期刊
JOURNAL OF EXPERIMENTAL MEDICINE
卷 191, 期 3, 页码 551-559出版社
ROCKEFELLER UNIV PRESS
DOI: 10.1084/jem.191.3.551
关键词
Mycobacterium tuberculosis; intracellular pathogens; antigen presentation; interferon gamma; expression cloning
资金
- NHLBI NIH HHS [T32 HL007287, 5T32HL07287] Funding Source: Medline
- NIAID NIH HHS [N01AI-25147] Funding Source: Medline
Development of a subunit vaccine for Mycobacterium tuberculosis (Mtb) is likely to be dependent on the identification of T cell antigens that induce strong proliferation and interferon gamma production from healthy purified protein derivative (PPD)(+) donors. We have developed a sensitive and rapid technique for screening an Mtb genomic library expressed in Escherichia coli using Mtb-specific CD4(+) T cells. Using this technique, we identified a family of highly related Mtb antigens. The gene of one family member encodes a 9.9-kD antigen, termed Mtb9.9A. Recombinant Mtb9.9A protein, expressed and purified from E. coli, elicited strong T cell proliferation and IFN-gamma production by peripheral blood mononuclear cells from PPD+ but not PPD- individuals. Southern blot analysis and examination of the Mtb genome sequence revealed a family of highly related genes. A T cell line from a PPD+ donor that failed to react with recombinant Mtb9.9A recognized one of the other family members, Mtb9.9C. Synthetic peptides were used to map the T cell epitope recognized by this line, and revealed a single amino acid substitution in this region when compared with Mtb9.9A. The direct identification of antigens using T cells from immune donors will undoubtedly be critical for the development of vaccines to several intracellular pathogens.
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