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

NF-κB, p38 MAPK, ERK and JNK activation have been implicated in IL-1R1-mediated immune cell activation [15] [16] [17] [18] [19] [20] [21] [22] [23] . Notably, the majority of these are murine studies. Based on these findings, we developed a customized 35 antibody IL-1β-response CyTOF panel to generate the first comprehensive atlas of IL-1β-induced signaling in peripheral blood mononuclear cells (PBMCs) in humans. Results demonstrated that IL-1β rapidly induces p-NF-kB and p-p38 in distinct effector memory (EM) T subsets, monocytes, dendritic cells (DC), natural killer (NK) cells and lineage-(Lin -)CD161 + CD25 + cells. IL-1βinduced increase in p-p38 was much less robust than p-NF-kB. In contrast to the rapid increase of p-NF-kB with only 15 minutes of IL-1β stimulation, prolonged stimulation for one hour greatly increased p-STAT3 and to a lesser extent p-STAT1 and p-STAT5 in subsets of T cells, monocytes, DCs and Lin -CD161 + CD25 + cells. Notably, we identified CCR6 as a marker to distinctly identify T cells most responsive to IL-1β. The IL-1R antagonist Anakinra significantly inhibited IL-1β-induced p-NF-kB in the CCR6 + T cells and CD11c myeloid DCs (mDC) with only a trending inhibition in CD14 monocytes and Lin -CD161 + CD25 + cells. This knowledge of the cell types, unique markers enriched on responsive cells and temporal expression of inflammatory signaling molecules in response to IL-1β stimulation in humans provides unique information that may be used to discover new approaches to limit IL-1β-Data obtained from the Helios instrument were in the form of .fcs files format. Samples were normalized using the Nolan lab MATLAB normalizer (http://github.com/nolanlab/bead-normalization/releases) and debarcoded using Zunder's lab debarcoder 24 (https://github.com/zunderlab/single-cell-debarcoder).