title: RNA-seq reveals activation of both common and cytokine-specific pathways following neutrophil priming.
abstract: Neutrophils are central to the pathology of inflammatory diseases, where they can damage host tissue through release of reactive oxygen metabolites and proteases, and drive inflammation via secretion of cytokines and chemokines. Many cytokines, such as those generated during inflammation, can induce a similar "primed" phenotype in neutrophils, but it is unknown if different cytokines utilise common or cytokine-specific pathways to induce these functional changes. Here, we describe the transcriptomic changes induced in control human neutrophils during priming in vitro with pro-inflammatory cytokines (TNF-α and GM-CSF) using RNA-seq. Priming led to the rapid expression of a common set of transcripts for cytokines, chemokines and cell surface receptors (CXCL1, CXCL2, IL1A, IL1B, IL1RA, ICAM1). However, 580 genes were differentially regulated by TNF-α and GM-CSF treatment, and of these 58 were directly implicated in the control of apoptosis. While these two cytokines both delayed apoptosis, they induced changes in expression of different pro- and anti-apoptotic genes. Bioinformatics analysis predicted that these genes were regulated via differential activation of transcription factors by TNF-α and GM-CSF and these predictions were confirmed using functional assays: inhibition of NF-κB signalling abrogated the protective effect of TNF-α (but not that of GM-CSF) on neutrophil apoptosis, whereas inhibition of JAK/STAT signalling abrogated the anti-apoptotic effect of GM-CSF, but not that of TNF-α (p<0.05). These data provide the first characterisation of the human neutrophil transcriptome following GM-CSF and TNF-α priming, and demonstrate the utility of this approach to define functional changes in neutrophils following cytokine exposure. This may provide an important, new approach to define the molecular properties of neutrophils after in vivo activation during inflammation.
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Liverpool, Merseyside, United Kingdom.
authors.forename: Helen L
authors.initials: HL
authors.lastname: Wright
authors.forename: Huw B
authors.initials: HB
authors.lastname: Thomas
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Moots
authors.forename: Steven W
authors.initials: SW
authors.lastname: Edwards
mesh_terms: D016207:Cytokines / Q000096:biosynthesis* / Q000276:immunology / Q000494:pharmacology; D005260:Female; D006801:Humans; D007249:Inflammation / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis* / Q000276:immunology; D053612:Janus Kinases / Q000276:immunology / Q000378:metabolism; D008297:Male; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D050791:STAT Transcription Factors / Q000276:immunology / Q000378:metabolism; D015398:Signal Transduction; D059467:Transcriptome / Q000187:drug effects / Q000276:immunology
chemical_list: D016207:Cytokines; C573420:ICAM1 protein, human; D050791:STAT Transcription Factors; D018799:Intercellular Adhesion Molecule-1; D053612:Janus Kinases
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