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

Kothari, H.; Williams, C. M.; McSkimming, C.; Vigneshwar, M.; Zunder, E. R.; McNamara, C. A.

doi:10.1101/2020.04.19.047274

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bioRxiv · 10.1101/2020.04.19.047274

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

Kothari, H.·Williams, C. M.·McSkimming, C.·Vigneshwar, M.·Zunder, E. R.·McNamara, C. A.

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Abstract

IL-1{beta} has emerged as a key mediator of the cytokine storm linked to high morbidity and mortality from COVID-19 and blockade of the IL-1 receptor (IL-1R) with Anakinra has entered clinical trials in COVID-19 subjects. Yet, knowledge of the specific immune cell subsets targeted by IL-1{beta} and IL-1{beta}-induced signaling pathways in humans is limited. Utilizing mass cytometry (CyTOF) of human peripheral blood mononuclear cells, we identified effector memory CD4 T cells and CD4-CD8low/-CD161+ T cells as the circulating immune subtypes with the greatest expression of p-NF-{kappa}B in response to IL-1{beta} stimulation. Notably, CCR6 distinctly identified T cells most responsive to IL-1{beta}. Other subsets including CD11c myeloid dendritic cells (mDCs), classical monocytes (CM), two subsets of natural killer cells (CD16-CD56brightCD161- and CD16-CD56dimCD161+) and a population of lineage-(Lin-) cells expressing CD161 and CD25 also showed IL-1{beta}-induced expression of p-NF-kB. The IL-1R antagonist, Anakinra significantly inhibited IL-1{beta}-induced p-NF-kB in the CCR6+ T cells and CD11c mDCs with a trending inhibition in CD14 monocytes and Lin-CD161+CD25+ cells. IL-1{beta} also induced a rapid but much less robust increase in p-p38 expression as compared to p-NF-kB in the majority of these same immune cell subsets. Prolonged IL-1{beta} stimulation greatly increased p-STAT3 and to a much lesser extent p-STAT1 and p-STAT5 in T cell subsets, monocytes, DCs and the Lin-CD161+CD25+ cells suggesting IL-1{beta}-induced production of downstream STAT-activating cytokines, consistent with its role in cytokine storm. Interindividual heterogeneity and inhibition of this activation by Anakinra raises the intriguing possibility that assays to measure IL-1{beta}-induced p-NF-kB in CCR6+ T cell subtypes could identify those at higher risk of cytokine storm and those most likely to benefit from Anakinra therapy.

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Kothari, H., Williams, C. M., McSkimming, C., Vigneshwar, M., Zunder, E. R., McNamara, C. A.. 2020-04-19. Identification of Human Immune Cell Subtypes Most Vulnerable to IL-1β-induced Inflammatory Signaling Using Mass Cytometry. https://doi.org/10.1101/2020.04.19.047274

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