Avidity confers FcγR binding and immune effector function to aglycosylated immunoglobulin G1

Nesspor, Thomas; Raju, T. Shantha; Chin, Chen Ni; Vafa, Omid; Brezski, Randall J.

doi:10.1002/jmr.2155

Cited by 49 publications

(40 citation statements)

References 36 publications

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“…S4). This activation suggests that for this application, a (bs)IgG1-N297Q backbone may not be silent enough, as also demonstrated by others (43), and alternative nonactivating backbones should be evaluated.…”

Section: Discussionmentioning

confidence: 56%

Efficient generation of stable bispecific IgG1 by controlled Fab-arm exchange

Labrijn

Meesters

Goeij

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

274

298

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The promise of bispecific antibodies (bsAbs) to yield more effective therapeutics is well recognized; however, the generation of bsAbs in a practical and cost-effective manner has been a formidable challenge. Here we present a technology for the efficient generation of bsAbs with normal IgG structures that is amenable to both antibody drug discovery and development. The process involves separate expression of two parental antibodies, each containing single matched point mutations in the CH3 domains. The parental antibodies are mixed and subjected to controlled reducing conditions in vitro that separate the antibodies into HL half-molecules and allow reassembly and reoxidation to form highly pure bsAbs. The technology is compatible with standard large-scale antibody manufacturing and ensures bsAbs with Fcmediated effector functions and in vivo stability typical of IgG1 antibodies. Proof-of-concept studies with HER2×CD3 (T-cell recruitment) and HER2×HER2 (dual epitope targeting) bsAbs demonstrate superior in vivo activity compared with parental antibody pairs.immunotherapy | pharmacokinetics | anti-tumor

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Section: Discussionmentioning

confidence: 56%

Efficient generation of stable bispecific IgG1 by controlled Fab-arm exchange

Labrijn

Meesters

Goeij

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

274

298

View full text Add to dashboard Cite

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“…Additionally, unlike PBMC-mediated ADCC, multiple IgG subclasses can facilitate macrophage-mediated ADCP. 16,28,46 We have previously reported a macrophage-mediated tumor cell-killing assay that assesses the total loss of tumor cells over a 24-h period as opposed to the more commonly employed 4-h assessments of macrophage internalization of mAb-opsonized tumor cells. 28 For the purposes of this study, we performed 24-h macrophage-mediated cell-killing assays and also collected supernatants to assess the levels of IL-10 at the 24-h time point.…”

Section: Resultsmentioning

confidence: 99%

An Fc engineering approach that modulates antibody-dependent cytokine release without altering cell-killing functions

Kinder

Greenplate

Strohl

et al. 2015

mAbs

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(2015) An Fc engineering approach that modulates antibodydependent cytokine release without altering cell-killing functions, mAbs, 7:3, 494-504, DOI: 10.1080DOI: 10. /19420862.2015 To link to this article: https://doi.org/10. 1080/19420862.2015 Cytotoxic therapeutic monoclonal antibodies (mAbs) often mediate target cell-killing by eliciting immune effector functions via Fc region interactions with cellular and humoral components of the immune system. Key functions include antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and complement-dependent cytotoxicity (CDC). However, there has been increased appreciation that along with cell-killing functions, the induction of antibody-dependent cytokine release (ADCR) can also influence disease microenvironments and therapeutic outcomes. Historically, most Fc engineering approaches have been aimed toward modulating ADCC, ADCP, or CDC. In the present study, we describe an Fc engineering approach that, while not resulting in impaired ADCC or ADCP, profoundly affects ADCR. As such, when peripheral blood mononuclear cells are used as effector cells against mAb-opsonized tumor cells, the described mAb variants elicit a similar profile and quantity of cytokines as IgG1. In contrast, although the variants elicit similar levels of tumor cell-killing as IgG1 with macrophage effector cells, the variants do not elicit macrophage-mediated ADCR against mAb-opsonized tumor cells. This study demonstrates that Fc engineering approaches can be employed to uncouple macrophage-mediated phagocytic and subsequent cell-killing functions from cytokine release.

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“…However, the physiological interaction of IgG immune complexes (ICs) and FcgR requires avid binding of the complex through the display of multiple Fc regions of "near-neighbor" IgGs to engage and cluster multiple FcgRs on the cell surface (36). Hence, differences in the opsonization of targets by different IgGs influence interactions with FcgRs, chiefly by the density, size (37,38), and topology of presentation of the Fc regions.…”

mentioning

confidence: 99%

Dimeric FcγR Ectodomains as Probes of the Fc Receptor Function of Anti-Influenza Virus IgG

Wines

Vanderven

Esparon

et al. 2016

The Journal of Immunology

130

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Ab-dependent cellular cytotoxicity, phagocytosis, and Ag presentation are key mechanisms of action of Abs arising in vaccine or naturally acquired immunity, as well of therapeutic mAbs. Cells expressing the low-affinity FcγRs (FcγRII or CD32 and FcγRIII or CD16) are activated for these functions when receptors are aggregated following the binding of IgG-opsonized targets. Despite the diversity of the Fc receptor proteins, IgG ligands, and potential responding cell types, the induction of all FcγR-mediated responses by opsonized targets requires the presentation of multiple Fc regions in close proximity to each other. We demonstrated that such “near-neighbor” Fc regions can be detected using defined recombinant soluble (rs) dimeric low-affinity ectodomains (rsFcγR) that have an absolute binding requirement for the simultaneous engagement of two IgG Fc regions. Like cell surface–expressed FcγRs, the binding of dimeric rsFcγR ectodomains to Ab immune complexes was affected by Ab subclass, presentation, opsonization density, Fc fucosylation, or mutation. The activation of an NK cell line and primary NK cells by human IgG-opsonized influenza A hemagglutinin correlated with dimeric rsFcγRIIIa binding activity but not with Ab titer. Furthermore, the dimeric rsFcγR binding assay sensitively detected greater Fc receptor activity to pandemic H1N1 hemagglutinin after the swine influenza pandemic of 2009 in pooled human polyclonal IgG. Thus these dimeric rsFcγR ectodomains are validated, defined probes that should prove valuable in measuring the immune-activating capacity of IgG Abs elicited by infection or vaccination or experimentally derived IgG and its variants.

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Avidity confers FcγR binding and immune effector function to aglycosylated immunoglobulin G1

Cited by 49 publications

References 36 publications

Efficient generation of stable bispecific IgG1 by controlled Fab-arm exchange

Efficient generation of stable bispecific IgG1 by controlled Fab-arm exchange

An Fc engineering approach that modulates antibody-dependent cytokine release without altering cell-killing functions

Dimeric FcγR Ectodomains as Probes of the Fc Receptor Function of Anti-Influenza Virus IgG

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