Figure 1.

Schematic illustration of representative TIL, TCR/CAR-T, and EBV-CTL manufacturing platforms. (a) Traditional and young TIL manufacturing scheme. Single-cell digests of resected tumor are plated in 24-well plates as microcultures. For traditional TILs, the microcultures are passaged as independent cultures. Screened and selected TIL cultures are further expanded with a 2-week rapid expansion procedure using OKT-3 antibody. For young TILs, microcultures are pooled without screening. The pooled cells undergo the same 2-week rapid expansion procedure to reach the target dose. During the whole process, cells are maintained in culture with 6000 IU mL⁻¹ of IL-2. (b) TCR/CAR-T manufacturing process. T cells are selected from washed apheresis product and activated by using CD3/28 Dynabeads and ClinExVivo magnetic particle concentrator (MPC). Activated T cells are transduced with TCR/CAR retroviral or lentiviral vectors, and transduced cells are expanded with WAVE bioreactor. CD3/28 magnetic Dynabeads are removed from the cells with MPC and end of the process cells are formulated for infusion. (c) CD8⁺ central memory TCR/CAR T-cell manufacturing process. PBMCs are first purified from apheresis product using Ficoll-Plaque gradient centrifugation, followed by CD4⁺, CD14⁺ and CD45RA⁺ cell depletion using anti-CD4, anti-CD14 and anti-CD45RA microbeads and CliniMACS. Collected cells undergo an additional CD62L positive selection procedure using anti-CD62L microbeads and CliniMACS. Selected CD8⁺CD62L⁺ cells are further activated with CD3/28 Dynabeads. Activated memory CD8⁺ cells are transduced with TCR/CAR vectors and expanded in vitro. Dynabeads are removed from the EOP cells using MCP before formulation. (d) Generation of multiviral antigen-specific T cells using G-Rex bioreactor. Donor PBMCs are pulsed with 15mer peptides mix spanning EBV, adenovirus, CMV, BK virus and human herpes virus antigen epitopes and transferred into G-Rex device. Multivirus antigen-specific T cells are expanded to high quantities in ~ 2 weeks of time in the presence of IL-4 and IL-7. CAR, chimeric antigen receptor; CTL, cytotoxic T lymphocytes; EBV, Epstein–Barr virus; IL, interleukin; PBMC, peripheral mononuclear blood cell; TCR, T-cell receptor; TIL, tumor-infiltrating lymphocyte.