Identification of Human Immune Cell Subtypes Most Vulnerable to IL-1ß-induced Inflammatory Signaling Using Mass Cytometry

Hema Kothari; Corey M. Williams; Chantel McSkimming; Mythili Vigneshwar; Eli R. Zunder; Coleen A. McNamara

To identify specific T cell subsets that show IL-1β-induced increase in p-NF-kB and p-p38, we performed Uniform Manifold Approximation and Projection (UMAP), followed by Leiden clustering on CD3 + T cells (Supplementary Fig. 3d ). Unsupervised clustering of CD3 + T cells resulted in 12 clusters (Fig. 2a) . The frequency of each cluster is shown in Fig. 2b . Clusters were annotated based on the expression of lineage markers as shown in the heatmap in Fig. 2c . A marked increase in the frequency of p-NF-kB + cells was observed in TH17 cells (cluster 3; CD4 + CD45RO + CCR6 + CD161 + CD26 + ), T follicular helper cells (Tfh) (cluster 4; CD4 + CD45RO + CXCR5 + ), CD8 low/-CD161 + T cells (cluster 8) and memory T regulatory cells (Mtregs) (cluster 12; CD4 + CD45RO + CD25 + CD127 low/-) (Fig. 2d) . Cluster 8 showed a bimodal expression of CD8 and biaxial gating demonstrated presence of two distinct cell populations within cluster 8: CD8 -(57.6 ± 2.00%; SEM) and CD8 + (40.5 ± 2.16%; SEM) that were both CD4 -CD45RO + , expressed comparable levels of CD161, and showed IL-1β-induced expression of p-NF-kB (Supplementary Fig. 7a-c and i) . Moreover, similar to TH17 cells (cluster 3), both CD8and CD8 + subpopulations within cluster 8 expressed CD26 (Supplementary Fig. 7d) , a marker that is associated with cells capable of producing IL-17 29 . Notably, all four IL-1β responsive T cell clusters author/funder. All rights reserved. No reuse allowed without permission.