PDF herunterladen
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
› InstitutsangabenFinancial support: Supported by a grant from the Netherlands Organization of Science (NWO grant 902–26–204).
Weitere Informationen

Publikationsverlauf

Received 16. August 2005

Accepted after resubmission 25. Januar 2005

Publikationsdatum:
28. November 2017 (online)

    Lizenzen und Reprints

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.

Keywords

Transgenic animals - haemophilia A/ B - factor VIII

* These authors contributed equally to this manuscript.


 

  • References

  • 1 Hoyer LW. Hemophilia A. N Engl J Med 1994; 330: 38-47.
    CrossrefPubMedGoogle Scholar
  • 2 Mannucci PM, Tuddenham EG. The hemophilias--from royal genes to gene therapy. N Engl J Med 2001; 344: 1773-9.
    CrossrefPubMedGoogle Scholar
  • 3 Lollar P. Pathogenic antibodies to coagulation factors. Part one: factor VIII and factor IX. J Thromb Haemost 2004; 02: 1082-95.
    CrossrefPubMedGoogle Scholar
  • 4 Bi L, Lawler AM, Antonarakis SE. et al. Targeted disruption of the mouse factor VIII gene produces a model of hemophilia A. Nat Genet 1995; 10: 119-21.
    CrossrefPubMedGoogle Scholar
  • 5 Bi L, Sarkar R, Naas T. et al. Further characterization of factor VIII-deficient mice created by gene targeting: RNA and protein studies. Blood 1996; 88: 3446-50.
    PubMedGoogle Scholar
  • 6 Muchitsch EM, Turecek PL, Zimmermann K. et al. Phenotypic expression of murine hemophilia. Thromb Haemost 1999; 82: 1371-3.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 7 Qian J, Borovok M, Bi L. et al. Inhibitor antibody development and T cell response to human factor VIII in murine hemophilia A. Thromb Haemost 1999; 81: 240-4.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 8 Reipert BM, Ahmad RU, Turecek PL. et al. Characterization of antibodies induced by human factor VIII in a murine knockout model of hemophilia A. Thromb Haemost 2000; 84: 826-32.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 9 Sasgary M, Ahmad RU, Schwarz HP. et al. Single cell analysis of factor VIII-specific T cells in hemophilic mice after treatment with human factor VIII. Thromb Haemost 2002; 87: 266-72.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 10 Hausl C, Maier E, Schwarz HP. et al. Long-term persistence of anti-factor VIII antibody-secreting cells in hemophilic mice after treatment with human factor VIII. Thromb Haemost 2002; 87: 840-5.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 11 Hausl C, Ahmad RU, Schwarz HP. et al. Preventing restimulation of memory B cells in hemophilia A: a potential new strategy for the treatment of antibody-dependent immune disorders. Blood 2004; 104: 115-22.
    CrossrefPubMedGoogle Scholar
  • 12 Qian J, Collins M, Sharpe AH. et al. Prevention and treatment of factor VIII inhibitors in murine hemophilia A. Blood 2000; 95: 1324-9.
    PubMedGoogle Scholar
  • 13 Rossi G, Sarkar J, Scandella D. Long-term induction of immune tolerance after blockade of CD40-CD40L interaction in a mouse model of hemophilia A. Blood 2001; 97: 2750-7.
    CrossrefPubMedGoogle Scholar
  • 14 Reipert BM, Sasgary M, Ahmad RU. et al. Blockade of CD40/CD40 ligand interactions prevents induction of factor VIII inhibitors in hemophilic mice but does not induce long lasting immune tolerance. Thromb Haemost 2001; 86: 1345-52.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 15 Qian J, Burkly LC, Smith EP. et al. Role of CD154 in the secondary immune response: the reduction of pre-existing splenic germinal centers and anti-factor VIII inhibitor titre. Eur J Immunol 2000; 30: 2548-54.
