Expression cloning of an immunodominant family of Mycobacterium tuberculosis antigens using human CD4+ T cells

Mark R. Alderson, Teresa Bement, Craig H. Day, Liqing Zhu, David Molesh, Yasir A.W. Skeiky, Rhea Coler, David M. Lewinsohn, Steven G. Reed, Davin C. Dillon

Research output: Contribution to journalArticle

97 Scopus citations

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 γ 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-γ 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.

Original languageEnglish (US)
Pages (from-to)551-559
Number of pages9
JournalJournal of Experimental Medicine
Volume191
Issue number3
DOIs
StatePublished - Feb 7 2000

Keywords

  • Antigen presentation
  • Expression cloning
  • Interferon γ
  • Intracellular pathogens
  • Mycobacterium tuberculosis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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