Extended Data Fig. 10: Differential expression kinetics of IL-1R1 in human cells, murine cells and a humanized mouse model.

(a) Overview of stimulation experiments for murine (left) and human (right) purified Treg populations. (b) Murine thymic γδ T cells (lower plot) were used as a positive control to validate the anti-mouse IL-1R1-PE antibody signal. (c) Representative plots depicting IL-1R1 and CD69 expression on unstimulated (black) and TCR stimulated (red) Tregs after 48 h. (d) Quantification of IL-1R1 expression after stimulation for 1 or 2 days in murine (top panel, n = 3) and human Tregs (bottom panel, n = 5), highlighting the discrepancy in expression level and kinetics. (e) Overview and timeline of human SCC15-tumor experiment in humanized MISTRG mice. Bottom, photograph of 5 tumors after collection. (f) Representative flow cytometry plots showing similar expression patterns for a set of key T cell markers in primary human HNSCC biopsies (top) and SCC15 tumors in MISTRG mice reconstituted with human immune cells (bottom). (g) Representative plots and quantification showing a similar increase in Treg frequencies between human blood/HNSCC tumor tissue (top) and humanized mouse blood/SCC15 tumor tissue (bottom). n = 6 for the humanized mouse samples, and n = 14 for the human HNSCC samples. (h) Representative plots and quantification showing that IL-1R1 expression is detectable, but under-represented in the humanized mouse model (n = 5) compared to primary human HNSCC tissue (see main Figure 3c). Error bars represent mean +/- SD. Statistical analyses were performed using one-way ANOVA with Tukey’s multiple comparisons test for (h) or using a two-tailed paired t-test for (g).