title: Fcγ receptor dependency of agonistic CD40 antibody in lymphoma therapy can be overcome through antibody multimerization.
abstract: Immunomodulatory mAbs, led by the anti-CTLA4 mAb ipilimumab, are an exciting new class of drugs capable of promoting anticancer immunity and providing durable control of some tumors. Close analysis of a number of agents has revealed a critical yet variable role for Fcγ receptors in their efficacy. In this article, we reveal that agonistic anti-CD40 mAbs have an absolute requirement for cross-linking by inhibitory FcγRIIB when used systemically to treat established BCL1 syngeneic lymphoma, and therapy is lost when using a mouse IgG2a mAb not cross-linked by FcγRIIB. Furthermore, in FcγRIIB-deficient mice the lymphoma itself can provide FcγRIIB to cross-link anti-CD40 on neighboring cells, and only when this is blocked does therapy fail. The dependence on FcγRIIB for immunostimulatory activity was not absolute, however, because when anti-CD40 mAbs were administered systemically with the TLR3 agonist polyinosinic:polycytidylic acid or were given subcutaneously, activatory FcγR could also provide cross-linking. Using this mechanistic insight, we designed multimeric forms of anti-CD40 mAb with intrinsic FcγR-independent activity that were highly effective in the treatment of lymphoma-bearing mice. In conclusion, FcγR-independent anti-CD40 activation is a viable strategy in vivo. These findings have important translational implications, as humans, unlike mice, do not have IgG that binds strongly to FcγRIIB; therefore FcγR-independent derivatives represent an attractive therapeutic option.
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Ann L
authors.initials: AL
authors.lastname: White
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Lang
authors.initials: L
authors.lastname: Dou
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: H T Claude
authors.initials: HT
authors.lastname: Chan
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Vikki L
authors.initials: VL
authors.lastname: Field
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: C Ian
authors.initials: CI
authors.lastname: Mockridge
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Kane
authors.initials: K
authors.lastname: Moss
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Emily L
authors.initials: EL
authors.lastname: Williams
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Steven G
authors.initials: SG
authors.lastname: Booth
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Ruth R
authors.initials: RR
authors.lastname: French
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Potter
authors.affiliation: Immunology Research, Biogen Idec, Cambridge, MA 02142;
authors.forename: Cherié
authors.initials: C
authors.lastname: Butts
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Aymen
authors.initials: A
authors.lastname: Al-Shamkhani
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Mark S
authors.initials: MS
authors.lastname: Cragg
authors.affiliation: Department of Human Genetics, Leiden University Medical Centre, 2333 ZA Leiden, the Netherlands; and.
authors.forename: J Sjef
authors.initials: JS
authors.lastname: Verbeek
authors.affiliation: Cancer Sciences Unit, Southampton Cancer Research UK Centre, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom.
authors.forename: Peter W M
authors.initials: PW
authors.lastname: Johnson
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom;
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Glennie
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton Faculty of Medicine, Southampton SO16 6YD, United Kingdom; sab@soton.ac.uk.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Beers
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000627:therapeutic use*; D001402:B-Lymphocytes / Q000276:immunology; D019013:CD40 Antigens / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D049109:Cell Proliferation; D002478:Cells, Cultured; D007074:Immunoglobulin G / Q000276:immunology / Q000627:therapeutic use; D007167:Immunotherapy; D008223:Lymphoma / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011070:Poly I-C / Q000627:therapeutic use; D055503:Protein Multimerization / Q000276:immunology*; D017452:Receptors, IgG / Q000235:genetics / Q000276:immunology*; D020349:Surface Plasmon Resonance; D051196:Toll-Like Receptor 3 / Q000819:agonists
chemical_list: D000911:Antibodies, Monoclonal; D019013:CD40 Antigens; C080571:Fc gamma receptor IIB; D007074:Immunoglobulin G; D017452:Receptors, IgG; C519703:TLR3 protein, mouse; D051196:Toll-Like Receptor 3; D011070:Poly I-C