Thromb Haemost 2006; 95(02): 341-347
DOI: 10.1160/TH05-08-0559
Animal Models
Schattauer GmbH

Tolerance to factor VIII in a transgenic mouse expressing human factor VIII cDNA carrying an Arg593 to Cys substitution

Wendy S. Bril*
1  Department of Plasma Proteins, Sanquin Research at CLB and Landsteiner Laboratory, AMC, University of Amsterdam, Amsterdam, The Netherlands
,
Pauline M. W. van Helden*
1  Department of Plasma Proteins, Sanquin Research at CLB and Landsteiner Laboratory, AMC, University of Amsterdam, Amsterdam, The Netherlands
,
Christina Hausl
2  BMT-Research, Vienna, Austria
,
Marleen G. Zuurveld
1  Department of Plasma Proteins, Sanquin Research at CLB and Landsteiner Laboratory, AMC, University of Amsterdam, Amsterdam, The Netherlands
,
Rafi U. Ahmad
3  Baxter BioScience, Vienna, Austria
,
Martine J. Hollestelle
1  Department of Plasma Proteins, Sanquin Research at CLB and Landsteiner Laboratory, AMC, University of Amsterdam, Amsterdam, The Netherlands
,
Pieter H. Reitsma
4  Department of Experimental Internal Medicine, AMC, Amsterdam, The Netherlands
,
Karin Fijnvandraat
5  Department of Pediatrics, EKZ/Children’s AMC, University of Amsterdam, The Netherlands
,
Rene A. W. van Lier
6  Department of Experimental Immunology, AMC, Amsterdam
,
Hans Peter Schwarz
3  Baxter BioScience, Vienna, Austria
,
Koen Mertens
1  Department of Plasma Proteins, Sanquin Research at CLB and Landsteiner Laboratory, AMC, University of Amsterdam, Amsterdam, The Netherlands
7  Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS) Utrecht, The Netherlands
,
Birgit M. Reipert
2  BMT-Research, Vienna, Austria
3  Baxter BioScience, Vienna, Austria
,
Jan Voorberg
1  Department of Plasma Proteins, Sanquin Research at CLB and Landsteiner Laboratory, AMC, University of Amsterdam, Amsterdam, The Netherlands
› Author Affiliations
Financial support: Supported by a grant from the Netherlands Organization of Science (NWO grant 902–26–204).
Further Information

Publication History

Received 16 August 2005

Accepted after resubmission 25 January 2005

Publication Date:
28 November 2017 (online)

Summary

Inhibitory antibodies develop in approximately 25% of patients with severe hemophilia A following treatment with factor VIII. In E-16KO or E-17KO mice, in which the factor VIII gene has been inactivated by insertion of a neo cassette, inhibitors develop following administration of factor VIII. Here, we describe the generation of transgenic mice expressing human factor VIII-R593C (huFVIII-R593C). Human factor VIII-R593C cDNA under control of a mouse albumin enhancer/promoter was injected into fertilized oocytes. Analysis of transgenic mice revealed that human factor VIII-R593C was expressed in the liver. Transgenic mice were crossed with factor VIII-deficient mice (E-16KO mice). In plasma of E-16KO mice antibodies were detected after five serial intravenous injections of factor VIII, while plasma of huFVIII-R593C/E-16KO mice did not contain detectable levels of antibodies. No antibody secreting cells were observed in either spleen or bone marrow of huFVIII-R593C/E-16KO mice. Also, factor VIII-specific memory B cells were not observed in the spleen of huFVIII-R593C/E-16KO mice. Analysis of T cell responses revealed that splenocytes derived of E-16KO mice secreted IL-10 and IFN-γ following restimulation with factor VIII in vitro. In contrast, no factor VIII-specific T cell responses were observed in huFVIII-R593C/E-16KO mice. These results indicate that huFVIII-R593C/E-16KO mice are tolerant to intravenously administered factor VIII. It is anticipated that this model may prove useful for studying immune responses in the context of factor VIII gene therapy.

* These authors contributed equally to this manuscript.