Generating AFP-specific TCR-T Cells

Production of AFP-specific TCR-T cells is similar to that described previously [33 (link)]. In brief, peripheral blood mononuclear cells (PBMCs) were isolated from healthy donors by density gradient centrifugation with Ficoll-hypaque. T cells were purified with T cell isolation kit (STEMCELL Technologies, Cambridge, MA) and stimulated with CD3/CD28 tetrameric antibody complex (STEMCELL Technologies, Cambridge, MA) for 24 h. Activated T cells were transduced with lentiviral particles containing the TCR gene. After 48 h, the expression of TCR was measured by flow cytometry and CD3/CD28 antibody complex was rinsed away. For TCR-T cells culture, RPMI-1640 medium containing 10% FBS and 50 IU/mL of interleukin-2 (IL-2) (Quanqi, Shandong, China) was refreshed every other day.