Schematic representations. (A) Schematic representation of the pSIN second-generation lentiviral vector encoding the codon-optimized, murinized hybrid HLA-A0201–restricted pWT126-specific Cys-1 modified TCR α- and β-chain genes (or the HLA-A0201–restricted LMP2-specific Cys1 modified TCR chains) separated by a self-cleaving porcine tsechovirus 2A sequence. SFFV indicates spleen-forming focus virus; LTR, long terminal repeat; and WPRE, woodchuck hepatitis virus posttranscriptional regulatory element. (B) Schematic representation of the Cys-1 modified, codon-optimized hybrid TCR containing murine constant regions (black outline) and human variable regions. (C) Schematic representation of the hybrid HLA-A2Kb molecule expressed in HLA-A2Kb (A2Kb) transgenic mice. The hybrid MHC class I molecules in A2Kb mice contain human alpha1 and alpha2 domains that interact with the human HLA-A2–restricted TCR, fused to a murine alpha3 domain, which facilitates interaction with the murine CD8 coreceptor. (D) Schematic representation of TCR-transduced HSC transplantation. Lineage− BM stem cells (HSCs) were transduced with lentivirus at an MOI of 10. Each lethally irradiated recipient received 5 × 105 bulk transduced HSCs.
