| Vector | retrovirus |
| Construct | scFv-CD28-CD3ζ |
| Transgene/Inserted gene | HER2-specific murine scFv FRP5; CD28 transmembrane domain; CD28.ζ signaling domain |
| Vector production method | Peripheral-blood mononuclear cells were activated with immobilized CD3 antibody (Ortho Biotech, Toronto, Ontario, Canada) or immobilized CD3 and CD28 antibodies (P 6, 12, 13, 16, 17, 18; Miltenyi, Cologne, Germany) and recombinant IL-2 (100 U/mL; Proleukin, Chiron, Emeryville, CA) and then transduced on day 3 with retroviral particles encoding an HER2.CD28.ζ-CAR (Appendix, online only) in 24-well plates precoated with Retronectin (FN CH-296; Takara, Shiga, Japan). After transduction, T-cell lines were expanded in the presence of IL-2 (50 to 100 U/mL) added twice weekly until the specified cell dose was achieved. |
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