Fig. 5: Spatial transcriptomics of Pd1−/− Treg cell communication programs in TME.
From: PD-1 receptor deficiency enhances CD30+ Treg cell function in melanoma
a–g, Tumor-bearing Rag1−/− mice were reconstituted with different T cell subsets and then subjected to CosMx spatial transcriptomics: UMAPs of 9,231 cells derived from WT Teff/WT Treg cell and WT Teff/Pd1−/− Treg cell cohorts, color-coded by assigned cell type (left), and WT and Pd1−/− Treg cell cohorts decoupled (middle and right) (a); frequencies of all cells in the TME (b); immune cells subclusters and frequencies of immune cell subsets among WT and Pd1−/− Treg cells (c); a graphical representation of cell–cell interaction analysis (left) and numbers of cell–cell interactions of WT Treg cells (middle) and Pd1−/− Treg cells (right) (d); a graphical representation of SR and LR gene definition by manual annotation (left), and frequencies of LR and SR interactions of WT (middle) and Pd1−/− Treg cells (right) (e); and DC Il10 expression (f) and Gzmb expression in NK cells (g) according to clustered Wilcoxon rank-sum test are shown. h,i, Tamoxifen-treated, tumor-bearing Pd1fl/fl and Pd1fl/flFoxp3ERT2Cre mice were immunophenotyped on day 19: the frequencies of IL-10 expression in TIL DCs (n = 5 per group) (h) and GzmB expression in TIL NK cells (n = 10 for Pd1fl/fl and n = 9 for Pd1fl/flFoxp3ERT2Cre) (i) are shown. j, For mice as in a–g, FOVs showing colocalization of different WT and Pd1−/− Treg cell states and their interactions within the TME. k,l, Tamoxifen-treated, tumor-bearing Pd1fl/flFoxp3ERT2Cre mice were immunophenotyped (n = 8): representative flow cytometry results (k) and a summary of CD30 expression in Treg cell subsets (l) are shown. Data represent individual cells in f and g. In h, i and l, data are from three independent experiments, and the mean ± s.e.m. is shown. In h and i, each data point represents an individual mouse. In l, data points are matched within animals. Two-tailed clustered Wilcoxon rank-sum test was used for f and g, two-tailed unpaired Student’s t-test for h and i, and one-way ANOVA with Sidak’s multiple comparison test for l. *P < 0.05, **P < 0.01. Illustrations in d and e created using BioRender.com. ILC, innate lymphoid cell.
