title: Highly specific and potently activating Vγ9Vδ2-T cell specific nanobodies for diagnostic and therapeutic applications.
abstract: Vγ9Vδ2-T cells constitute the predominant subset of γδ-T cells in human peripheral blood and have been shown to play an important role in antimicrobial and antitumor immune responses. Several efforts have been initiated to exploit these cells for cancer immunotherapy, e.g. by using phosphoantigens, adoptive cell transfer, and by a bispecific monoclonal antibody based approach. Here, we report the generation of a novel set of Vγ9Vδ2-T cell specific VHH (or nanobody). VHH have several advantages compared to conventional antibodies related to their small size, stability, ease of generating multispecific molecules and low immunogenicity. With high specificity and affinity, the anti-Vγ9Vδ2-T cell receptor VHHs are shown to be useful for FACS, MACS and immunocytochemistry. In addition, some VHH were found to specifically activate Vγ9Vδ2-T cells. Besides being of possible immunotherapeutic value, these single domain antibodies will be of great value in the further study of this important immune effector cell subset.
authors.affiliation: Department of Medical Oncology, VU University Medical Center, De Boelelaan 1117, 1081, HV, Amsterdam, The Netherlands. Electronic address: rc.debruin@vumc.nl.
authors.forename: Renée C G
authors.initials: RCG
authors.lastname: de Bruin
authors.affiliation: Department of Medical Oncology, VU University Medical Center, De Boelelaan 1117, 1081, HV, Amsterdam, The Netherlands. Electronic address: sm.lougheed@vumc.nl.
authors.forename: Sinéad M
authors.initials: SM
authors.lastname: Lougheed
authors.affiliation: Department of Medical Oncology, VU University Medical Center, De Boelelaan 1117, 1081, HV, Amsterdam, The Netherlands. Electronic address: kruk_lizz@hotmail.com.
authors.forename: Liza
authors.initials: L
authors.lastname: van der Kruk
authors.affiliation: Department of Medical Oncology, VU University Medical Center, De Boelelaan 1117, 1081, HV, Amsterdam, The Netherlands. Electronic address: agm.stam@vumc.nl.
authors.forename: Anita G
authors.initials: AG
authors.lastname: Stam
authors.affiliation: Department of Pathology, VU University Medical Center, De Boelelaan 1117, 1081, HV, Amsterdam, The Netherlands. Electronic address: erik.hooijberg@vumc.nl.
authors.forename: Erik
authors.initials: E
authors.lastname: Hooijberg
authors.affiliation: Department of Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands. Electronic address: r.roovers@merus.nl.
authors.forename: Rob C
authors.initials: RC
authors.lastname: Roovers
authors.affiliation: Department of Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands. Electronic address: p.vanbergen@uu.nl.
authors.forename: Paul M P
authors.initials: PMP
authors.lastname: van Bergen En Henegouwen
authors.affiliation: Department of Medical Oncology, VU University Medical Center, De Boelelaan 1117, 1081, HV, Amsterdam, The Netherlands. Electronic address: h.verheul@vumc.nl.
authors.forename: Henk M W
authors.initials: HMW
authors.lastname: Verheul
authors.affiliation: Department of Medical Oncology, VU University Medical Center, De Boelelaan 1117, 1081, HV, Amsterdam, The Netherlands. Electronic address: td.degruijl@vumc.nl.
authors.forename: Tanja D
authors.initials: TD
authors.lastname: de Gruijl
authors.affiliation: Department of Medical Oncology, VU University Medical Center, De Boelelaan 1117, 1081, HV, Amsterdam, The Netherlands. Electronic address: jj.vandervliet@vumc.nl.
authors.forename: Hans J
authors.initials: HJ
authors.lastname: van der Vliet
mesh_terms: D000818:Animals; D000915:Antibody Affinity / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D002161:Camelids, New World / Q000276:immunology; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D018189:Immunomagnetic Separation / Q000379:methods; D019169:Jurkat Cells; D008856:Microscopy, Fluorescence; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000276:immunology* / Q000378:metabolism; D015203:Reproducibility of Results; D061905:Single-Domain Antibodies / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
chemical_list: D016692:Receptors, Antigen, T-Cell, gamma-delta; D061905:Single-Domain Antibodies
keywords: Cancer; Flow cytometry; Gamma delta T cells; Immunocytochemistry; Immunotherapy; MACS; Nanobody; Single-domain antibody fragment; VHH