Engineered hexavalent Fc proteins with enhanced Fc-gamma receptor avidity provide insights into immune-complex interactions

Rowley, Tania F.; Peters, Shirley J.; Aylott, Mike; Griffin, Robert; Davies, Nicola L.; Healy, Louise; Cutler, Rona M.; Eddleston, AdrianL.W.F.; Pither, Thomas Leonard; Sopp, Joshua M.; Zaccheo, Oliver; Fossati, Gianluca; Cain, Katharine; Ventom, Andrew M.; Hailu, Hanna; Ward, Emily M. Geraghty; Sherington, John; Brennan, Frank R.; Fallah-Arani, Farnaz; Humphreys, David P.

doi:10.1038/s42003-018-0149-9

Cited by 20 publications

(27 citation statements)

References 71 publications

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“…If any lessons are to be learned, it is that each antibody must be thoroughly evaluated using systems that recapitulate as closely as possible endogenous immune system components. One striking example of this tenet is the observation that the efficacy of a hexameric IgG1 Fc increased when neutrophils and platelets were incorporated in an in-vitro assay with PBMCs (165). Moreover, soluble complement components can also bind the immune complex to affect the immune response as reported in few studies described above (130, 148, 149).…”

Section: Discussionmentioning

confidence: 99%

“…The use of different IgG subclasses in designed immune complexes can also impact potential therapeutic use. Incubation of a hexameric IgG1 Fc construct, discussed above as an inhibitor of phagocytosis, elicited the release of higher cytokine levels in whole blood when compared to PBMCs, likely due to CD16b engagement on neutrophils (not present in PBMCs) (165). Furthermore, the hexameric IgG1 Fc construct also triggered release of cytokines from platelets through a CD32a-dependent interaction.…”

Section: How Multivalency Impacts Igg-fcγr Interactionsmentioning

confidence: 99%

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Multiple Variables at the Leukocyte Cell Surface Impact Fc γ Receptor-Dependent Mechanisms

2019

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Fc γ receptors (FcγR) expressed on the surface of human leukocytes bind clusters of immunoglobulin G (IgG) to induce a variety of responses. Many therapeutic antibodies and vaccine-elicited antibodies prevent or treat infectious diseases, cancers and autoimmune disorders by binding FcγRs, thus there is a need to fully define the variables that impact antibody-induced mechanisms to properly evaluate candidate therapies and design new intervention strategies. A multitude of factors influence the IgG-FcγR interaction; one well-described factor is the differential affinity of the six distinct FcγRs for the four human IgG subclasses. However, there are several other recently described factors that may prove more relevant for disease treatment. This review covers recent reports of several aspects found at the leukocyte membrane or outside the cell that contribute to the cell-based response to antibody-coated targets. One major focus is recent reports covering post-translational modification of the FcγRs, including asparagine-linked glycosylation. This review also covers the organization of FcγRs at the cell surface, and properties of the immune complex. Recent technical advances provide high-resolution measurements of these often-overlooked variables in leukocyte function and immune system activation.

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Section: Discussionmentioning

confidence: 99%

Section: How Multivalency Impacts Igg-fcγr Interactionsmentioning

confidence: 99%

Multiple Variables at the Leukocyte Cell Surface Impact Fc γ Receptor-Dependent Mechanisms

2019

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“…Many groups have postulated that multivalent Fc constructs have potential for the treatment of immune conditions involving pathogenic Abs (2, 5, 37, 38), and a recent study has shown that hexavalent Fcs can block FcγRs leading to their downmodulation and prolonged disruption of FcγR effector functions both in vitro and in vivo (39, 40). Hexameric Fcs have also been shown to inhibit platelet phagocytosis in mouse models of idiopathic thrombocytopenic purpura (33, 39, 41).…”

Section: Discussionmentioning

confidence: 99%

Insertion of N-Terminal Hinge Glycosylation Enhances Interactions of the Fc Region of Human IgG1 Monomers with Glycan-Dependent Receptors and Blocks Hemagglutination by the Influenza Virus

Blundell

Wilkinson

et al. 2019

The Journal of Immunology

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In therapeutic applications in which the Fc of IgG is critically important, the receptor binding and functional properties of the Fc are lost after deglycosylation or removal of the unique Asn 297 N-X-(T/S) sequon. A population of Fcs bearing sialylated glycans has been identified as contributing to this functionality, and high levels of sialylation also lead to longer serum retention times advantageous for therapy. The efficacy of sialylated Fc has generated an incentive to modify the unique N-linked glycosylation site at Asn 297 , either through chemical and enzymatic methods or by mutagenesis of the Fc, that disrupts the protein–Asn 297 carbohydrate interface. In this study, we took an alternative approach by inserting or deleting N-linked attachment sites into the body of the Fc to generate a portfolio of mutants with tailored effector functions. For example, we describe mutants with enhanced binding to low-affinity inhibitory human Fcγ and glycan receptors that may be usefully incorporated into existing Ab engineering approaches to treat or vaccinate against disease. The IgG1 Fc fragments containing complex sialylated glycans attached to the N-terminal Asn 221 sequon bound influenza virus hemagglutinin and disrupted influenza A–mediated agglutination of human erythrocytes.

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“…The ADCC and CDC activities of the anti-OX40 antibody with the E345R mutation were affected by the choice of IgG subtypes [362]. With so many oligomeric Ab targets, there are continuing applications of hexameric therapeutic Abs that can affect downstream signaling events [328,329,363].…”

Section: Avidity Modulationmentioning

confidence: 99%

Antibody Structure and Function: The Basis for Engineering Therapeutics

et al. 2019

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Antibodies and antibody-derived macromolecules have established themselves as the mainstay in protein-based therapeutic molecules (biologics). Our knowledge of the structure–function relationships of antibodies provides a platform for protein engineering that has been exploited to generate a wide range of biologics for a host of therapeutic indications. In this review, our basic understanding of the antibody structure is described along with how that knowledge has leveraged the engineering of antibody and antibody-related therapeutics having the appropriate antigen affinity, effector function, and biophysical properties. The platforms examined include the development of antibodies, antibody fragments, bispecific antibody, and antibody fusion products, whose efficacy and manufacturability can be improved via humanization, affinity modulation, and stability enhancement. We also review the design and selection of binding arms, and avidity modulation. Different strategies of preparing bispecific and multispecific molecules for an array of therapeutic applications are included.

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Engineered hexavalent Fc proteins with enhanced Fc-gamma receptor avidity provide insights into immune-complex interactions

Cited by 20 publications

References 71 publications

Multiple Variables at the Leukocyte Cell Surface Impact Fc γ Receptor-Dependent Mechanisms

Multiple Variables at the Leukocyte Cell Surface Impact Fc γ Receptor-Dependent Mechanisms

Insertion of N-Terminal Hinge Glycosylation Enhances Interactions of the Fc Region of Human IgG1 Monomers with Glycan-Dependent Receptors and Blocks Hemagglutination by the Influenza Virus

Antibody Structure and Function: The Basis for Engineering Therapeutics

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