Signaling and functional competency of neutrophils derived from bone-marrow cells expressing the ER-HOXB8 oncoprotein

Article Article scientifique Article Signaling and functional competency of neutrophils derived from bone-marrow cells expressing the ER-HOXB8 oncoprotein eng 2019-06-19 2019-11 author Saul Stéphanie 839895 author Castelbou Cyril 30263 author Fickentscher Céline author Demaurex Nicolas 0000-0002-9933-6772 24642 1101-1115 10.1002/JLB.2A0818-314R 31216372 0741-5400 Neutrophils play a central role in immunity and inflammation via their intrinsic ability to migrate into inflamed tissue, to phagocytose pathogens, and to kill bacterial and fungi by releasing large quantities of superoxide anions and lytic enzymes. The molecular pathways controlling neutrophil microbicidal functions are still unclear, because neutrophils have a short half-life and are resistant to genetic manipulation. Neutrophil-like cells (NLC) can be generated from myeloid progenitors conditionally immortalized with the ER-HoxB8 oncoprotein, but whether these cells can replace neutrophils in high-throughput functional assays is unclear. Here, we assess the ability of NLC derived from ER-HoxB8 progenitors to produce ROS and to perform chemotaxis and phagocytosis. We compare the Ca2+responses and effector functions of NLC to primary murine neutrophils and document the molecular basis of their functional differences by mRNA profiling. Pro-inflammatory cytokines enhanced the expression by NLC of neutrophil surface markers and transcription factors. Ca2+elevations evoked in NLC by agonists, adhesion receptors, and store depletion resembled the physiological responses recorded in primary neutrophils, but NLC expressed reduced amounts of Ca2+signaling proteins and of chemotactic receptors. Unlike their myeloid progenitors, NLC produced H2O2when adhered to fibronectin, migrated toward chemotactic peptides, phagocytosed opsonized particles, and generated intracellular ROS. NLC phagocytosed as efficiently as primary neutrophils but produced 50 times less ROS and migrated less efficiently toward chemoattractant. Our data indicate that NLC can replace neutrophils to study Ca2+signaling and phagocytosis, but that their incomplete granulocytic differentiation limits their use for chemotaxis and ROS production assays. Granulocytes In vitro differentiation Myeloid cell lines Polymorphonuclear cells Animals Bone Marrow Cells / cytology Bone Marrow Cells / immunology Calcium Signaling / genetics Calcium Signaling / immunology Gene Expression Regulation / immunology Homeodomain Proteins / genetics Homeodomain Proteins / immunology Male Mice Mice, Transgenic Neutrophils / cytology Neutrophils / immunology Oncogene Proteins / genetics Oncogene Proteins / immunology Reactive Oxygen Species / immunology Signal Transduction / genetics Signal Transduction / immunology Dewey 612 Swiss National Science Foundation Calcium signaling at membrane contact sites during phagocytosis 169491 CH Projects Swiss National Science Foundation Store-operated calcium channels in health and disease 160782 CH Programmes 134 Département de physiologie cellulaire et métabolisme 21C false 168 Signaux intracellulaires 210 Journal of leukocyte biology 106 5 dc119a52-c94a-4611-8bcc-4648336ac9d7 Article (Published version) Public Article-scientifique-from-unpaywall.pdf 2511877 application/pdf CC BY openAccess Faculté de médecine / Section de médecine fondamentale / Département de physiologie cellulaire et métabolisme

Title	Published in	Year	Views	Downloads

Signaling and functional competency of neutrophils derived from bone-marrow cells expressing the ER-HOXB8 oncoprotein	Journal of leukocyte biology	2019	136	211
The role of STIM proteins in neutrophil functions	The Journal of Physiology	2018	370	282

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Saul, Stéphanie