| Vector | lentivirus |
| Construct | scFv-4-1BB-CD3ζ |
| Transgene/Inserted gene | anti-CD19 single-chain variable fragment; 4-1BB costimulatory domain |
| Vector production method | CD4+ and CD8+ T cells were picked out from PBMCs, which were acquired from patients by leukapheresis, with the use of magnetic beads (Miltenyi Biotec). The two kinds of T cells were separately cultured in bottles pre-coated with CD3 and CD28, with the existence of IL-2 and IFN-γ. They were transduced to CAR-T cells with lentivirus vectors encoding anti-CD19 single-chain variable fragment, 4-1BB costimulatory domain, etc. during the following 5 days. Next, the virus vectors would be washed away and tests to ensure effectivity and safety of CAR-T cells would be carried out. When desirable amounts of CD4+ and CD8+ CAR-T cells were attained, a mixture of them would be formed in a 1:1 ratio. |
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