Fig. 1: Signal activation and gene expression profiles induced by orthogonal chimeric receptors.
From: Expanding the cytokine receptor alphabet reprograms T cells into diverse states
a, Schematic of orthogonal IL-2–IL-2Rβ pairs, whereby a mutant IL-2 interacts with a mutant IL-2Rβ that pairs with the common γc to trigger swappable ICD signalling. b, Natural or non-natural ICDs that pair with the γc. c–g, Orthogonal-receptor-expressing (YFP+) C57BL/6 WT CD3+ T cells were stimulated with MSA fusions of orthogonal mouse IL-2 (MSA–oIL-2) for 20 min. Cells were analysed for pSTAT. c, pSTAT3 signalling dose–response curves of o2R-, o10R-, o20R- and o22R-transduced WT T cells, plotted against the log10-transformed concentration of MSA–oIL-2. d, The relative pSTAT1, pSTAT3, pSTAT4, pSTAT5 and pSTAT6 mean fluorescence intensity (MFI) in T cells transduced with o2R, o4R, o7R, o9R and o21R treated with MSA–oIL-2 (10 µM), normalized to the YFP− controls. e, The relative pSTAT1, pSTAT3 and pSTAT5 MFI in T cells transduced with o2R, oIFNAR2, oIFNGR1 and oIFNLR1, treated with MSA–oIL-2 (10 µM), normalized to the YFP− controls. f, The relative pSTAT1, pSTAT3, pSTAT4 and pSTAT5 MFI in T cells transduced with o2R, o10R, o20R and o22R and treated with MSA–oIL-2 (10 µM), normalized to the YFP− controls. g, The relative pSTAT1, pSTAT3 and pSTAT5 MFI in T cells transduced with o2R, oEPOR and oGCSFR and treated with MSA–oIL-2 (10 µM), normalized to the YFP− controls. h–j, C57BL/6 WT T cells transduced with orthogonal receptors were stimulated with MSA–oIL-2 (5 μM) or recombinant cytokines (10 nM) for 6 h, and then analysed using RNA-seq. n = 3 biologically independent samples. h, PCA of RNA-seq data. i, The expression of STAT-driven gene signatures. FC, fold change compared with the no-ICD controls. j, Comparison of the STAT gene signatures of orthogonal chimera-transduced T cells treated with MSA–oIL-2 versus those treated with the indicated native cytokines. Data are mean ± s.e.m.
