Melanoma antigen-specific TCR-encoding γ-retroviral vectors, production of autologous TCR gene-modified lymphocytes, and treatment protocols. (A) Diagram of an MSCV-based γ-retroviral vector encoding the α-chain and β-chain of a MART-1–specific TCR (top) and of an MMLV-based γ-retroviral vector encoding the α-chain and β-chain of a gp100-specific TCR (bottom). (B) Timeline depicting production of autologous TCR gene-modified lymphocytes. The infusion product was composed of cells after short-term ex vivo culture (day 6-7) with or without a portion of cells exposed to an OKT-3-based rapid expansion protocol.²⁰  (C) Summary of TCR gene-modified lymphocyte protocols with administration of peptide vaccine (top) or fowlpox viral vaccine (bottom). All patients received nonmyeloablative lymphodepleting chemotherapy consisting of 2 days of cyclophosphamide (Cy) at 60 mg/kg followed by 5 days of fludarabine (Flu) at 25 mg/m².