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. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: Cancer Res. 2014 Oct 8;74(23):6784–6795. doi: 10.1158/0008-5472.CAN-14-0043

Figure 2. Expression of phosphopeptide-specific murine TCR in human CD8 T-cells confers recognition of HLA-A2+ targets and effector function.

Figure 2

Human CD8 T-cells were electroporated with IVT RNA encoding phosphopeptide-specific murine TCR αβ chains, and assayed 12–14h later. (A, B) left panels, pIRS-2-specific (A) or pCDC25b-specific (B) human CD8 T-cells were co-cultured with the indicated targets for 5hrs and the percentage mTCRβ+ human CD8+ cells expressing surface either CD107a or intracellular IFN-γ or both (total responders) detected by flow cytometry. Right panels, pIRS-2-specific (A) or pCDC25b-specific (B) CD8 T-cells were cultured with phosphopeptide-pulsed (CFSEhi) or unpulsed (CFSElo) C1R-A2 targets, and specific cytotoxic activity was evaluated. (C) pIRS-2-specific or pCDC25b-specific human CD8 T-cells were co-cultured with the indicated human cancer cells and surface CD107a and/or intracellular IFN-γ on gated mTCRβ+ cells were detected by flow cytometry. Antigen expression was determined by Western blot and is shown in Figures 3 and 4. For all panels, data are representative of duplicate (triplicate for in vitro cytotoxicity assay) determinations in 2–5 experiments.