Fine Mapping of Inhibitory Anti-factor V Antibodies Using Factor V C2 Domain Mutants

Tomonori Izumi; Suhng Wook Kim; Anne Greist; Sandra Macedo-Ribeiro; Pablo Fuentes-Prior; Wolfram Bode; William H. Kane; Thomas L. Ortel

doi:10.1055/s-0037-1615962

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2001; 85(06): 1048-1054
DOI: 10.1055/s-0037-1615962

Review Article

Schattauer GmbH

Fine Mapping of Inhibitory Anti-factor V Antibodies Using Factor V C2 Domain Mutants

Identification of Two Antigenic Epitopes Involved in Phospholipid Binding

Tomonori Izumi

¹Division of Hematology, Departments of Medicine and Pathology, Duke University Medical Center, Durham, North Carolina, USA

,

Suhng Wook Kim^*

¹Division of Hematology, Departments of Medicine and Pathology, Duke University Medical Center, Durham, North Carolina, USA

,

Anne Greist

²Indiana Hemophilia and Thrombosis Center, Indianapolis, Indiana, USA

,

Sandra Macedo-Ribeiro^†

³Max-Planck-Institut für Biochemie, Abteilung Strukturforschung, Martinsried, Germany

,

Pablo Fuentes-Prior

³Max-Planck-Institut für Biochemie, Abteilung Strukturforschung, Martinsried, Germany

,

Wolfram Bode

³Max-Planck-Institut für Biochemie, Abteilung Strukturforschung, Martinsried, Germany

,

William H. Kane

¹Division of Hematology, Departments of Medicine and Pathology, Duke University Medical Center, Durham, North Carolina, USA

,

Thomas L. Ortel

¹Division of Hematology, Departments of Medicine and Pathology, Duke University Medical Center, Durham, North Carolina, USA

› Author Affiliations

Abstract

Summary

Hemorrhagic factor V inhibitors frequently bind to the second C-type (C2) domain of factor V and interfere with phospholipid binding. To define specific residues recognized by inhibitors from four patients (one bovine thrombin-induced and three spontaneous antibodies), epitope mapping was performed using recombinant human factor V lacking most of the B-type domain (FV des B) and alanine-substituted mutants within the C2 domain (FV des B C2 mutants). FV des B C2 mutants located in the region between Lys²⁰⁶⁰ and Glu²⁰⁶⁹ were resistant to inhibition by three IgG preparations including the bovine thrombin-induced antibody in both prothrombinase and phospholipid-binding assays. In contrast, mutations at Lys²⁰⁸⁷ and Lys²⁰⁹²/ Glu²⁰⁹⁶ were significantly resistant to inhibition by the fourth IgG preparation in both prothrombinase and phospholipid-binding assays. These results confirm interference of phospholipid binding by hemorrhagic factor V inhibitors and support the role(s) of these residues in phospholipid binding.

Presented in part at the 41st Annual Meeting of the American Society of Hematology, New Orleans, Louisiana, December 3–7, 1999

Key words

Factor V - hemorrhagic inhibitor - epitope mapping - C2 domain - phospholipid binding

Full Text

References

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