细胞过继免疫治疗肿瘤的临床研究进展__培训课件.ppt

* * FIGURE 2. Gammaretroviral vector designs. Shown are examples of gammaretroviral vector designs that have been used in the Surgery Branch, National Cancer Institute, to treat cancer patients with genetically modified T cells. TCR vectors (top) require expression of 2 proteins (the alpha and beta chains of the TCR), which can be done by the use of an internal promoter (such as PGK), an internal ribosome entry site (IRES), or a picornavirus ribosomal skip peptide (2A). CAR vectors (bottom) express an antitumor antigen single chain antibody (scFv) linked to T-cell signaling domains. Second-generation CAR vectors generally use a combinations of CD28 plus CD3zeta signaling domains, whereas third-generation CAR vectors include additional elements such as a hinge and transmembrane domain from CD8 and the second costimulatory elements, eg, derived from the 4-1BB gene. The specific proteins that have been targeted in Surgery Branch clinical trials using these vector designs is as indicated on the right of the figure. LTR indicates long terminal repeat; sd, RNA splice donor; sa, RNA splice acceptor; , packaging signal; CEA, carcinoembryonic antigen; and arrows, direction of transcription. * * * * * * * Whole blood (70 ml) was collected in BD Vacutainer blood collection tubes with sodium heparin and directly centrifuged to isolate PBMCs. PBMCs were stimulated with 5-mM zoledronic acid (Novartis, Basel, Switzerland) in AlyS203-gd medium (Cell Science and Technology Institute, Sendai, Japan) containing 1000 IU ml1 human recombinant IL-2 and 10% autologous serum at the beginning of the culture. * * Schemes for adoptive transfer of autologous, vaccine-primed, in vitro–expanded T cells. Patients are primed with tumor vaccine followed by lymphocyte harvest. Autologous T cells are harvested from peripheral blood (i) or draining lymph nodes (ii), undergo polyclonal in vitro activation and expansion, and are reinfused after lymphodepleting chemotherapy. Antigen-specific immune fun