title: Activation of human γδ T cells by cytosolic interactions of BTN3A1 with soluble phosphoantigens and the cytoskeletal adaptor periplakin.
abstract: The three butyrophilin BTN3A molecules, BTN3A1, BTN3A2, and BTN3A3, are members of the B7/butyrophilin-like group of Ig superfamily receptors, which modulate the function of T cells. BTN3A1 controls activation of human Vγ9/Vδ2 T cells by direct or indirect presentation of self and nonself phosphoantigens (pAg). We show that the microbial metabolite (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate binds to the intracellular B30.2 domain of BTN3A1 with an affinity of 1.1 μM, whereas the endogenous pAg isopentenyl pyrophosphate binds with an affinity of 627 μM. Coculture experiments using knockdown cell lines showed that in addition to BTN3A1, BTN3A2 and BTN3A3 transmit activation signals to human γδ T cells in response to (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate and the aminobisphosphonate drug zoledronate that causes intracellular accumulation of isopentenyl pyrophosphate. The plakin family member periplakin, identified in yeast two-hybrid assays, interacted with a membrane-proximal di-leucine motif, located proximal to the B30.2 domain in the BTN3A1 cytoplasmic tail. Periplakin did not interact with BTN3A2 or BTN3A3, which do not contain the di-leucine motif. Re-expression into a BTN3A1 knockdown line of wild-type BTN3A1, but not of a variant lacking the periplakin binding motif, BTN3A1Δexon5, restored γδ T cell responses, demonstrating a functional role for periplakin interaction. These data, together with the widespread expression in epithelial cells, tumor tissues, and macrophages detected using BTN3A antiserum, are consistent with complex functions for BTN3A molecules in tissue immune surveillance and infection, linking the cell cytoskeleton to γδ T cell activation by indirectly presenting pAg to the Vγ9/Vδ2 TCR.
authors: Immunology Division, Department of Pathology, University of Cambridge, Cambridge Institute for Medical Research, Cambridge CB2 0XY, United Kingdom; dar32@cam.ac.uk.
authors: David A
authors: DA
authors: Rhodes
authors: Cardiff Institute of Infection & Immunity, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom; and.
authors: Hung-Chang
authors: HC
authors: Chen
authors: Protein and Nucleic Acid Chemistry Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors: Amanda J
authors: AJ
authors: Price
authors: Protein and Nucleic Acid Chemistry Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors: Anthony H
authors: AH
authors: Keeble
authors: Cardiff Institute of Infection & Immunity, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom; and.
authors: Martin S
authors: MS
authors: Davey
authors: Protein and Nucleic Acid Chemistry Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, United Kingdom.
authors: Leo C
authors: LC
authors: James
authors: Cardiff Institute of Infection & Immunity, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom; and.
authors: Matthias
authors: M
authors: Eberl
authors: Immunology Division, Department of Pathology, University of Cambridge, Cambridge Institute for Medical Research, Cambridge CB2 0XY, United Kingdom;
authors: John
authors: J
authors: Trowsdale
mesh_terms: D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D015703:Antigens, CD / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D001665:Binding Sites; D000071797:Butyrophilins; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D011756:Diphosphates / Q000494:pharmacology; D004164:Diphosphonates / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000276:immunology; D045782:Hemiterpenes / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D008958:Models, Molecular; D009943:Organophosphorus Compounds / Q000494:pharmacology; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D051179:Plakins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D061251:Primary Cell Culture; D011485:Protein Binding; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D020798:Two-Hybrid System Techniques; D000077211:Zoledronic Acid
chemical_list: C443995:4-hydroxy-3-methylbut-2-enyl pyrophosphate; D000941:Antigens; D015703:Antigens, CD; C558381:BTN3A1 protein, human; C576586:BTN3A2 protein, human; D000071797:Butyrophilins; D011756:Diphosphates; D004164:Diphosphonates; D045782:Hemiterpenes; D007093:Imidazoles; D008565:Membrane Proteins; D009943:Organophosphorus Compounds; C491674:PPL protein, human; D010750:Phosphoproteins; D051179:Plakins; D016692:Receptors, Antigen, T-Cell, gamma-delta; D011993:Recombinant Fusion Proteins; C004809:isopentenyl pyrophosphate; D000077211:Zoledronic Acid
references: Nat Rev Immunol. 2013 Feb;13(2):88-100
references: 23348415
references: J Cell Biol. 2004 Aug 30;166(5):653-9
references: 15337775
references: PLoS One. 2013;8(7):e69767
references: 23922795
references: Nat Immunol. 2013 Sep;14(9):886-7
references: 23959177
references: Nat Immunol. 2013 Sep;14(9):908-16
references: 23872678
references: Nature. 2008 Dec 18;456(7224):976-79
references: 19140243
references: PLoS One. 2009;4(2):e4503
references: 19223974
references: Cell. 2010 Apr 2;141(1):117-28
references: 20371349
references: Eur J Immunol. 2012 Jul;42(7):1668-76
references: 22806069
references: J Cell Biol. 1997 Dec 29;139(7):1835-49
references: 9412476
references: Genomics. 1999 Mar 1;56(2):160-8
references: 10051401
references: J Biol Chem. 2007 Apr 13;282(15):11540-8
references: 17298950
references: Proc Natl Acad Sci U S A. 2007 Apr 10;104(15):6200-5
references: 17400754
references: Traffic. 2007 May;8(5):594-604
references: 17451558
references: J Cell Biol. 2007 Dec 31;179(7):1599-612
references: 18166659
references: Nat Genet. 2008 May;40(5):656-62
references: 18408721
references: Proc Natl Acad Sci U S A. 2012 Aug 14;109(33):13278-83
references: 22847415
references: Blood. 2012 Sep 13;120(11):2269-79
references: 22767497
references: J Biol Chem. 2012 Sep 21;287(39):32780-90
references: 22846996
references: Immunity. 2014 Apr 17;40(4):490-500
references: 24703779
references: Immunol Lett. 2014 Sep;161(1):133-7
references: 24925024
references: Chem Biol. 2014 Aug 14;21(8):945-54
references: 25065532
references: Eur J Immunol. 2014 Sep;44(9):2571-6
references: 24890657
references: J Cell Biol. 2000 Oct 30;151(3):573-86
references: 11062259
references: Genomics. 2001 Feb 1;71(3):351-62
references: 11170752
references: FEBS Lett. 2003 Jun 5;544(1-3):4-10
references: 12782281
references: Nat Rev Mol Cell Biol. 2004 Jul;5(7):542-53
references: 15232572
references: J Immunol. 2013 Aug 1;191(3):1029-42
references: 23833237