Journal: Journal for Immunotherapy of Cancer

Article Title: Induction of EBV latent membrane protein-2A (LMP2A)-specific T cells and construction of individualized TCR-engineered T cells for EBV-associated malignancies

doi: 10.1136/jitc-2021-002516

Figure Lengend Snippet: Phenotype and functional screening of S0 and S2 in each donor. (A) Flow cytometry analysis revealed percentages of CD4+ and CD8+ T cells. (B) Flow cytometry analysis revealed percentages of naive, effect, effect memory (TEM) and central memory T cells (TCM). (C) IFN-γ ELISA analysis of S2 T cells cocultured autologous DCs pulsed with LMP2A peptides. (D) IFN-γ ELISPOT analysis of S2 T cells cocultured autologous DCs pulsed with LMP2A peptides or not. Column histogram summarized the number of positive spots (E). The mean values from each group are plotted. Error bars represent the SEM (**p<0.01, ns, not significant, analyzed by one-way analysis of variance). NC, unstimulated peripheral T cells of each donor.

Article Snippet: Human IFN-γ ELISPOT Kit (Abcam, USA) was performed as the manufacturer’s protocols.

Techniques: Functional Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal: Journal for Immunotherapy of Cancer

Article Title: Induction of EBV latent membrane protein-2A (LMP2A)-specific T cells and construction of individualized TCR-engineered T cells for EBV-associated malignancies

doi: 10.1136/jitc-2021-002516

Figure Lengend Snippet: Functional assay of potential LMP2A-reactive TCR-Ts. (A) IFN-γ ELISA assay on TCR-Ts cocultured with DCs loaded with LMP2A peptides. (B) IFN-γ ELISPOT assay on TCR-Ts cocultured with DCs pulsed with LMP2A peptides. (C) CD25 and CD137 staining on TCR-Ts cocultured with DCs loaded with LMP2A peptides. Error bars represent the SEM (*p<0.05, **p<0.01, ***p<0.001, ns, not significant, analyzed by one-way analysis of variance). LMP2A, latent membrane protein-2A; NC, untransduced T cells.

Article Snippet: Human IFN-γ ELISPOT Kit (Abcam, USA) was performed as the manufacturer’s protocols.

Techniques: Functional Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Staining

Journal: Journal for Immunotherapy of Cancer

Article Title: Induction of EBV latent membrane protein-2A (LMP2A)-specific T cells and construction of individualized TCR-engineered T cells for EBV-associated malignancies

doi: 10.1136/jitc-2021-002516

Figure Lengend Snippet: Cytotoxic activity of TCR-Ts in vitro and in vivo. (A) IFN-γ ELISA assay and CD137 expression on TCR-Ts cocultured with LCL LMP2A . NC, untransduced T cells. (B) Cytolytic activity of TCR-Ts against corresponding LCL LMP2A at different E:T ratios. NC, untransduced T cells. (C) Schematic experimental plan. Each NOD/SCID mouse subcutaneously received 2×10 6 LCL LMP2A tumor cells on day 0. TCR-Ts or untransduced T cells (3×10 6 cells per mouse) were infused twice and IL-2 was administrated five times with a 2-day interval, followed by weekly tumor monitoring. (D) Antitumor activity of TCR-Ts against LCL LMP2A tumor cells in vivo. The tumor area is plotted on the y axis. Time after tumor cell injection is plotted on the x axis. The mean values from each group are plotted. Error bars represent the SEM (n=5 mice per group, *p<0.05, ***p<0.001, analyzed by one-way analysis of variance). NC, two intravenous injections of untransduced T cells and five intraperitoneal injection of IL-2 (1000 IU per mouse); TCR-T, two intravenous injections of TCR-Ts and five intraperitoneal injection of IL-2 (1000 IU per mouse). LCL, lymphoblastoid cell line; LMP2A, latent membrane protein-2A; NOD/SCID, non-obese diabetic severe combined immunodeficiency.

Article Snippet: Human IFN-γ ELISPOT Kit (Abcam, USA) was performed as the manufacturer’s protocols.

Techniques: Activity Assay, In Vitro, In Vivo, Enzyme-linked Immunosorbent Assay, Expressing, Injection