Human Factor VIII Inhibitor Alloantibodies with a C2 Epitope Inhibit Factor Xa-catalyzed Factor VIII Activation: A new Anti-factor VIII Inhibitory Mechanism

Keiji Nogami; Midori Shima; Katsumi Nishiya; Yoshihiko Sakurai; Ichiro Tanaka; John C. Giddings; Evgueni L. Saenko; Akira Yoshioka

doi:10.1055/s-0037-1613026

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2002; 87(03): 459-465
DOI: 10.1055/s-0037-1613026

Review Article

Schattauer GmbH

Human Factor VIII Inhibitor Alloantibodies with a C2 Epitope Inhibit Factor Xa-catalyzed Factor VIII Activation: A new Anti-factor VIII Inhibitory Mechanism

Authors

Keiji Nogami

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
Midori Shima

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
Katsumi Nishiya

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
Yoshihiko Sakurai

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
Ichiro Tanaka

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
John C. Giddings

²Department of Haematology, University of Wales College of Medicine, Cardiff, U.K
Evgueni L. Saenko

³American Red Cross, Holland Laboratory, Rockville, M.D., USA
Akira Yoshioka

¹Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan

Abstract

Summary

Factor VIII (FVIII) inhibitor alloantibodies react with the A2, C2, or A3-C1 domains of FVIII and inactivate FVIII activity. We recently demonstrated that an anti-C2 monoclonal antibody with a Val²²⁴⁸Gly²²⁸⁵ epitope, inhibited factor Xa (FXa)-catalyzed FVIII activation, and that a FXa binding site for FVIII was located within residues Thr²²⁵³-Gln²²⁷⁰. In this study, we investigated whether anti-C2 alloantibodies inhibit FXa-catalyzed FVIII activation. Anti-C2 alloantibodies from four patients inhibited FVIII activation by FXa in onestage clotting assay. Furthermore, analysis by SDS-PAGE showed that all alloantibodies inhibited FVIII proteolytic cleavage by FXa independently of phospholipid. To confirm direct inhibition of FVIII and FXa interaction, we examined the effect of alloantibodies on FVIII binding to anhydro-FXa, a catalytically inactive FXa, in ELISA. All alloantibodies and C2-affinity purified F(ab)’₂ preparations inhibited FVIII binding to anhydro-FXa dose-dependently. Our results revealed a new inhibitory mechanism of FVIII, mediated by inhibition of FXa in the presence of anti-C2 alloantibodies.

Keywords

Factor VIII - factor Xa - hemophilia A - inhibitor - C2 domain

Full Text

References

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