Supplementary Figure S11 from Overexpression of an Engineered SERPINB9 Enhances Allogeneic T-cell Persistence and Efficacy

Supplementary Figure S11. Overexpression of SB9(CAS) in activated CD30.CAR with 4-1BB spacer (CD30.bb) TCRαβKO ATCs and CD19.CAR TCRαβKO ATCs protects them from allogeneic rejection and enhances anti-tumor efficacy in-vitro as described in Figure 3A. A, Flow cytometry analysis of NALM6 cells modified to overexpress truncated CD30 and were gene-edited for the knockout of HLA class I and II (HLADKO NALM6-CD30). HLADKO NALM6-CD30 cells were purified either by FACS or clonal selection. B and C, Representative ‘donor’ CD30.bb TCRαβKO ATC (B) or CD19.CAR TCRαβKO ATC (C) donor cell death (%) calculated by (donor cell count(co-culture of donor and tumor)- donor cell count(tri-culture))/(donor cell count (co-culture of donor and tumor))*100%, over 3 days in tri-culture. The assay was performed in the absence of cytokines. D and E, The tumor cell numbers counted by flow cytometry analysis on day 3 of tri-culture with ‘donor’ CD30.bb TCRαβKO ATCs (D) or CD19.CAR TCRαβKO ATCs (E) and respective ‘recipient’ Allo-T cells. The bar chart represents the median ± SD (n = 2).