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| Article Authors: | M R Alderson; T Bement; C H Day; L Zhu; D Molesh; Y A Skeiky; R Coler; D M Lewinsohn; S G Reed; D C Dillon |
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| Article Title: | Expression cloning of an immunodominant family of Mycobacterium tuberculosis antigens using human CD4(+) T cells. |
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| Reference ID: | 942 |
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| Abstract: | 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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| Affiliations: | Department of Immunology, Corixa Corporation, Seattle, Washington 98104, USA. alderson@corixa.com |
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| Date: | 2000 |
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| Reference Type: | Literature |
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| PubMed ID: | 10662800 |
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| Journal: | J Exp Med |
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| Journal Volume: | 191 |
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| Article Pages: | 551-60 |
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| Journal ISSN: | 1540-9538 |
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| Article Chemical List: | Antigens, Bacterial;Bacterial Vaccines;Mtb 9.9 antigen |
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| Article MeSH List: | Amino Acid Sequence; Antigens, Bacterial(chemistry; genetics; immunology); Bacterial Vaccines; CD4-Positive T-Lymphocytes(immunology; metabolism; microbiology); Cell Line; Cloning, Molecular; Escherichia coli(genetics; metabolism); Genes, Bacterial; Genomic Library; Humans; Molecular Sequence Data; Mycobacterium tuberculosis(immunology) |
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| Curation Last Updated: | 2013-05-28 20:04:57 |
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