Heterogeneity and Homogeneity in Treg responses to Interleukin-2 [scRNA-seq: IL2/JES6-1 timecourse]

Interleukin-2 (IL2) is the key trophic factor for T Regulatory (Treg) cells, regulating both their differentiation and homeostasis. To understand how temporally regulated responses to IL2 unfold in Tregs, we performed fine time-course analyses at single-cell level induced by IL2 in Tregs in vivo. The data revealed responses that were largely uniform in resting Treg, but diverse among activated or effector Tregs, matching different STAT5 signal transduction efficiency. Individual Tregs displayed divergences in the preponderance of changes downstream of STAT1 or STAT5. Overall design: We investigated how temporal interleukin-2 stimulation affect regulatory T cell gene expression. To do so, we stimulated FOXP3-GFP mice in vivo with IL2-JES6-1 complex at different time points between 1 to 128hrs and isolated splenic Tregs for each timepoints with PBS control. Cells were encapsulated using the Chromium Single Cell 5' v3 platform (10X Genomics). TotalSeq-A hashtag antibodies and samples (0.5 µl per sample for 20 min, BioLegend; hashtag 2, 128hr IL2-treated; hashtag 4, 128hr PBS-treated; hashtag 5, 64hr IL2-treated; hashtag 6, 24hr IL2-treated; hashtag 8, 16hr IL2-treated; hashtag 9, 8hrs IL2-treated; hashtag 10, 4hr IL2-treated; hashtag 11, 2hr IL2-treated; hashtag 12, 2h PBS-treated; hashtag 13, 1hr IL2-treated.