Fig. 2: TYRP1 CAR-T cell therapy optimized to detect the low levels of TYRP1 on the cell surface leads to cytotoxicity and cytokine release against patient-derived and murine melanoma models. | Nature Communications

Fig. 2: TYRP1 CAR-T cell therapy optimized to detect the low levels of TYRP1 on the cell surface leads to cytotoxicity and cytokine release against patient-derived and murine melanoma models.

From: CAR-T cell therapy targeting surface expression of TYRP1 to treat cutaneous and rare melanoma subtypes

Fig. 2

a Schematics of the 20D7SS-28ζ, 20D7SM-28ζ, and the 20D7SL-28ζ CAR-T cells. Graphical depictions were created with BioRender.com. b Human primary T cells transduced with the TYRP1 CARs. Representative flow cytometry plots showing CAR expression on the cell surface. CAR expression was detected with the Kip-1 anti-whitlow linker antibody. Representative flow cytometry showing transduction efficiency of one experiment out of two experiments performed using these constructs. c, d Co-culture of 20D7SS-28ζ, 20D7SM-28ζ, and the 20D7SL-28ζ with a panel of TYRP1high and TYRP1intermediate patient-derived cell lines. Mean ± SD (n = 4, independent co-cultures) are plotted. c Percentage of tumor cell growth inhibition normalized by the growth of the cell lines co-cultured with untransduced T cells at 96 h after co-culture at a 1:1 effector to target (E:T) ratio. d IFNγ released at 24 h after co-culture measured by ELISA (5:1, E:T ratio). e Cytotoxicity dose–response curves of 20D7SS-28ζ, 20D7SM-28ζ, and the 20D7SL-28ζ CAR-T cells upon 48 h co-culture with a panel of TYRP1high and TYRP1intermediate melanoma cell lines. Percentage of tumor cell growth inhibition normalized by the untransduced T cells control. Mean ± SD (n = 4, independent co-cultures) are plotted. f–h Antitumor activity of the 20D7SS-28ζ, 20D7SM-28ζ, and the 20D7SL-28ζ murine CAR-T cells upon co-culture with the B16 murine melanoma cell line. Untransduced murine T cells, or media alone are used as negative controls. Mean ± SD (n = 4, independent co-cultures) are plotted. f Time-course of B16 murine melanoma tumor cell growth and inhibition (5:1 E:T). g Cytotoxicity dose-response curves at 48 h after co-culture. h Dose–response IFNγ release at 24 h after co-culture. Cytokine secretion of T cells alone without stimulation is shown as a negative control. *p < 0.05 unpaired, two-tailed t test with Holm-Sidak adjustment for multiple comparisons. Source data and exact p values are provided as a Source Data file.

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