FcγRIIIb Restricts Antibody-Dependent Destruction of Cancer Cells by Human Neutrophils

Abstract

The function of the low-affinity IgG-receptor FcγRIIIb (CD16b), which is uniquely and abundantly expressed on human granulocytes, is not clear. Unlike the other Fcγ receptors (FcγR), it is a glycophosphatidyl inositol (GPI) -anchored molecule and does not have intracellular signaling motifs. Nevertheless, FcγRIIIb can cooperate with other FcγR to promote phagocytosis of antibody-opsonized microbes by human neutrophils. Here we have investigated the role of FcγRIIIb during antibody-dependent cellular cytotoxicity (ADCC) by neutrophils toward solid cancer cells coated with either trastuzumab (anti-HER2) or cetuximab (anti-EGFR). Inhibiting FcγRIIIb using CD16-F(ab')2 blocking antibodies resulted in substantially enhanced ADCC. ADCC was completely dependent on FcγRIIa (CD32a) and the enhanced ADCC seen after FcγRIIIb blockade therefore suggested that FcγRIIIb was competing with FcγRIIa for IgG on the opsonized target cells. Interestingly, the function of neutrophil FcγRIIIb as a decoy receptor was further supported by using neutrophils from individuals with different gene copy numbers of FCGR3B causing different levels of surface FcγRIIIb expression. Individuals with one copy of FCGR3B showed higher levels of ADCC compared to those with two or more copies. Finally, we show that therapeutic antibodies intended to improve FcγRIIIa (CD16a)-dependent natural killer (NK) cell ADCC due to the lack of fucosylation on the N-linked glycan at position N297 of the IgG1 heavy chain Fc-region, show decreased ADCC as compared to regularly fucosylated antibodies. Together, these data confirm FcγRIIIb as a negative regulator of neutrophil ADCC toward tumor cells and a potential target for enhancing tumor cell destruction by neutrophils.

Redirected optimized cell killing (ROCK®): A highly versatile multispecific fit-for-purpose antibody platform for engaging innate immunity

“…In addition, the contribution of neutrophils, the most abundant type of granulocytes, and the role of CD16B in tumor control is not fully understood. 78,79 Therefore, it is suggested that also those patients with a low affinity CD16A genotype (158F, roughly 85% of the Caucasian population) 74,75 will respond better to immune cell engagers based on the ROCK® anti-CD16A domains. Moreover, the lack of binding to Fc receptors other than CD16A could substantially reduce sink-effects compared to conventional mAbs.…”

IgG-aggregates rapidly up-regulate FcgRI expression at the surface of human neutrophils in a FcgRII-dependent fashion: A crucial role for FcgRI in the generation of reactive oxygen species

“…The FcgRIIIb-specific 11.5 blocking antibody, actually enhanced FcgRI up-regulation, as did PI-PLC, a FcgRIIIb-specific shedding enzyme. Moreover, the 3G8 blocking antibody (36), which does not discriminate between the two FcgRIII isoforms, did not affect FcgRI up-regulation, supporting the concept of a decoy role for that receptor (10,37,38). Results leave the possibility of a role for FcgRIIIa.…”

IgG‐aggregates rapidly upregulate FcgRI expression at the surface of human neutrophils in a FcgRII‐dependent fashion: A crucial role for FcgRI in the generation of reactive oxygen species

“…The FcgRIIIb-specific 11.5 blocking antibody, actually enhanced FcgRI upregulation, as did PI-PLC, a FcgRIIIb-specific shedding enzyme. Moreover, the 3G8 blocking antibody, 57 which does not discriminate between the two FcgRIII isoforms, did not affect FcgRI upregulation, supporting the concept of a decoy role for FcgRIIIb 12,58,59 in this process. Results leave the possibility of a role for FcgRIIIa.…”