Purinergic P2X4 receptors and mitochondrial ATP production regulate T cell migration
Carola Ledderose, … , Gary A. Visner, Wolfgang G. Junger
Carola Ledderose, … , Gary A. Visner, Wolfgang G. Junger
Published June 12, 2018
Citation Information: J Clin Invest. 2018;128(8):3583-3594. https://doi.org/10.1172/JCI120972.
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Research Article Cell biology

Purinergic P2X4 receptors and mitochondrial ATP production regulate T cell migration

Abstract

T cells must migrate in order to encounter antigen-presenting cells (APCs) and to execute their varied functions in immune defense and inflammation. ATP release and autocrine signaling through purinergic receptors contribute to T cell activation at the immune synapse that T cells form with APCs. Here, we show that T cells also require ATP release and purinergic signaling for their migration to APCs. We found that the chemokine stromal-derived factor-1α (SDF-1α) triggered mitochondrial ATP production, rapid bursts of ATP release, and increased migration of primary human CD4+ T cells. This process depended on pannexin-1 ATP release channels and autocrine stimulation of P2X4 receptors. SDF-1α stimulation caused localized accumulation of mitochondria with P2X4 receptors near the front of cells, resulting in a feed-forward signaling mechanism that promotes cellular Ca2+ influx and sustains mitochondrial ATP synthesis at levels needed for pseudopod protrusion, T cell polarization, and cell migration. Inhibition of P2X4 receptors blocked the activation and migration of T cells in vitro. In a mouse lung transplant model, P2X4 receptor antagonist treatment prevented the recruitment of T cells into allograft tissue and the rejection of lung transplants. Our findings suggest that P2X4 receptors are therapeutic targets for immunomodulation in transplantation and inflammatory diseases.

Authors

Carola Ledderose, Kaifeng Liu, Yutaka Kondo, Christian J. Slubowski, Thomas Dertnig, Sara Denicoló, Mona Arbab, Johannes Hubner, Kirstin Konrad, Mahtab Fakhari, James A. Lederer, Simon C. Robson, Gary A. Visner, Wolfgang G. Junger

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Figure 1

ATP release through pannexin-1 channels is required for T cell migration.

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ATP release through pannexin-1 channels is required for T cell migration...
(A) ATP release at the cell surface from human CD4+ T cells before and 1 minute after stimulation with SDF-1α was visualized with the ATP probe 2-2Zn (left column; ×100 objective; scale bar, 10 μm). Cell migration was tracked for 30 minutes in the presence or absence of SDF-1α. Paths of individual cells superimposed (center column; ×63 objective; scale bar, 10 μm) or aligned with their origins at x = y = 0 (right column) are shown. Data are representative of 5 experiments. (See also Supplemental Video 1). (B) Migration speed and ATP release of CD4+ T cells in response to SDF-1α. (C) Speed and migration range of CD4+ T cells treated with increasing concentrations of SDF-1α (30-minute observation). (D) CD4+ T cells were stained with 2-2Zn and the response to CBX (100 μM) or SDF-1α was analyzed with fluorescence microscopy. Representative images before, 0.5 minutes after addition of CBX, and 1 minute after addition of SDF-1α (left) and traces of cells derived from 2 separate experiments (control, n = 31; CBX, n = 39) are shown. Data are mean ± SD; ×100 objective; scale bar, 10 μm. (See also Supplemental Video 2.) (E) ATP concentrations in the supernatants of CD4+ T cells treated with CBX (50 μM) or 10panx1 (100 μM) and stimulated with SDF-1α for 5 minutes. (F) Spontaneous or SDF-1α–induced migration speed of Jurkat cells after silencing of PANX1 or treatment with CBX (100 μM; see also Supplemental Video 3; *P < 0.05, #P < 0.05 by Kruskal-Wallis test). Data in B (migration speed), C, and F represent mean ± SD of 60 cells analyzed in 3 independent experiments. Data in B (ATP release) and E represent mean ± SD of 3 independent experiments; *P < 0.05 vs. control (1-way ANOVA); #P < 0.05 (unpaired 2-tailed t test).