title: Immunotherapy targeting inhibitory Fcγ receptor IIB (CD32b) in the mouse is limited by monoclonal antibody consumption and receptor internalization.
abstract: Genetic deficiency of the inhibitory Fc receptor, FcγRIIB (CD32b), has been shown to augment the activity of activatory FcγR and promote mAb immunotherapy. To investigate whether mAbs capable of blocking FcγRIIB have similar capacity, we recently generated a panel of specific anti-mouse FcγRIIB mAbs that do not cross-react with other FcRs, allowing us to study the potential of FcγRIIB as a therapeutic target. Previous work revealed a number of these mAbs capable of eliciting programmed cell death of targets, and in the present study we demonstrated their ability to promote target cell phagocytosis. However, in a variety of murine tumor models, anti-FcγRIIB mAbs demonstrated limited therapeutic activity despite optimized treatment regimens. Unexpectedly, we observed that the anti-FcγRIIB mAbs are rapidly and extensively consumed in vivo, both by the tumor and host cells, including B cells, leading to a precipitous loss from the circulation. Closer analysis revealed that the anti-FcγRIIB mAbs become extensively internalized from the cell surface within 24 h in vivo, likely explaining their suboptimal efficacy. Subsequent studies revealed that anti-FcγRIIB mAb immunotherapy was effective when used against FcγRIIB(+) tumors in FcγRIIB(-/-) recipients, indicating that consumption of the mAb by nontumor cells is the primary limitation of these reagents. Importantly, similar rates of internalization were not seen on human target cells, at least in vitro. These studies further highlight the need to determine the propensity of mAb therapeutics to internalize target receptors and also identify potential key differences between human and mouse cells in this respect.
authors.affiliation: Antibody and Vaccine Group, Cancer Sciences Unit, University of Southampton, Faculty of Medicine, General Hospital, Southampton SO16 6YD, United Kingdom;
authors.forename: Emily L
authors.initials: EL
authors.lastname: Williams
authors.forename: Alison L
authors.initials: AL
authors.lastname: Tutt
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Beers
authors.forename: Ruth R
authors.initials: RR
authors.lastname: French
authors.forename: Claude H T
authors.initials: CH
authors.lastname: Chan
authors.forename: Kerry L
authors.initials: KL
authors.lastname: Cox
authors.forename: Ali
authors.initials: A
authors.lastname: Roghanian
authors.forename: Christine A
authors.initials: CA
authors.lastname: Penfold
authors.forename: Cherié L
authors.initials: CL
authors.lastname: Butts
authors.forename: Peter
authors.initials: P
authors.lastname: Boross
authors.forename: J Sjef
authors.initials: JS
authors.lastname: Verbeek
authors.forename: Mark S
authors.initials: MS
authors.lastname: Cragg
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Glennie
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D017209:Apoptosis / Q000276:immunology; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D007167:Immunotherapy; D016393:Lymphoma, B-Cell / Q000276:immunology* / Q000628:therapy; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D018345:Mice, Knockout; D009101:Multiple Myeloma / Q000276:immunology* / Q000628:therapy; D017452:Receptors, IgG / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; C080571:Fc gamma receptor IIB; D017452:Receptors, IgG