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 3

T cell migration and activation depends on P2X4 receptors.

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T cell migration and activation depends on P2X4 receptors. (A) CD69 expr...
(A) CD69 expression in CD4+ T cells stimulated for 5 hours with SDF-1α and anti-CD3 antibodies in a PBMC culture was measured by flow cytometry. Positive controls (stimulation with anti-CD3/anti-CD28 coated beads) and negative controls (no stimulation or stimulation with anti-CD3 in monocyte-depleted cultures) were included as indicated. Data represent mean ± SD of 3 individual experiments. *P < 0.05 vs. 0 ng/ml SDF-1α (1-way ANOVA); TCR, T cell receptor. (B) PBMCs were placed into fibronectin-coated glass-bottom chamber slides, stained with APC-labeled anti-CD4 antibodies, stimulated with SDF-1α, and migration speed of CD4+ T cells was analyzed by time-lapse microscopy. Data are mean ± SD of 50 cells analyzed in 3 separate experiments. CD69 expression following stimulation with SDF-1α and anti-CD3 antibodies for 5 hours was analyzed as in A. Data represent mean ± SD of 3 separate experiments. (C) Correlation between CD69 expression and migration speed. Data are the mean values of the experiments shown in B. (D) CD4+ T cells were treated with CCCP (5 μM), CBX (100 μM), suramin (100 μM), or inhibitors of P2X1 (NF023; 10 μM), P2X4 (5-BDBD; 10 μM), or P2X7 (A438079; 10 μM) receptors, and migration speed in response to SDF-1α was analyzed. Data represent mean ± SD of 80 cells analyzed in 3 experiments; #P < 0.05 vs. control (Kruskal-Wallis test). (E) CD69 expression following TCR stimulation with anti-CD3 for 3 hours was analyzed as in A. (F) Proliferation of CD4+ T cells in a PBMC culture stimulated with anti-CD3 antibodies for 72 hours was determined by analyzing CFSE dilution. Data in E and F represent mean ± SD of 6 (E) or 3 (F) individual experiments. *P < 0.05 vs. control (1-way ANOVA).