Fig. 5

Superior proliferation and anti-HCC capacity of IL-21R-TCR-T. a The process of TCR-T and IL-21R-TCR-T receiving CD3/CD28 activation was shown. The cell number and TCR⁺ subset percentage were monitored every 4 days during the process. b The proliferating fold change of CD8⁺ TCR-T and IL-21R-TCR-T after CD3/CD28 activation was shown (n = 3). c The percentage of CD8⁺ TCR⁺ proportion in TCR-T and IL-21R-TCR-T during three rounds repeated coculture with HepG2 was shown (n = 3). d Representative pictures of HepG2 after 36 h coculture (Round 1, Round 2 and Round 3, the same procedure as Fig. 1a) with conventional TCR-T or IL-21R-TCR-T were shown. Scale bar = 100 µm. e The CTL activity of conventional TCR-T and IL-21R-TCR-T after 36 h coculture with HepG2 was measured by LDH assay (n = 4). f The supernatant IFN-γ levels of TCR-T or IL-21R-TCR-T during multiple round coculture with HepG2 were measured by ELISA (n = 4). g Schematics of the subcutaneous HepG2 model establishment and different TCR-T transferring in NPG mice. The tumor volume and TCR-T proportion in mice peripheral blood were monitored until the end of the experiment. h The mice tumor volume in each group after HepG2 implantation and mock-transduced T cells, TCR-T or IL-21R-TCR-T transfer was shown (n = 5). i The subcutaneous HepG2 tumor was isolated from NPG mice in each group at the end of the experiment, and the weight of isolated tumors was shown (n = 5). j The percentage of TCR⁺ T cells in total lymphocytes (mice and human lymphocytes) in the peripheral blood of each mouse 1, 7 and 14 days after TCR-T transfer was shown (n = 5). k NPG mice were subcutaneously injected with HepG2 and transferred with TCR-T or IL-21R-TCR-T. Mice were sacrificed 7 days after transferring and the cells in the tumor, peripheral blood, bone marrow and spleen were analyzed. l The numbers of tumor-infiltrating CD8⁺ TCR-T in each mouse were measured by flow cytometry and shown (n = 5). m The percentage of TCR⁺ T cells in tumor-infiltrating lymphocytes of each mouse 7 days after TCR-T transfer was shown (n = 5). n The TCR-T percentage in conventional TCR-T and IL-21R-TCR-T before and after 7 days of transfer was measured by flow cytometry. o The fold change of CD8⁺ TCR-T percentage in peripheral blood and tumor 7 days after transfer was shown (n = 5). Data were shown as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, NS not significant, ACT adoptive cell transfer