Figure 1.

Developing Different Generation EphA2-Specific CAR T Cells

(A) Scheme of EphA2 CARs. All CARs contained an N-terminal leader sequence, a codon-optimized synthetic gene encoding for human 4H5, a spacer region (16-amino acid IgG1 hinge), a CD28 or CD8α transmembrane domain, signaling domains derived from CD28, 41BB, and CD3.ζ, and tCD19, separated by 2A sequence. (B and C) Genetic modification of T cells was confirmed by FACS analysis using a CD19 antibody. Representative plots (B) and summary data (C) are shown (mean 74.1%–93.3%, n = 5–6 per CAR construct). Error bars represent mean with SD. (D) Expression of full-length EphA2-CARs by western blot analysis using a CD3-ζ antibody under denaturing and non-denaturing conditions.