Dataset Information

Antibody to Fc?RI? Suppresses Immunoglobulin E Binding to High-Affinity Receptor I in Allergic Inflammation.

ABSTRACT:

Purpose

High-affinity receptor I (Fc?RI) on mast cells and basophils plays a key role in the immunoglobulin E (IgE)-mediated type I hypersensitivity mediated by allergen cross-linking of the specific IgE-Fc?RI complex. Thus, prevention of IgE binding to Fc?RI on these cells is an effective therapy for allergic disease. We have developed a strategy to disrupt IgE binding to Fc?RI using an antibody targeting Fc?RI?.

Materials and methods

Fab fragment antibodies, which lack the Fc domain, with high affinity and specificity for Fc?RI? and effective inhibitory activity against IgE-Fc?RI binding were screened. IgE-induced histamine, ?-hexosaminidase and Ca²? release in basophils were determined by ELISA. A B6.Cg-Fcer1a(tm1Knt) Tg(FCER1A)1Bhk/J mouse model of passive cutaneous anaphylaxis (PCA) was used to examine the inhibitory effect of NPB311 on allergic skin inflammation.

Results

NPB311 exhibited high affinity to human Fc?RI? (KD=4 nM) and inhibited histamine, ?-hexosaminidase and Ca²? release in a concentration-dependent manner in hFc?RI-expressing cells. In hFc?RI?-expressing mice, dye leakage was higher in the PCA group than in controls, but decreased after NPB311 treatment. NPB311 could form a complex with Fc?RI? and inhibit the release of inflammation mediators.

Conclusion

Our approach for producing anti-Fc?RI? Fab fragment antibody NPB311 may enable clinical application to a therapeutic pathway in IgE/Fc?RI-mediated diseases.

SUBMITTER: Hong JY

PROVIDER: S-EPMC5011273 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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Publications

Antibody to FcεRIα Suppresses Immunoglobulin E Binding to High-Affinity Receptor I in Allergic Inflammation.

Hong Jung Yeon JY Bae Jong Hwan JH Lee Kyung Eun KE Kim Mina M Kim Min Hee MH Kang Hyun Jung HJ Park Eun Hye EH Yoo Kyung Sook KS Jeong Se Kyoo SK Kim Kyung Won KW Kim Kyu Earn KE Sohn Myung Hyun MH

Yonsei medical journal 20161101 6

<h4>Purpose</h4>High-affinity receptor I (FcεRI) on mast cells and basophils plays a key role in the immunoglobulin E (IgE)-mediated type I hypersensitivity mediated by allergen cross-linking of the specific IgE-FcεRI complex. Thus, prevention of IgE binding to FcεRI on these cells is an effective therapy for allergic disease. We have developed a strategy to disrupt IgE binding to FcεRI using an antibody targeting FcεRIα.<h4>Materials and methods</h4>Fab fragment antibodies, which lack the Fc do ...[more]

PMID: 27593869

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Project description:Structurally well-defined IgG-Fc glycoforms are highly demanded for understanding the effects of glycosylation on an antibody's effector functions. We report in this paper chemoenzymatic synthesis and Fc? receptor binding of an array of homogeneous IgG-Fc glycoforms. The chemoenzymatic approach consists of the chemical synthesis of defined N-glycan oxazolines as donor substrates, the expression of the Fc domain in a CHO cell line in the presence of an ?-mannosidase inhibitor kifunensine, and an endoglycosidase-catalyzed glycosylation of the deglycosylated Fc domain (GlcNAc-Fc homodimer) with the synthetic glycan oxazolines. The enzyme from Arthrobacter protophormiae (Endo-A) was found to be remarkably efficient to take various modified N-glycan core oxazolines, including the bisecting sugar-containing derivatives, for Fc glycosylation remodeling, resulting in the formation of the corresponding homogeneous Fc glycoforms. Nevertheless, neither Endo-A nor the Mucor hiemalis endoglycosidase mutants (EndoM-N175A and EndoM-N175Q) were able to transfer full-length complex-type N-glycan to the Fc domain, implicating the limitations of these two enzymes in Fc glycosylation remodeling. Surface plasmon resonance (SPR) binding studies with the synthetic IgG-Fc glycoforms unambiguously proved that the presence of a bisecting GlcNAc moiety could significantly enhance the binding of Fc to Fc?RIIIa, the activating Fc? receptor, independent of Fc core-fucosylation. Interestingly, the Fc glycoforms carrying an unusual bisecting sugar moiety such as a mannose or a LacNAc moiety also demonstrated enhanced affinity to Fc?RIIIa. On the orther hand, the presence of a bisecting GlcNAc or core-fucosylation had little effect on the affinity of Fc to the inhibitory Fc? receptor, Fc?RIIb. Our experimental data also showed that the ?-linked mannose residues in the pentasaccharide Man3GlcNAc2 core was essential to maintain a high affinity of Fc to both Fc?RIIIa and Fc?RIIb. The synthetic homogeneous Fc glycoforms thus provide a useful tool for elucidating how a fine Fc N-glycan structure precisely affects the function of the Fc domain.