title: An essential role of syntaxin 3 protein for granule exocytosis and secretion of IL-1α, IL-1β, IL-12b, and CCL4 from differentiated HL-60 cells.
abstract: Besides their roles in the killing of pathogens, neutrophils have the capacity to package a variety of cytokines into cytoplasmic granules for subsequent release upon inflammatory conditions. Because the rapid secretion of cytokines orchestrates the action of other immune cells at the infection site and thus, can contribute to the development and chronicity of inflammatory diseases, we aimed to determine the intracellular SNARE machinery responsible for the regulation of cytokine secretion and degranulation. From a constructed gene-expression network, we first selected relevant cytokines for functional validation by the CBA approach. We established a cytokine-secretion profile for human neutrophils and dHL-60 cells, underlining their similar ability to secrete a broad variety of cytokines within proinflammatory conditions mimicked by LPS stimulation. Secondly, after screening of SNARE genes by microarray experiments, we selected STX3 for further functional studies. With the use of a siRNA strategy, we show that STX3 is clearly required for the maximal release of IL-1α, IL-1β, IL-12b, and CCL4 without alteration of other cytokine secretion in dHL-60 cells. In addition, we demonstrate that STX3 is involved in MMP-9 exocytosis from gelatinase granules, where STX3 is partly localized. Our results suggest that the secretion of IL-1α, IL-1β, IL-12b, and CCL4 occurs during gelatinase degranulation, a process controlled by STX3. In summary, these findings provide first evidence that STX3 has an essential role in trafficking pathways of cytokines in neutrophil granulocytes.
authors.affiliation: *University of Luxembourg, Life Sciences Research Unit, Calcium Signaling and Inflammation, Luxembourg; and Centre de Recherche Public Santé, Luxembourg.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Naegelen
authors.affiliation: *University of Luxembourg, Life Sciences Research Unit, Calcium Signaling and Inflammation, Luxembourg; and Centre de Recherche Public Santé, Luxembourg.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Plançon
authors.affiliation: *University of Luxembourg, Life Sciences Research Unit, Calcium Signaling and Inflammation, Luxembourg; and Centre de Recherche Public Santé, Luxembourg.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Nicot
authors.affiliation: *University of Luxembourg, Life Sciences Research Unit, Calcium Signaling and Inflammation, Luxembourg; and Centre de Recherche Public Santé, Luxembourg.
authors.forename: Tony
authors.initials: T
authors.lastname: Kaoma
authors.affiliation: *University of Luxembourg, Life Sciences Research Unit, Calcium Signaling and Inflammation, Luxembourg; and Centre de Recherche Public Santé, Luxembourg.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Muller
authors.affiliation: *University of Luxembourg, Life Sciences Research Unit, Calcium Signaling and Inflammation, Luxembourg; and Centre de Recherche Public Santé, Luxembourg.
authors.forename: Laurent
authors.initials: L
authors.lastname: Vallar
authors.affiliation: *University of Luxembourg, Life Sciences Research Unit, Calcium Signaling and Inflammation, Luxembourg; and Centre de Recherche Public Santé, Luxembourg.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Tschirhart
authors.affiliation: *University of Luxembourg, Life Sciences Research Unit, Calcium Signaling and Inflammation, Luxembourg; and Centre de Recherche Public Santé, Luxembourg sabrina.brechard@uni.lu.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Bréchard
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects; D054407:Chemokine CCL4 / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects; D005089:Exocytosis / Q000187:drug effects; D018093:Gelatinases / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D055785:Gene Knockdown Techniques; D015894:Genome, Human; D018922:HL-60 Cells; D006801:Humans; D007249:Inflammation / Q000473:pathology; D053762:Interleukin-12 Subunit p40 / Q000378:metabolism*; D053582:Interleukin-1alpha / Q000378:metabolism*; D053583:Interleukin-1beta / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism; D050765:Qa-SNARE Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: C066507:CCL4 protein, human; D054407:Chemokine CCL4; C500879:IL12B protein, human; D053762:Interleukin-12 Subunit p40; D053582:Interleukin-1alpha; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D050765:Qa-SNARE Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D018093:Gelatinases
keywords: SNARE proteins; cytokines; extracellular release; gelatinase granules; neutrophils