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Cell surface expression of a human IgC Fc chimera activates macrophages through Fc receptors

Nature Biotechnology volume 16, pages 1357–1360 (1998)Cite this article

Abstract

Antibody-dependent cell-mediated cytotoxicity plays an important role in the macrophage-mediated destruction of target cells. While the selectivity is based on antibody specificity, the lytic attack is triggered by Fc receptor–mediated respiratory burst. To mimic IgG opsonization, a chimeric antibody-like molecule, containing human IgG1 Fc, was expressed on the surface of mammalian cells. The transmembrane domain of the human transferrin receptor was fused in-frame to the N-terminus of the second and third domains of human IgG1 heavy-chain constant region. This fusion molecule was designed to take advantage of the type II membrane anchor property of the transferrin receptor to express the Fc portion of the molecule in a reverse orientation, such that the Fc portion projected away from the cell surface. This is in contrast to the conventional cell surface IgG, which is anchored by a C-terminal type I transmembrane domain. The cell surface expressed reverse Fc no longer activated complement, but retained Fc receptor–binding capability and activated superoxide production by macrophages. This activity was completely blocked by an FcγR I–specific monoclonal antibody.

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Figure 1: Design rationale.
Figure 2: Expression of reverse hTR–FcΔH in BHK cells.
Figure 3: Effect of cell surface IgG Fc on superoxide production.
Figure 4: Effect of anti-FcγRI monoclonal antibody F(ab´)2 on BHK-FcΔH induced superoxide production.
Figure 5: Effect of BHK-FcΔH in complement activation.

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Acknowledgements

The authors thank William F. Hickey, James R. Dasch, Paul A. Johnston, Edward E. Baetge, and Joseph P. Hammang for helpful discussions, and Paula M. Krueger and Jennifer Jackson for their technical assistance. The authors also thank Mark D. Lindner for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Immunology, CytoTherapeutics, Inc., Lincoln, 02865, RI

    Paul F. Stabila, Shou C. Wong, Faith A. Kaplan & Weng Tao

  2. Department of Cell and Molecular Biology, CytoTherapeutics Inc., Lincoln, 02865, RI

    Shou C. Wong

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  1. Paul F. Stabila
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Correspondence to Weng Tao.

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Stabila, P., Wong, S., Kaplan, F. et al. Cell surface expression of a human IgC Fc chimera activates macrophages through Fc receptors. Nat Biotechnol 16, 1357–1360 (1998). https://doi.org/10.1038/4339

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