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A novel human immunoglobulin Fcγ–Fcɛ bifunctional fusion protein inhibits FcɛRI-mediated degranulation

Nature Medicine volume 8, pages 518–521 (2002)Cite this article

Abstract

Human mast cells and basophils that express the high-affinity immunoglobulin E (IgE) receptor, Fcɛ receptor 1 (FcɛRI), have key roles in allergic diseases. FcɛRI cross-linking stimulates the release of allergic mediators1. Mast cells and basophils co-express FcγRIIb, a low affinity receptor containing an immunoreceptor tyrosine-based inhibitory motif and whose co-aggregation with FcɛRI can block FcɛRI-mediated reactivity2,3,4. Here we designed, expressed and tested the human basophil and mast-cell inhibitory function of a novel chimeric fusion protein, whose structure is γHinge-CHγ2-CHγ3-15aa linker-CHɛ2-CHɛ3-CHɛ4. This Fcγ–Fcɛ fusion protein was expressed as the predicted 140-κD dimer that reacted with anti-human ɛ- and γ-chain specific antibodies. Fcγ–Fcɛ bound to both human FcɛRI and FcγRII. It also showed dose- and time-dependent inhibition of antigen-driven IgE-mediated histamine release from fresh human basophils sensitized with IgE directed against NIP (4-hydroxy-3-iodo-5-nitrophenylacetyl). This was associated with altered Syk signaling. The fusion protein also showed increased inhibition of human anti-NP (4-hydroxy-3-nitrophenylacetyl) and anti-dansyl IgE-mediated passive cutaneous anaphylaxis in transgenic mice expressing human FcɛRIα. Our results show that this chimeric protein is able to form complexes with both FcɛRI and FcγRII, and inhibit mast-cell and basophil function. This approach, using a Fcγ–Fcɛ fusion protein to co-aggregate FcɛRI with a receptor containing an immunoreceptor tyrosine-based inhibition motif, has therapeutic potential in IgE- and FcɛRI-mediated diseases.

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Figure 1: Dose- and time- dependent inhibition of basophil histamine release using GE2.
Figure 2: Co-aggregation of FcγRII and FcɛR1 by GE2 inhibits FcɛRI-mediated Syk phosphorylation.
Figure 3: In vivo immunoglobulin Fcγ-Fcɛ fusion protein inhibits IgE-mediated degranulation in transgenic mice.

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Acknowledgements

We thank S.L. Morrison and R. Trinh for technical assistance and providing some experimental materials. We also thank Jean-Pierre Kinet for providing the transgenic mice expressing the human FcɛRIα. We are grateful to M. Lipscomb and J. Oliver from the UNM Asthma SCOR programs. This study was supported by NIH grant (AI-15251) to A.S. C.L.K. was supported by an American Lung Association-funded UNM Asthma Research Center grant, an Interest Section grant from the AAAAI and a Fellowship from the Parker B. Francis Foundation.

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  1. Daocheng Zhu and Christopher L. Kepley: D.Z. and C.L.K. contributed equally to this study.

Authors and Affiliations

  1. Division of Clinical Immunology/Allergy, Department of Medicine, The Hart and Louise Lyon Laboratory, University of California Los Angeles School of Medicine, Los Angeles, California, USA

    Daocheng Zhu, Min Zhang, Ke Zhang & Andrew Saxon

  2. Department of Pathology, University of New Mexico Health Sciences Center, Albuquerque, New Mexico, USA

    Christopher L. Kepley

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  1. Daocheng Zhu
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Correspondence to Andrew Saxon.

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Competing interests

A.S., D.Z. and K.Z. are listed as inventors on the patent for the use of bifunctional proteins to block allergic reactions. This patent is held by the University of California, their employer.

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Zhu, D., Kepley, C., Zhang, M. et al. A novel human immunoglobulin Fcγ–Fcɛ bifunctional fusion protein inhibits FcɛRI-mediated degranulation. Nat Med 8, 518–521 (2002). https://doi.org/10.1038/nm0502-518

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