title: Conserved and heterogeneous lipid antigen specificities of CD1d-restricted NKT cell receptors.
abstract: CD1d-restricted NKT cells use structurally conserved TCRs and recognize both self and foreign glycolipids, but the TCR features that determine these Ag specificities remain unclear. We investigated the TCR structures and lipid Ag recognition properties of five novel Valpha24-negative and 13 canonical Valpha24-positive/Vbeta11-positive human NKT cell clones generated using alpha-galactosylceramide (alpha-GalCer)-loaded CD1d tetramers. The Valpha24-negative clones expressed Vbeta11 paired with Valpha10, Valpha2, or Valpha3. Strikingly, their Valpha-chains had highly conserved rearrangements to Jalpha18, resulting in CDR3alpha loop sequences that are nearly identical to those of canonical TCRs. Valpha24-positive and Valpha24-negative clones responded similarly to alpha-GalCer and a closely related bacterial analog, suggesting that conservation of the CDR3alpha loop is sufficient for recognition of alpha-GalCer despite CDR1alpha and CDR2alpha sequence variation. Unlike Valpha24-positive clones, the Valpha24-negative clones responded poorly to a glucose-linked glycolipid (alpha-glucosylceramide), which correlated with their lack of a conserved CDR1alpha amino acid motif, suggesting that fine specificity for alpha-linked glycosphingolipids is influenced by Valpha-encoded TCR regions. Valpha24-negative clones showed no response to isoglobotrihexosylceramide, indicating that recognition of this mammalian lipid is not required for selection of Jalpha18-positive TCRs that can recognize alpha-GalCer. One alpha-GalCer-reactive, Valpha24-positive clone differed from the others in responding specifically to mammalian phospholipids, demonstrating that semi-invariant NKT TCRs have a capacity for private Ag specificities that are likely conferred by individual TCR beta-chain rearrangements. These results highlight the variation in Ag recognition among CD1d-restricted TCRs and suggest that TCR alpha-chain elements contribute to alpha-linked glycosphingolipid specificity, whereas TCR beta-chains can confer heterogeneous additional reactivities.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin Medical School, 1300 University Avenue, Madison, WI 53706, USA.
authors.forename: Manfred
authors.initials: M
authors.lastname: Brigl
authors.forename: Peter
authors.initials: P
authors.lastname: van den Elzen
authors.forename: Xiuxu
authors.initials: X
authors.lastname: Chen
authors.forename: Jennifer Hartt
authors.initials: JH
authors.lastname: Meyers
authors.forename: Douglass
authors.initials: D
authors.lastname: Wu
authors.forename: Chi-Huey
authors.initials: CH
authors.lastname: Wong
authors.forename: Faye
authors.initials: F
authors.lastname: Reddington
authors.forename: Petr A
authors.initials: PA
authors.lastname: Illarianov
authors.forename: Gurdyal S
authors.initials: GS
authors.lastname: Besra
authors.forename: Michael B
authors.initials: MB
authors.lastname: Brenner
authors.forename: Jenny E
authors.initials: JE
authors.lastname: Gumperz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018949:Antigens, CD1 / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D055610:Antigens, CD1d; D001324:Autoantigens / Q000276:immunology; D002478:Cells, Cultured; D018360:Crystallography, X-Ray; D016207:Cytokines / Q000378:metabolism; D006801:Humans; D008055:Lipids / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry / Q000276:immunology; D011971:Receptors, Immunologic / Q000276:immunology* / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism*
chemical_list: D018949:Antigens, CD1; D055610:Antigens, CD1d; D001324:Autoantigens; C526877:CD1D protein, human; D016207:Cytokines; D008055:Lipids; D011948:Receptors, Antigen, T-Cell; D011971:Receptors, Immunologic