title: Conformation of the human immunoglobulin G2 hinge imparts superagonistic properties to immunostimulatory anticancer antibodies.
abstract: Monoclonal antibody (mAb) drugs that stimulate antitumor immunity are transforming cancer treatment but require optimization for maximum clinical impact. Here, we show that, unlike other immunoglobulin isotypes, human IgG2 (h2) imparts FcγR-independent agonistic activity to immune-stimulatory mAbs such as anti-CD40, -4-1BB, and -CD28. Activity is provided by a subfraction of h2, h2B, that is structurally constrained due its unique arrangement of hinge region disulfide bonds. Agonistic activity can be transferred from h2 to h1 by swapping their hinge and CH1 domains, and substitution of key hinge and CH1 cysteines generates homogenous h2 variants with distinct agonistic properties. This provides the exciting opportunity to engineer clinical reagents with defined therapeutic activity regardless of FcγR expression levels in the local microenvironment.
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK. Electronic address: a.l.white@soton.ac.uk.
authors.forename: Ann L
authors.initials: AL
authors.lastname: White
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: H T Claude
authors.initials: HT
authors.lastname: Chan
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Ruth R
authors.initials: RR
authors.lastname: French
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Jane
authors.initials: J
authors.lastname: Willoughby
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: C Ian
authors.initials: CI
authors.lastname: Mockridge
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Ali
authors.initials: A
authors.lastname: Roghanian
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Christine A
authors.initials: CA
authors.lastname: Penfold
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Steven G
authors.initials: SG
authors.lastname: Booth
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Ali
authors.initials: A
authors.lastname: Dodhy
authors.affiliation: Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Marta E
authors.initials: ME
authors.lastname: Polak
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Potter
authors.affiliation: Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Ardern-Jones
authors.affiliation: Department of Human Genetics, Leiden University Medical Centre, Albinusdreef 2, 2333 ZA Leiden, the Netherlands.
authors.forename: J Sjef
authors.initials: JS
authors.lastname: Verbeek
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Peter W M
authors.initials: PW
authors.lastname: Johnson
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Aymen
authors.initials: A
authors.lastname: Al-Shamkhani
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Cragg
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Beers
authors.affiliation: Cancer Sciences Unit, Faculty of Medicine, University of Southampton, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Glennie
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology / Q000627:therapeutic use*; D018106:CD28 Antigens / Q000276:immunology; D019013:CD40 Antigens / Q000276:immunology; D002478:Cells, Cultured; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology / Q000627:therapeutic use*; D051379:Mice; D008810:Mice, Inbred C57BL; D017452:Receptors, IgG / Q000276:immunology*; D013945:Thymoma / Q000188:drug therapy / Q000276:immunology / Q000517:prevention & control*; D013953:Thymus Neoplasms / Q000188:drug therapy / Q000276:immunology / Q000517:prevention & control*; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000276:immunology; D014611:Vaccination / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D018106:CD28 Antigens; D019013:CD40 Antigens; D007074:Immunoglobulin G; D017452:Receptors, IgG; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9