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F(ab)′2-mediated neutralization of C3a and C5a anaphylatoxins: a novel effector function of immunoglobulins

Nature Medicine volume 9, pages 431–438 (2003)Cite this article

Abstract

High-dose intravenous immunoglobulin (IVIG) prevents immune damage by scavenging complement fragments C3b and C4b. We tested the hypothesis that exogenous immunoglobulin molecules also bind anaphylatoxins C3a and C5a, thereby neutralizing their pro-inflammatory effects. Single-cell calcium measurements in HMC-1 human mast cells showed that a rise in intracellular calcium caused by C3a and C5a was inhibited in a concentration-dependent manner by IVIG, F(ab)′2-IVIG and irrelevant human monoclonal antibody. C3a- and C5a-induced thromboxane (TXB2) generation and histamine release from HMC-1 cells and whole-blood basophils were also suppressed by exogenous immunoglobulins. In a mouse model of asthma, immunoglobulin treatment reduced cellular migration to the lung. Lethal C5a-mediated circulatory collapse in pigs was prevented by pretreatment with F(ab)′2-IVIG. Molecular modeling, surface plasmon resonance (SPR) and western blot analyses suggested a physical association between anaphylatoxins and the constant region of F(ab)′2. This binding could interfere with the role of C3a and C5a in inflammation.

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Figure 1: C3a- and C5a-induced [Ca2+]i responses in HMC-1 cells are inhibited by immunoglobulins.
Figure 2: Inhibition of C3a- and C5a-induced release of histamine and thromboxane from HMC-1 cells and whole-blood basophils.
Figure 3: Cellular infiltration of bronchoalveolar and lung parenchymal compartments of immunoglobulin-treated and C3-deficient mice.
Figure 4: Inhibition of C5a-induced lethal circulatory changes in pigs.
Figure 5: Physical association of anaphylatoxin and immunoglobulin molecules.

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Acknowledgements

The authors thank T. Semere for his help with HMC-1 cell cultures; M. Opperman for monoclonal antibodies against C3a and C5a; P. Gronski for purified F(ab)′2 immunoglobulin fragments and IVIG preparations (GammaVenin and Venimmun); R. Huber for the X-ray structures of the C3a molecule; E. Padlan for the X-ray and model structures of the Fab fragment of IgG; and B. Kelsall and P. Murphy for critical reading of the manuscript and helpful suggestions.

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Authors and Affiliations

  1. Neuronal Excitability Section, National Institute for Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA

    Milan Basta

  2. Section on Cellular Signaling, National Institute for Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA

    Fredric Van Goor & Stanko S. Stojilkovic

  3. Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Stefano Luccioli & Dean D. Metcalfe

  4. Computational Biophysics Section, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, USA

    Eric M. Billings

  5. Surgical Neurology Branch, National Institute for Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA

    Alexander O. Vortmeyer

  6. Department of Membrane Biochemistry, Walter Reed Army Institute of Research, Silver Spring, Maryland, USA

    Lajos Baranyi, Janos Szebeni & Carl R. Alving

  7. Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA

    Michael C. Carroll

  8. Laboratory of Immunoregulation, Office of Vaccines, Center for Biologics, Food and Drug Administration, Bethesda, Maryland, USA

    Ira Berkower

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Correspondence to Milan Basta.

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Basta, M., Van Goor, F., Luccioli, S. et al. F(ab)′2-mediated neutralization of C3a and C5a anaphylatoxins: a novel effector function of immunoglobulins. Nat Med 9, 431–438 (2003). https://doi.org/10.1038/nm836

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