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. Author manuscript; available in PMC: 2017 Jul 8.
Published in final edited form as: Science. 2016 Jun 30;353(6295):179–184. doi: 10.1126/science.aaf6756

Fig. 4. Dsg3 CAAR-Tcells do not showoff-target toxicity.

Fig. 4

(A) Flow-cytometric quantification of CAAR-mediated signal transduction, as indicated by nuclear factor of activated Tcells (NFAT)–driven green fluorescent protein (GFP) expression in Jurkat reporter cells, using anti-Dsg3/1 Px44 CAR as a positive control for keratinocyte expression of Dsg3. Numbers indicate percentage of live, single cells in the GFP-positive gate. (B) Mean fluorescence intensity of GFP expression from (A). Representative data were replicated in at least five independent experiments with keratinocytes from different human donors. (C) 51Cr release after T cell–target cell coculture at indicated E:T ratios to measure cytotoxicity by Dsg3 CAAR-Ts and controls against human HaCat keratinocytes. Mean values of triplicate cultures are shown; similar results were obtained in two independent experiments. (D) Microscopic analysis of human skin xenografts to evaluate epidermal infiltration by Dsg3CAAR-Ts, positive control Px44 CAR-Ts, and negative control CART19. (Inset) Dyskeratotic keratinocytes surrounded by T cells. Dashed line shows dermal-epidermal junction. Scale bar, 125 μm. (E) Quantification of T cell infiltration into epidermis and dermis of human skin xenografts. Each symbol represents one mouse; ratio-paired t test, two-tailed, *P < 0.05; n.s. nonsignificant; data are pooled from two independent experiments.