SING: a novel strategy for identifying tumor-specific, CTL-recognized tumor antigens.

Tong Zhang, Xianghui He, Tom C. Tsang, David T. Harris

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Traditional methods for identifying T cell-recognized tumor antigens (Ags) are laborious and time-consuming. In an attempt to simplify the procedure, a novel strategy, SING (SIgnal transduction molecule-mediated, NFAT-controlled, GFP expression) was established as a direct approach for cloning T cell-recognized tumor Ags. In the SING system, a mouse T cell line (BW5147) was transduced with a chimeric H-2Kb construct containing T cell-signaling domains and a green fluorescent protein (GFP) reporter gene under the transcriptional control of nuclear factor of activated T cells (NFAT). The resultant BW5147 cells were named BS cells. This cell line could "sense" TCR stimulation through the T cell-signaling domains after coculture with Ag-specific T cells and then become fluorescent (expressing green fluorescence protein, GFP+) in the presence of Ag peptides. The interaction between BS cells and Ag-specific T cells could be enhanced by addition of costimulatory signals. Currently, BS cells have been optimized to "sense" TCR stimulation after being pulsed with the relevant peptides at concentrations as low as 10(-9) M. Endogenous Ag-expressing BS cells could also become fluorescent after coculture with Ag-specific T cells. Our results provide a proof of principle for using the SING system to directly isolate Ag-expressing BS cells from BS cell repertoires, which are retrovirally transduced with tumor-derived cDNA libraries. Once tumor Ag-marked BS cells are identified, the sequences encoding tumor Ags could be easily retrieved by PCR amplification of the genomic DNA using vector-specific primers.

Original languageEnglish (US)
Pages (from-to)600-602
Number of pages3
JournalFASEB Journal
Volume18
Issue number3
StatePublished - 2004

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T-cells
Neoplasm Antigens
Tumors
T-Lymphocytes
Neoplasms
Green Fluorescent Proteins
NFATC Transcription Factors
Cell signaling
Coculture Techniques
Cell Line
Signal transduction
Peptides
Cloning
Gene Library
Reporter Genes
Amplification
Organism Cloning
Signal Transduction
Genes
Fluorescence

Cite this

SING : a novel strategy for identifying tumor-specific, CTL-recognized tumor antigens. / Zhang, Tong; He, Xianghui; Tsang, Tom C.; Harris, David T.

In: FASEB Journal, Vol. 18, No. 3, 2004, p. 600-602.

Research output: Contribution to journalArticle

Zhang, Tong ; He, Xianghui ; Tsang, Tom C. ; Harris, David T. / SING : a novel strategy for identifying tumor-specific, CTL-recognized tumor antigens. In: FASEB Journal. 2004 ; Vol. 18, No. 3. pp. 600-602.
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