Figure 2

MV-Edm improves antitumour immune responses of CD8⁺NKG2D⁺ cells against HCC cells. (a) LM3 or 97H cells were infected with MV-Edm (MOI = 1) for 24 h or were left uninfected, then cells were harvested and mixed with CD8⁺NKG2D⁺ cells at a ratio of 2:1 (E:T) for 12 h. The number of IFN-γ-producing CD8⁺NKG2D⁺ cells was determined by IFN-γ ELISPOT assay kit. Means + SD of triplicate from two independent experiments are shown. (b) LM3 and 97H cells were infected with MV-Edm (MOI = 1) for 24 h, then cells were stained by anti-MICA/B-PE before subjected to flow cytometry. Unstained cells were used as negative controls. An overlay of histograms representative of 3 independent experiments, and mean fluorescence intensity of MICA/B averaged from 3 independent experiments are shown. (c) LM3 and 97H cells were infected with MV-Edm at a MOI of 1 for 24 h, then CD8⁺NKG2D⁺ cells were added at a ratio of 2:1 (E:T) for another 24 h. CD8⁺NKG2D⁺ cells were then harvested and stained with CD3-FITC and FasL-PE before subjected to flow cytometry. Fluorescence intensity of FasL was determined in CD3⁺ cells. Unstained cells were used as negative controls. An overlay of histograms representative of 3 independent experiments, and the mean fluorescence intensity of FasL averaged from 3 independent experiments are shown. (d) LM3 and 97H cells were infected with MV-Edm (MOI = 1) for 24 and 48 h, then cells were stained by anti-Fas-FITC before subjected to flow cytometry. Unstained cells were used as negative controls. An overlay of histograms representative of 3 independent experiments, and mean fluorescence intensity of MICA/B averaged from 3 independent experiments are shown. ns, not significant, ** P < 0.01.