| Vector | lentivirus |
| Construct | scFv-CD28-4-1BB-CD3ζ |
| Vector type | NHP/TYF lentivector system |
| Transgene/Inserted gene | CD28 transmembrane and cytoplasmic domains; co-stimulatory 4-1BB intracellular TRAF binding domain; CD3z chain intracellular domain; inducible suicide caspase 9 gene |
| Viral genome modification | The DNA sequences of GD2 scFv were cloned into a pTYF lentiviral vector, followed by the lentiviral particle package. The GD2 CAR sequence was constructed with lentiviral 4SCAR. |
| Vector production method | Autologous GD2-specific 4SCAR-T cells were manufactured from 2 to 5 × 109 lymphocytes in the peripheral blood.Peripheral blood buffy coats were collected from patients and peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque plus (GE Healthcare). Lymphocytes were activated using phytohemagglutinin (5 µg/mL) for 2-3 days, after which the cells were maintained in TexMACS medium supplemented with human IL-2 (40 U/mL), IL-7 (20 U/mL), and IL-15 (10 U/mL). Activated T cells were transduced with GD2-specific 4SCAR vectors and expanded for six-ten days. |
| Additional feature | iCasp9 |