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Fig. 4 | BMC Research Notes

Fig. 4

From: Effector function of CTLs is increased by irradiated colorectal tumor cells that modulate OX-40L and 4-1BBL and is reversed following dual blockade

Fig. 4

Sensitivity to CEA- and MUC-specific T-cell mediated cytolysis in irradiated colorectal tumor cells. a HLA-A2 positive HCT116 and SW620 cells treated in vitro with 0 Gy (white bar), 5 Gy (gray bar) or 10 Gy (black bar) of ionizing radiation were used as targets in a 4 h CTL cytolysis assay. At 72 h post-IR, HLA-A2 restricted CEA-specific T cells were used as effector cells at an E:T of 25:1. b Irradiated (5 and 10 Gy) and non-irradiated (0 Gy) HCT116 and SW620 cells were used in a 4 h lysis assays with MUC-specific T cells. At 72 h post-IR, MUC-specific T cells were used as effector cells at an E:T of 25:1 (HCT116) or 12:1 (SW620). c HLA-A2 positive colo205 cells were used in a cytolysis assay with either CEA-specific or MUC-specific T cells at an E:T of 30:1. d HLA-A2 negative LS174T cells were used in a cytolysis assay with either CEA-specific or MUC-specific T cells at an E:T of 30:1.*P value <0.05. Error bars indicate variability in technical replicates. Experiments were repeated at least three times with similar results. e Irradiated (10 Gy) and non-irradiated (0 Gy) CRC cells were cultured and subsequently stained with PE-labeled antibodies for flow cytometry to measure surface expression of HLA-A2 and TAA proteins on the surface of colorectal tumor cells. Isotype control staining of irradiated cells was less than 5 % positive. Numbers indicate  % of cells positive and those in parenthesis indicate mean fluorescence intensity (MFI) of cells expressing molecule on the cell surface 72 h post-irradiation. (dashed line) indicates level of detection below background. CEA carcinoembryonic antigen, MUC Mucin-1

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