Purinergic Signaling: A Fundamental Mechanism in Neutrophil Activation
Feedback for Function
Neutrophils migrate to sites of infection, where they kill pathogens by processes such as phagocytosis and the release of reactive oxygen species. However, activated neutrophils can also result in tissue damage and inflammatory diseases in the host; thus, a better understanding of the mechanisms that regulate neutrophil activation could help in the development of therapies that could curb their destructive side effects. Chen et al. found that neutrophils responded to a range of infectious and inflammatory signals by releasing adenosine triphosphate (ATP). In addition to its role as a cellular energy source, ATP and its metabolites function as intercellular signaling molecules by stimulating purinergic receptors. The authors found that stimulation of formyl peptide receptors (FPRs) on neutrophils triggered the release, through pannexin-1 hemichannels, of ATP that signaled in an autocrine fashion through P2Y2 receptors. Moreover, this autocrine signal was required for neutrophil activation. In addition, mice deficient in P2Y2 receptors were less capable of clearing bacteria than were their wild-type counterparts. Together, these data suggest that feedback signaling by ATP released by neutrophils contributes to their activation.
Abstract
Efficient activation of neutrophils is a key requirement for effective immune responses. We found that neutrophils released cellular adenosine triphosphate (ATP) in response to exogenous stimuli such as formylated bacterial peptides and inflammatory mediators that activated Fcγ, interleukin-8, C5a complement, and leukotriene B4 receptors. Stimulation of the formyl peptide receptor (FPR) led to ATP release through pannexin-1 (panx1) hemichannels, and FPRs colocalized with P2Y2 nucleotide receptors on the cell surface to form a purinergic signaling system that facilitated neutrophil activation. Disruption of this purinergic signaling system by inhibiting or silencing panx1 hemichannels or P2Y2 receptors blocked neutrophil activation and impaired innate host responses to bacterial infection. Thus, purinergic signaling is a fundamental mechanism required for neutrophil activation and immune defense.
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Supplementary Material
Summary
Fig. S1. Colocalization of FPR and TTYH3 in human PMNs.
Fig. S2. Effects of blockers of ATP release on the ability of PMNs to hydrolyze extracellular ATP.
Fig. S3. Effects of blockers of ATP release on ATP-induced Ca2+ mobilization.
Fig. S4. Knockdown of P2Y2 and TTYH3.
Fig. S5. Colocalization of P2Y2 and FPR.
Movie S1. Z-stack view of a dHL-60 cell cotransfected with plasmids encoding P2Y2-ECFP (green) and FPR-EYFP (red).
Movie S2. A 3D view of a dHL-60 cell cotransfected with plasmids encoding P2Y2-ECFP (green) and FPR-EYFP (red).
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Information & Authors
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Published In
Science Signaling
Volume 3 | Issue 125
June 2010
June 2010
Copyright
Copyright © 2010, American Association for the Advancement of Science.
Submission history
Received: 21 July 2009
Accepted: 20 May 2010
Acknowledgments
Acknowledgments: We thank H. Sitte and C. Nanoff (Medical University of Vienna, Vienna, Austria) for providing the pECFP-P2Y2 receptor construct. Funding: This study was supported in part by a Shock Society/Nova Nordisk research grant for Early Career Investigators (Y.C.) and by NIH grants GM-51477, GM-60475, AI-072287, and AI-080582 and Congressionally Directed Medical Research Programs grant PR043034 (W.G.J.). Author contributions: Y.C. performed most of the experiments and prepared the manuscript and figures; Y.Y. and U.K.T. provided technical and logistical assistance; Y.S., T.W., Y.I., and A.E. performed the animal experiments; A.L. and Q.Z. performed calcium measurements; C.H. helped with the discussion and interpretation of results; and W.G.J. was responsible for the design and coordination of the study and finalized the manuscript. Competing interests: The authors have no conflicts of interest to declare. Use of the expression plasmids described in this study requires a materials transfer agreement.
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