Figure 3.

iPSC-β-Induced T Cell Activation and Killing Is Mediated by Direct T Cell-HLA Interaction

(A and B) PBMCs co-cultured for 48 h with autologous iPSC-β (n = 3 T1D donors, 3 differentiation batches per donor). iPSC-β were pre-treated with thap (5 μM for 5 h) and/or anti-HLA antibody for 30 min before co-culture.

(A) Flow cytometry after 48 h co-culture of T cell activation markers, CD25⁺ and CD69⁺, gated on CD8⁺ cells.

(B) Pro-inflammatory cytokine detection in supernatants collected after 48 h co-culture of PBMCs with autologous iPSC-β ± thap.

(C) Expression of CD25⁺ and CD69⁺, gated on CD8⁺ cells in a transwell system.

(D) Expression of the activation marker CD25⁺ and CD69⁺ on CD8⁺ cells after 48-h co-culture with autologous iPSC-β transduced with a lentivirus vector expressing a non-targeting (NT) or B2M guide RNA (gRNA) and Cas9 (n = 3 T1D donors, n = 3 differentiation batches per donor line). ^∗p < 0.05 and ^∗∗p < 0.01, Student’s t test. T1D¹, T1D², and T1D³ were pooled together. ns, non-significant.

(A–C) n = 3 T1D donors, n = 3 differentiation batches per donor line. T1D¹, T1D², and T1D³ were pooled together. ^∗p < 0.05, ^∗∗p < 0.005, and ^∗∗∗p < 0.0005; 2-way ANOVA.

See also Figure S3.