Supplementary Figure S12 from Overexpression of an Engineered SERPINB9 Enhances Allogeneic T-cell Persistence and Efficacy

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Supplementary Figure S12. Allogeneic rejection of ‘donor’ CD30.CAR with 4-1BB spacer (CD30.bb) EBVSTs occurs with the administration of alloreactive ‘recipient’ Allo-T cells in a CD30-positive B-cell acute lymphoblastic leukemia (B-ALL) mouse model. A, Schematic of in-vivo B-ALL allorejection model for (B-D), in which 2.5 x 106 of HLADKO NALM6-CD30.eGFP-ffLuc, FACS selected for truncated CD30 (tCD30) overexpression and HLA I and II knockout (HLADKO), were injected intravenously into NSG (MHCKO) mice. 15 days later, 5 x 106 ‘donor’ CD30.bb EBVSTs with or without 5 x 106 ‘recipient’ Allo-T cells which are CD30KO ATCs that were primed to recognize and kill ‘donor’ cells, were infused intravenously. B, Percentages of ‘donor’ CD30.bb EBVST cells in blood of individual mice, analyzed by flow cytometry at specified time points, (n=5 for CD30.bb + Allo-T treated group and n=6 for CD30.bb treated group). The data for CD30.bb + Allo-T group was derived from Fig. 3E, left. C, Bioluminescent images of HLADKO NALM6-CD30.eGFP-ffLuc tumor growth captured by IVIS Lumina S5 imaging system at specified time points. D, Quantified bioluminescent signals of tumor cells in individual mice at specified time points (top graph) and tumor burden on day 11 (bottom graph) normalized to the levels of disease on day 0, (n=5 for CD30.bb + Allo-T treated group and n=6 per treatment group for all other groups). Tumor bioluminescence data for no treatment and CD30.bb + Allo-T groups in (C) and (D) were derived from Fig. 3F and G. All graphs for in-vivo data denote mean + S.D. P values were determined using two-way ANOVA with Sidak’s correction for multiple comparisons (B) or one-way ANOVA with Dunnett’s T3 correction for multiple comparisons (D, bottom graph; day 11 post treatment).

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