Figure 5.

Adoptive transfer of BW6-specific CAR Tregs into Bw6⁺ NHP resulted in no adverse events

(A) Growth of Bw6-specific CAR Tregs from Bw6⁺ NHP in vitro. Each black line represents one independent expansion of Bw6-specific CAR Tregs from Bw6⁻ NHP, while red line represents growth of Bw6-specific CAR Tregs from Bw6⁺ NHP. Arrows indicate days of restimulation with irradiated Bw6.86 aAPCs.

(B) Expression of FoxP3 and Helios at the conclusion of manufacture of Bw6-specific CAR Tregs and CAR Teffs grown from Bw6⁺ animal.

(C) Bw6-specific CAR Tregs were co-cultured for 5 days with Carboxyfluorescein succinimidyl ester (CFSE)-labeled allogeneic PBMCs and α-CD3/α-CD28 beads at the indicated PBMC:Treg ratio to assess non-specific suppressor function as in Figure 3C. Histograms depict proliferation of the CD4⁻ CD8⁺ CFSE⁺ cells present in the allogenic PBMCs from Bw6⁻ animal.

(D) Following adoptive transfer of Bw6-specific CAR Tregs and HLA-A2-specific CAR Teffs to autologous Bw6⁺ recipient, whole blood was stained with HLA-A2 or HLA-B7 (Bw6⁺) tetramer at indicated time points. Dot plots are gated on CD4⁺ CD8⁻ FoxP3⁺ cells.