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Cleavage of anti-PF4/Heparin IgG by a bacterial protease and potential benefit in heparin-induced thrombocytopenia

Claire Kizlik-Masson, Quentin Deveuve, Yuhang Zhou, Caroline Vayne, Gilles Thibault, Steven E. McKenzie, Claire Pouplard, Stéphane Loyau, Yves Gruel and Jérôme Rollin

Key Points

  • Cleavage of the anti-PF4/heparin antibody hinge region by IdeS abolishes FcgammaRIIA-dependent cellular activation.

  • IdeS treatment prevents thrombus formation and thrombocytopenia induced by anti-PF4/heparin antibodies.

Abstract

Heparin-induced thrombocytopenia (HIT) is due IgG antibodies, which bind platelet factor 4 (PF4) modified by polyanions, such as heparin (H). IgG/PF4/polyanion complexes directly activate platelets via FcgammaRIIA receptors. A bacterial protease, IgG-degrading enzyme of Streptococcus pyogenes (IdeS), cleaves the hinge region of heavy chain IgG, abolishing its ability to bind FcgammaR, including FcgammaRIIA. We evaluated whether cleavage of anti-PF4/H IgG by IdeS could suppress the pathogenicity of HIT antibodies. IdeS quickly cleaved purified 5B9, a monoclonal chimeric anti-PF4/H IgG1, which led to the formation of single cleaved 5B9 (sc5B9), without any reduction in binding ability to the PF4/H complex. However, as compared with uncleaved 5B9, the affinity of sc5B9 for platelet FcgammaRIIA was greatly reduced, and sc5B9 was also unable to induce heparin-dependent platelet activation. In addition, incubating IdeS in whole blood containing 5B9 or HIT plasma samples led to cleavage of anti-PF4/H antibodies, which fully abolished the ability to induce heparin-dependent platelet aggregation and tissue factor mRNA synthesis by monocytes. Also, when whole blood was perfused in von Willebrand factor-coated microfluidic channels, platelet aggregation and fibrin formation induced by 5B9 with heparin was strongly reduced after IdeS treatment. Finally, IdeS prevented thrombocytopenia and hypercoagulability induced by 5B9 with heparin in transgenic mice expressing human PF4 and FcgammaRIIA receptors. In conclusion, cleavage of anti-PF4/H IgG by IdeS abolishes heparin-dependent cellular activation induced by HIT antibodies. IdeS injection could be a potential treatment for patients with severe HIT.

  • Submitted March 5, 2019.
  • Revision received March 15, 2019.
  • Accepted March 25, 2019.