The intracellular B30.2 domain of butyrophilin 3A1 binds phosphoantigens to mediate activation of human Vγ9Vδ2 T cells.
In humans, Vγ9Vδ2 T cells detect tumor cells and microbial infections, including Mycobacterium tuberculosis, through recognition of small pyrophosphate containing organic molecules known as phosphoantigens (pAgs). Key to pAg-mediated activation of Vγ9Vδ2 T cells is the butyrophilin 3A1 (BTN3A1) protein that contains an intracellular B30.2 domain critical to pAg reactivity. Here, we have demonstrated through structural, biophysical, and functional approaches that the intracellular B30.2 domain of BTN3A1 directly binds pAg through a positively charged surface pocket. Charge reversal of pocket residues abrogates binding and Vγ9Vδ2 T cell activation. We have also identified a gain-of-function mutation within this pocket that, when introduced into the B30.2 domain of the nonstimulatory BTN3A3 isoform, transfers pAg binding ability and Vγ9Vδ2 T cell activation. These studies demonstrate that internal sensing of changes in pAg metabolite concentrations by BTN3A1 molecules is a critical step in Vγ9Vδ2 T cell detection of infection and tumorigenesis.
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA.
Andrew
A
Sandstrom
INSERM, Unité Mixte de Recherche 892, Centre de Recherche en Cancérologie Nantes Angers, 44000 Nantes, France; University of Nantes, 44000 Nantes, France; Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 6299, 44000 Nantes, France.
Cassie-Marie
CM
Peigné
INSERM, Unité Mixte de Recherche 892, Centre de Recherche en Cancérologie Nantes Angers, 44000 Nantes, France; University of Nantes, 44000 Nantes, France; Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 6299, 44000 Nantes, France.
Alexandra
A
Léger
Program in Biophysical Sciences, University of Chicago, Chicago, IL 60637, USA.
James E
JE
Crooks
INSERM, Unité Mixte de Recherche 892, Centre de Recherche en Cancérologie Nantes Angers, 44000 Nantes, France; University of Nantes, 44000 Nantes, France; Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 6299, 44000 Nantes, France.
Fabienne
F
Konczak
INSERM, Unité Mixte de Recherche 892, Centre de Recherche en Cancérologie Nantes Angers, 44000 Nantes, France; University of Nantes, 44000 Nantes, France; Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 6299, 44000 Nantes, France.
Marie-Claude
MC
Gesnel
INSERM, Unité Mixte de Recherche 892, Centre de Recherche en Cancérologie Nantes Angers, 44000 Nantes, France; University of Nantes, 44000 Nantes, France; Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 6299, 44000 Nantes, France.
Richard
R
Breathnach
INSERM, Unité Mixte de Recherche 892, Centre de Recherche en Cancérologie Nantes Angers, 44000 Nantes, France; University of Nantes, 44000 Nantes, France; Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 6299, 44000 Nantes, France.
Marc
M
Bonneville
INSERM, Unité Mixte de Recherche 892, Centre de Recherche en Cancérologie Nantes Angers, 44000 Nantes, France; University of Nantes, 44000 Nantes, France; Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 6299, 44000 Nantes, France. Electronic address: emmanuel.scotet@univ-nantes.fr.
Emmanuel
E
Scotet
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA; Committee on Immunology, University of Chicago, Chicago, IL 60637, USA. Electronic address: ejadams@uchicago.edu.
Erin J
EJ
Adams
D000941:Antigens / Q000276:immunology; D015703:Antigens, CD / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000071797:Butyrophilins; D002478:Cells, Cultured; D004164:Diphosphonates / Q000276:immunology; D006801:Humans; D007093:Imidazoles / Q000276:immunology; D042541:Intracellular Space; D008213:Lymphocyte Activation / Q000235:genetics; D009154:Mutation / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D015202:Protein Engineering; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017434:Protein Structure, Tertiary / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology*; D000077211:Zoledronic Acid
D000941:Antigens; D015703:Antigens, CD; C558381:BTN3A1 protein, human; D000071797:Butyrophilins; D004164:Diphosphonates; D007093:Imidazoles; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D016692:Receptors, Antigen, T-Cell, gamma-delta; C521523:T-cell receptor Vdelta2, human; C521522:T-cell receptor Vgamma9, human; D000077211:Zoledronic Acid