title: Saposins modulate human invariant Natural Killer T cells self-reactivity and facilitate lipid exchange with CD1d molecules during antigen presentation.
abstract: Lipid transfer proteins, such as molecules of the saposin family, facilitate extraction of lipids from biological membranes for their loading onto CD1d molecules. Although it has been shown that prosaposin-deficient mice fail to positively select invariant natural killer T (iNKT) cells, it remains unclear whether saposins can facilitate loading of endogenous iNKT cell agonists in the periphery during inflammatory responses. In addition, it is unclear whether saposins, in addition to loading, also promote dissociation of lipids bound to CD1d molecules. To address these questions, we used a combination of cellular assays and demonstrated that saposins influence CD1d-restricted presentation to human iNKT cells not only of exogenous lipids but also of endogenous ligands, such as the self-glycosphingolipid β-glucopyranosylceramide, up-regulated by antigen-presenting cells following bacterial infection. Furthermore, we demonstrated that in human myeloid cells CD1d-loading of endogenous lipids after bacterial infection, but not at steady state, requires trafficking of CD1d molecules through an endo-lysosomal compartment. Finally, using BIAcore assays we demonstrated that lipid-loaded saposin B increases the off-rate of lipids bound to CD1d molecules, providing important insights into the mechanisms by which it acts as a "lipid editor," capable of fine-tuning loading and unloading of CD1d molecules. These results have important implications in understanding how to optimize lipid-loading onto antigen-presenting cells, to better harness iNKT cells central role at the interface between innate and adaptive immunity.
authors.affiliation: Medical Research Council Human Immunology Unit, Radcliffe Department of Medicine, Medical Research Council Weatherall Institute of Molecular Medicine, University of Oxford, Headington, Oxford OX3 9DS, United Kingdom.
authors.forename: Mariolina
authors.initials: M
authors.lastname: Salio
authors.forename: Hemza
authors.initials: H
authors.lastname: Ghadbane
authors.forename: Omer
authors.initials: O
authors.lastname: Dushek
authors.forename: Dawn
authors.initials: D
authors.lastname: Shepherd
authors.forename: Jeremy
authors.initials: J
authors.lastname: Cypen
authors.forename: Uzi
authors.initials: U
authors.lastname: Gileadi
authors.forename: Michael C
authors.initials: MC
authors.lastname: Aichinger
authors.forename: Giorgio
authors.initials: G
authors.lastname: Napolitani
authors.forename: Xiaoyang
authors.initials: X
authors.lastname: Qi
authors.forename: P Anton
authors.initials: PA
authors.lastname: van der Merwe
authors.forename: Justyna
authors.initials: J
authors.lastname: Wojno
authors.forename: Natacha
authors.initials: N
authors.lastname: Veerapen
authors.forename: Liam R
authors.initials: LR
authors.lastname: Cox
authors.forename: Gurdyal S
authors.initials: GS
authors.lastname: Besra
authors.forename: Weiming
authors.initials: W
authors.lastname: Yuan
authors.forename: Peter
authors.initials: P
authors.lastname: Cresswell
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Cerundolo
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology; D055610:Antigens, CD1d / Q000378:metabolism*; D001419:Bacteria / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D057809:HEK293 Cells; D006801:Humans; D007113:Immunity, Innate / Q000276:immunology*; D047468:Immunoprecipitation; D050356:Lipid Metabolism / Q000502:physiology*; D055611:Natural Killer T-Cells / Q000276:immunology*; D049231:Saposins / Q000378:metabolism*; D012588:Scintillation Counting
chemical_list: D055610:Antigens, CD1d; C526877:CD1D protein, human; D049231:Saposins
keywords: autoreactivity; inflammation; innate immunity; lipid binding proteins