Supplementary Figure S3 from Overexpression of an Engineered SERPINB9 Enhances Allogeneic T-cell Persistence and Efficacy

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Supplementary Figure S3. CD30.CAR with IgG1 spacer (CD30.Ig) TCRαβKO ATCs expressing different variants of SB9 had similar CD30.Ig expression and cytotoxicity. A, Expression of TCRαβ on CD3 wildtype (WT) T cells or T cells after TCRαβKO gene modification using Crispr gene editing measured by flow cytometry analysis. B, Expression of different variants of SB9 on CD30.Ig TCRαβKO ATCs measured by intracellular flow cytometry analysis of one representative donor. C, Relative median fluorescence intensity (MFI) of different variants of SB9, normalized to SB9 expression in CD30.Ig sample. The bars denote median ± SD (n = 3 donors). D, CD30.Ig expression on TCRαβKO ATCs expressing different variants of SB9, measured by flow cytometry analysis of a representative donor. E and F, Pooled data of CD30.Ig expression (%; E) and CD30.Ig expression (geometric mean fluorescence intensity, MFI; F) on CD30.Ig TCRαβKO ATCs, measured by flow cytometry analysis of n = 3 donors, where bars denote median ± SD. G, CD30.Ig TCRαβKO ATC-mediated cytolysis of KM-H2 cells, of a representative donor, measured using the xCELLigence system. CD30.Ig TCRαβKO ATCs manufactured from three donors were tested and found to have similar results.

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