and for designing a vaccine against M. tuberculosis infection.

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* Corresponding author. Mailing address: Department of Microbiology and Immunology, Hamamatsu University School of Medicine, 1-20-1 Handa-yama, Hamamatsu 431-3192, Japan. Phone: 81-53-435-2334. Fax: 81-53-435-2335. E-mail: koidelb{at}hama-med.ac.jp.

Editor: S. H. E. Kaufmann

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Infection and Immunity, July 2004, p. 3829-3837, Vol. 72, No. 7
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.7.3829-3837.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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• Aoshi, T., Nagata, T., Suzuki, M., Uchijima, M., Hashimoto, D., Rafiei, A., Suda, T., Chida, K., Koide, Y. (2008). 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. Infect. Immun. 76: 1565-1571 [Abstract] [Full Text]