Extended Data Fig. 8: Spatial imaging identifies IL-2+ microniches that impact Th2 cells.
a-b, Representative images as in (a) and (b) showing CD45.1 co-staining with RNA targets Cd19, Prdm1, Gata3, Il2 and Il2ra. B cell zone and T cell area were annotated by Cd19 and Cd4 RNA regions. c, Zoom-in images show separately acquired regions of interest of the T-B border for data in Fig. 7f. d-h, Naïve CD45.1/2 + 1DER T cells were adoptive transferred into CD45.2 + IL2eGFP reporter recipients followed by HDM immunization for 3d. Distance (µm) of IL-2eGFP+ cells to annotated B cell zone (d, n = 4) and frequency of IL-2eGFP+ cells distribution across medLN (e, n = 4). f, number of IL-2eGFP+ cells from immune cells in the medLN after 2d HDM i.n. immunization by flow cytometry assay (n = 3). naive unimmunized IL-2eGFP mouse were set as control (n = 2). g, Distance of GATA3 + , GATA3 + IL-2Ra + , IL-2Ra + 1DER T cells (CD45.1+) within a 50-μm-radius circle from IL-2GFP+ cells (n = 4). h, abundance of indicated FoxP3+ and GATA3 + CD4 + T cells surrounding IL2GFP+ cells within 50-μm-radius circle (n = 4). Data are shown as means ± SD above and representative of two independent experiments. Kruskal-Wallis one-way ANOVA test (g), Ordinary two-way ANOVA test (h). * P < 0.05, ** P < 0.01, ****P < 0.0001. Specific P values are as follows: g: CD45.1+IL2Ra+GATA3+ versus CD45.1+IL2Ra+=0.0098, CD45.1+GATA3+ versus CD45.1+IL2Ra+<0.0001; h: FoxP3+ (T cell zone) versus FoxP3+ (T-B border)=0.0333, ****P < 0.0001.
