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 Lewinsohn, Steven G. Reed, Davin C. Dillon

Research output: Contribution to journalArticle

94 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Organism Cloning
Mycobacterium tuberculosis
T-Lymphocytes
Proteins
Escherichia coli
Antigens
Subunit Vaccines
T-Lymphocyte Epitopes
Genomic Library
Viral Tumor Antigens
Amino Acid Substitution
Southern Blotting
Recombinant Proteins
Interferons
Genes
Blood Cells
Vaccines
Cell Proliferation
Mycobacterium tuberculosis antigens
Genome

Keywords

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

ASJC Scopus subject areas

  • Immunology

Cite this

Alderson, M. R., Bement, T., Day, C. H., Zhu, L., Molesh, D., Skeiky, Y. A. W., ... Dillon, D. C. (2000). Expression cloning of an immunodominant family of Mycobacterium tuberculosis antigens using human CD4+ T cells. Journal of Experimental Medicine, 191(3), 551-559. https://doi.org/10.1084/jem.191.3.551

Expression cloning of an immunodominant family of Mycobacterium tuberculosis antigens using human CD4+ T cells. / Alderson, Mark R.; Bement, Teresa; Day, Craig H.; Zhu, Liqing; Molesh, David; Skeiky, Yasir A W; Coler, Rhea; Lewinsohn, David; Reed, Steven G.; Dillon, Davin C.

In: Journal of Experimental Medicine, Vol. 191, No. 3, 07.02.2000, p. 551-559.

Research output: Contribution to journalArticle

Alderson, MR, Bement, T, Day, CH, Zhu, L, Molesh, D, Skeiky, YAW, Coler, R, Lewinsohn, D, Reed, SG & Dillon, DC 2000, 'Expression cloning of an immunodominant family of Mycobacterium tuberculosis antigens using human CD4+ T cells', Journal of Experimental Medicine, vol. 191, no. 3, pp. 551-559. https://doi.org/10.1084/jem.191.3.551
Alderson, Mark R. ; Bement, Teresa ; Day, Craig H. ; Zhu, Liqing ; Molesh, David ; Skeiky, Yasir A W ; Coler, Rhea ; Lewinsohn, David ; Reed, Steven G. ; Dillon, Davin C. / Expression cloning of an immunodominant family of Mycobacterium tuberculosis antigens using human CD4+ T cells. In: Journal of Experimental Medicine. 2000 ; Vol. 191, No. 3. pp. 551-559.
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