title: RhoB Mediates Phosphoantigen Recognition by Vγ9Vδ2 T Cell Receptor.
abstract: Human Vγ9Vδ2 T cells respond to tumor cells by sensing elevated levels of phosphorylated intermediates of the dysregulated mevalonate pathway, which is translated into activating signals by the ubiquitously expressed butyrophilin A1 (BTN3A1) through yet unknown mechanisms. Here, we developed an unbiased, genome-wide screening method that identified RhoB as a critical mediator of Vγ9Vδ2 TCR activation in tumor cells. Our results show that Vγ9Vδ2 TCR activation is modulated by the GTPase activity of RhoB and its redistribution to BTN3A1. This is associated with cytoskeletal changes that directly stabilize BTN3A1 in the membrane, and the subsequent dissociation of RhoB from BTN3A1. Furthermore, phosphoantigen accumulation induces a conformational change in BTN3A1, rendering its extracellular domains recognizable by Vγ9Vδ2 TCRs. These complementary events provide further evidence for inside-out signaling as an essential step in the recognition of tumor cells by a Vγ9Vδ2 TCR.
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Sebestyen
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands.
authors.forename: Wouter
authors.initials: W
authors.lastname: Scheper
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands.
authors.forename: Anna
authors.initials: A
authors.lastname: Vyborova
authors.affiliation: Department of Clinical Chemistry and Hematology, University Medical Center, Utrecht 3508 GA, the Netherlands.
authors.forename: Siyi
authors.initials: S
authors.lastname: Gu
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Rychnavska
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands.
authors.forename: Marleen
authors.initials: M
authors.lastname: Schiffler
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands.
authors.forename: Astrid
authors.initials: A
authors.lastname: Cleven
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands.
authors.forename: Coraline
authors.initials: C
authors.lastname: Chéneau
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands.
authors.forename: Martje
authors.initials: M
authors.lastname: van Noorden
authors.affiliation: 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.
authors.forename: Cassie-Marie
authors.initials: CM
authors.lastname: Peigné
authors.affiliation: INSERM, Centre de Recherche en Cancérologie Marseille, Institut Paoli-Calmettes, 13009 Marseille, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Olive
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, Utrecht 3584, the Netherlands.
authors.forename: Robert Jan
authors.initials: RJ
authors.lastname: Lebbink
authors.affiliation: Department of Clinical Chemistry and Hematology, University Medical Center, Utrecht 3508 GA, the Netherlands.
authors.forename: Rimke
authors.initials: R
authors.lastname: Oostvogels
authors.affiliation: Department of Clinical Chemistry and Hematology, University Medical Center, Utrecht 3508 GA, the Netherlands.
authors.forename: Tuna
authors.initials: T
authors.lastname: Mutis
authors.affiliation: Department of Hematology, VU University Medical Center, Amsterdam 1081, the Netherlands.
authors.forename: Gerrit Jan
authors.initials: GJ
authors.lastname: Schuurhuis
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Chicago, 929 East 57(th) Street, Chicago, IL 60615, USA.
authors.forename: Erin J
authors.initials: EJ
authors.lastname: Adams
authors.affiliation: 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.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Scotet
authors.affiliation: Department of Hematology and Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht 3508, the Netherlands. Electronic address: j.h.e.kuball@umcutrecht.nl.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Kuball
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000941:Antigens / Q000378:metabolism; D015703:Antigens, CD / Q000737:chemistry / Q000378:metabolism; D000071797:Butyrophilins / Q000737:chemistry / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D056426:Genetic Loci; D057809:HEK293 Cells; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D008954:Models, Biological; D014411:Neoplastic Stem Cells / Q000378:metabolism; D010766:Phosphorylation; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D055503:Protein Multimerization; D034741:RNA, Small Interfering / Q000378:metabolism; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000378:metabolism*; D020743:rhoB GTP-Binding Protein / Q000378:metabolism*
chemical_list: D000941:Antigens; D015703:Antigens, CD; C558381:BTN3A1 protein, human; D000071797:Butyrophilins; D034741:RNA, Small Interfering; D016692:Receptors, Antigen, T-Cell, gamma-delta; D020743:rhoB GTP-Binding Protein