title: N-Formyl peptides drive mitochondrial damage associated molecular pattern induced neutrophil activation through ERK1/2 and P38 MAP kinase signalling pathways.
abstract: Traumatic injury results in a systemic inflammatory response syndrome (SIRS), a phenomenon characterised by the release of pro-inflammatory cytokines into the circulation and immune cell activation. Released from necrotic cells as a result of tissue damage, damage associated molecular patterns (DAMPs) are thought to initiate the SIRS response by activating circulating immune cells through surface expressed pathogen recognition receptors. Neutrophils, the most abundant leucocyte in human circulation, are heavily implicated in the initial immune response to traumatic injury and have been shown to elicit a robust functional response to DAMP stimulation. Here, we confirm that mitochondrial DAMPs (mtDAMPs) are potent activators of human neutrophils and show for the first time that signalling through the mitogen-activated-protein-kinases p38 and extracellular-signal-related-kinase 1/2 (ERK1/2) is essential for this response. At 40 and/or 100 μg/ml, mtDAMPs activated human neutrophils, indicated by a significant reduction in the surface expression of L-selectin, and triggered a number of functional responses from both resting and tumour necrosis factor-α primed neutrophils, which included reactive oxygen species (ROS) generation, degranulation, secretion of interleukin-8 and activation of p38 and ERK1/2 MAPKs. Pre-treatment of neutrophils with Cyclosporin H, a selective inhibitor of formyl peptide receptor-1 (FPR-1), significantly inhibited mtDAMP-induced L-selectin shedding as well as p38 and ERK1/2 activation, suggesting that N-formyl peptides are the main constituents driving mtDAMP-induced neutrophil activation. Indeed, no evidence of L-selectin shedding or p38 and ERK1/2 activation was observed in neutrophils challenged with mitochondrial DNA alone. Interestingly, pharmacological inhibition of p38 or ERK1/2 either alone or in combination significantly inhibited L-selectin shedding and IL-8 secretion by mtDAMP-challenged neutrophils, revealing for the first time that MAPK activation is required for mtDAMP-induced neutrophil activation and function. Our findings demonstrate that signalling through FPR-1 and activation of p38 and ERK1/2 MAPKs are key events in mtDAMP-induced neutrophil activation. Gaining an understanding of the signalling pathways involved in mtDAMP-induced neutrophil activation may assist in the development of future therapeutic strategies aimed at targeting the SIRS response to improve the outcome of the hospitalised trauma patient. Reducing the severity of the inflammatory response may realise substantial benefits for the severely injured trauma patient.
authors: NIHR Surgical Reconstruction and Microbiology Research Centre, School of Immunity and Infection, Birmingham University Medical School, Birmingham B15 2TT, UK. Electronic address: J.Hazeldine@bham.ac.uk.
authors: Jon
authors: J
authors: Hazeldine
authors: NIHR Surgical Reconstruction and Microbiology Research Centre, School of Immunity and Infection, Birmingham University Medical School, Birmingham B15 2TT, UK; Healing Foundation Centre for Burns Research, Queen Elizabeth Hospital, Birmingham B15 2WB, UK.
authors: Peter
authors: P
authors: Hampson
authors: Royal Centre for Defence Medicine, Birmingham Research Park, Birmingham B15 2SQ, UK.
authors: Francis Adusei
authors: FA
authors: Opoku
authors: Royal Centre for Defence Medicine, Birmingham Research Park, Birmingham B15 2SQ, UK.
authors: Mark
authors: M
authors: Foster
authors: NIHR Surgical Reconstruction and Microbiology Research Centre, School of Immunity and Infection, Birmingham University Medical School, Birmingham B15 2TT, UK.
authors: Janet M
authors: JM
authors: Lord
mesh_terms: D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D019041:L-Selectin / Q000378:metabolism; D020935:MAP Kinase Signaling System; D008928:Mitochondria / Q000378:metabolism* / Q000473:pathology; D024101:Mitochondrial Proteins / Q000378:metabolism*; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology; D010766:Phosphorylation; D015398:Signal Transduction; D018746:Systemic Inflammatory Response Syndrome / Q000209:etiology / Q000276:immunology* / Q000628:therapy; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014947:Wounds and Injuries / Q000150:complications / Q000276:immunology* / Q000628:therapy; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: C508606:CXCL8 protein, human; D016209:Interleukin-8; D024101:Mitochondrial Proteins; C439524:TNF protein, human; D014409:Tumor Necrosis Factor-alpha; D019041:L-Selectin; D048051:p38 Mitogen-Activated Protein Kinases
keywords: Mitochondrial damage associated molecular patterns; Mitogen activated protein kinases; Neutrophils; Trauma