title: Genome-wide siRNA screen reveals a new cellular partner of NK cell receptor KIR2DL4: heparan sulfate directly modulates KIR2DL4-mediated responses.
abstract: KIR2DL4 (CD158d) is a distinct member of the killer cell Ig-like receptor (KIR) family in human NK cells that can induce cytokine production and cytolytic activity in resting NK cells. Soluble HLA-G, normally expressed only by fetal-derived trophoblast cells, was reported to be a ligand for KIR2DL4; however, KIR2DL4 expression is not restricted to the placenta and can be found in CD56(high) subset of peripheral blood NK cells. We demonstrated that KIR2DL4 can interact with alternative ligand(s), expressed by cells of epithelial or fibroblast origin. A genome-wide high-throughput siRNA screen revealed that KIR2DL4 recognition of cell-surface ligand(s) is directly regulated by heparan sulfate (HS) glucosamine 3-O-sulfotransferase 3B1 (HS3ST3B1). KIR2DL4 was found to directly interact with HS/heparin, and the D0 domain of KIR2DL4 was essential for this interaction. Accordingly, exogenous HS/heparin can regulate cytokine production by KIR2DL4-expressing NK cells and HEK293T cells (HEK293T-2DL4), and induces differential localization of KIR2DL4 to rab5(+) and rab7(+) endosomes, thus leading to downregulation of cytokine production and degradation of the receptor. Furthermore, we showed that intimate interaction of syndecan-4 (SDC4) HS proteoglycan (HSPG) and KIR2DL4 directly affects receptor endocytosis and membrane trafficking.
authors.affiliation: The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
authors.forename: Michael
authors.initials: M
authors.lastname: Brusilovsky
authors.affiliation: The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
authors.forename: Moti
authors.initials: M
authors.lastname: Cordoba
authors.affiliation: The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
authors.forename: Benyamin
authors.initials: B
authors.lastname: Rosental
authors.affiliation: The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
authors.forename: Oren
authors.initials: O
authors.lastname: Hershkovitz
authors.affiliation: The Research Institute of Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Andrake
authors.affiliation: The Research Institute of Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Anna
authors.initials: A
authors.lastname: Pecherskaya
authors.affiliation: The Research Institute of Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Margret B
authors.initials: MB
authors.lastname: Einarson
authors.affiliation: The Research Institute of Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
authors.forename: Alex
authors.initials: A
authors.lastname: Braiman
authors.affiliation: The Research Institute of Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Kerry S
authors.initials: KS
authors.lastname: Campbell
authors.affiliation: The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, Beer-Sheva, Israel.|National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
authors.forename: Angel
authors.initials: A
authors.lastname: Porgador
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016466:CHO Cells; D002460:Cell Line; D003412:Cricetulus; D004705:Endocytosis; D057809:HEK293 Cells; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D034741:RNA, Small Interfering; D054345:Receptors, KIR2DL4 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015398:Signal Transduction / Q000276:immunology; D015238:Sulfotransferases / Q000378:metabolism*; D053671:Syndecan-4 / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D020696:rab5 GTP-Binding Proteins / Q000378:metabolism; D000090802:rab7 GTP-Binding Proteins
chemical_list: D000911:Antibodies, Monoclonal; D006493:Heparin; D006497:Heparan Sulfate; D034741:RNA, Small Interfering; D054345:Receptors, KIR2DL4; D015238:Sulfotransferases; D053671:Syndecan-4; D020691:rab GTP-Binding Proteins; D020696:rab5 GTP-Binding Proteins; D000090802:rab7 GTP-Binding Proteins; C515079:KIR2DL4 protein, human; C508304:SDC4 protein, human; C000716463:rab7 GTP-binding proteins, human; C021350:heparitin sulfotransferase