Additional file 2: of Antigen-specific CD8+ memory stem T cells generated from human peripheral blood effectively eradicate allogeneic targets in mice

Figure S2. Our TSCM preparation strategy can be used to generate a single epitope-specific TSCM cells. A Procedure for preparation of AFP-specific, allogeneic TSCM. T2 cells express only empty HLA-A2 allele and no other HLA allele. Alpha fetoprotein (AFP) is a tumor associated antigen of hepatocarcinoma, the hAFP158–166 (FMNKFIYEI) is an HLA-A2 restricted peptide. When pulsed with the AFP peptide, T2 cells were able to present the AFP/HLA-A2 complex. The AFP-specific TSCM were raised by co-culturing HLA-A2 negative (HLA-A2-ve) PBLs and the T2 cells pulsed with the AFP peptide (T2/AFP). In a procedure of an allogeneic co-culture, differentiation inhibition, proliferation sorting and cytokine expansion, the AFP/HLA-A2 complex-specific TSCM cells were produced. B Co-culture by mixing of 1 × 107 PBLs and 2 × 106 T2/AFP on day 0, a prolonged cytokine expansion was required to generate 1 × 106 AFP-specific T cells. Data are represented as mean ± SD of four individual experiments. C–E The prepared TSCM cells were AFP-specific. The prepared TSCM cells were incubated with the T2/AFP and T2 cells pulsed with an irrelevant peptide HBcAg18–27 (T2/HBC), respectively. After 4 h incubation, the T cell subsets and their intracellular IFN-γ production were detected. Representative FCM plots (C). TSCM cells differentiated more when incubated with the T2/AFP (D). The daughter cells showed more frequent IFN-γ positive cells when incubated with the T2/AFP (E). Data are represented as mean ± SD of four individual experiments (** p < 0.01). (JPG 404 kb