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

Keiji Nogami
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Midori Shima
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Katsumi Nishiya
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Yoshihiko Sakurai
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Ichiro Tanaka
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
John C. Giddings
2   Department of Haematology, University of Wales College of Medicine, Cardiff, U.K
,
Evgueni L. Saenko
3   American Red Cross, Holland Laboratory, Rockville, M.D., USA
,
Akira Yoshioka
1   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
› Author Affiliations
Further Information

Publication History

Received 17 August 2001

Accepted after revission 14 November 2001

Publication Date:
14 December 2017 (online)

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.

 
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