FIGURE 1.

MUC1 CAR expression in transduced murine T cells. A, Schematic of murine MUC1 CAR construct. TAB004 antibody derived ScFv was incorporated in CAR construct for targeting tumor form of human MUC1 (human tMUC1). B and C, MUC1 CAR expression on T cells. Mouse T cells were activated with anti-mouse CD3/CD28 beads for 3 days, followed by overnight CAR transduction with retrovirus supernatant. CAR expression was detected 4 days after transduction. B, Mouse T cells were enriched from naïve mouse spleens. C, Mouse CD8⁺ T cells were isolated from naïve mouse spleens. The CAR expression was determined by flow cytometry analysis of biotinylated-MUC1 core peptide binding with transduced T cells. Cells were gated on the CD8⁺ population. Dead cells were excluded by 7-AAD staining. CAR indicates chimeric antigen receptor; MUC1, mucin 1; tMUCI, tumor form of mucin 1.