    CrossrefPubMedGoogle Scholar
  • 16 Lei TC, Scott DW. Induction of tolerance to factor VIII inhibitors by gene therapy with immunodominant A2 and C2 domains presented by B cells as Ig fusion proteins. Blood 2005; 105: 4865-70.
    CrossrefPubMedGoogle Scholar
  • 17 Bril WS, MacLean PE, Kaijen PH. et al. HLA class II genotype and factor VIII inhibitors in mild hemophilia A patients with an Arg593 to Cys mutation. Hemophilia 2004; 10: 509-14.
    CrossrefPubMedGoogle Scholar
  • 18 Fijnvandraat K, Turenhout EAM, Van den Brink EN. et al. The missense mutation Arg593→Cys is related to antibody formation in a patient with mild hemophilia A. Blood 1997; 89: 4371-7.
    PubMedGoogle Scholar
  • 19 Thompson AR, Murphy MEP, Liu M-L. et al. Loss of tolerance to endogenous factor VIII in a mild hemophilia A patient with an Arg593→Cys mutation. Blood 1997; 90: 1902-10.
    PubMedGoogle Scholar
  • 20 Roelse JC, de Laaf RT, Timmermans SM. et al. Intracellular accumulation of factor VIII induced by missense mutations Arg593→Cys and Asn618→Ser explains cross-reacting material-reduced hemophilia A. Br J Haematol 2000; 108: 241-6.
    CrossrefPubMedGoogle Scholar
  • 21 Needham M, Gooding C, Hudson K. et al. LCR/ MEL: a versatile system for high-level expression of heterologous proteins in erythroid cells. Nucleic Acids Res 1992; 20: 997-1003.
    CrossrefPubMedGoogle Scholar
  • 22 Pinkert CA, Ornitz DM, Brinster RL. et al. An albumin enhancer located 10 kb upstream functions along with its promoter to direct efficient, liver-specific expression in transgenic mice. Genes Dev 1987; 01: 268-76.
    CrossrefPubMedGoogle Scholar
  • 23 Wagenaar GT, van Vuuren AJ, Girma M. et al. Characterization of transgenic mice that secrete functional human proteinC inhibitor into the circulation. Thromb Haemost 2000; 83: 93-101.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 24 Hollestelle MJ, Thinnes T, Crain K. et al. Tissue distribution of factor VIII gene expression in vivo--a closer look. Thromb Haemost 2001; 86: 855-61.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 25 Leyte A, Mertens K, Distel B. et al. Inhibition of human coagulation factor VIII by monoclonal antibodies. Mapping of functional epitopes with the use of recombinant factor VIII fragments. Biochem J 1989; 263: 187-94.
    CrossrefPubMedGoogle Scholar
  • 26 Lenting PJ, Donath MJ, van Mourik JA. et al. Identification of a binding site for blood coagulation factor IXa on the light chain of human factor VIII. J Biol Chem 1994; 269: 7150-5.
    PubMedGoogle Scholar
  • 27 Kasper CK, Aledort LM, Counts RB. et al. A more uniform measurement of factor VIII inhibitors. Thromb Diath Haemorrh 1975; 34: 869-72.
    PubMedGoogle Scholar
  • 28 Billiau A, Matthys P. Modes of action of Freund’s adjuvants in experimental models of autoimmune diseases. J Leukoc Biol 2001; 70: 849-60.
    PubMedGoogle Scholar
  • 29 Evans GL, Morgan RA. Genetic induction of immune tolerance to human clotting factor VIII in a mouse model for hemophilia A. Proc Natl Acad Sci U S A 1998; 95: 5734-9.
    CrossrefPubMedGoogle Scholar
  • 30 Vigouroux S, Yvon E, Biagi E. et al. Antigen-induced regulatoryT cells. Blood 2004; 104: 26-33.
    CrossrefPubMedGoogle Scholar
  • 31 Jin DY, Zhang TP, Gui T. et al. Creation ofa mouse expressing defective human factor IX. Blood 2004; 104: 1733-9.
    CrossrefPubMedGoogle Scholar
  • 32 Sabatino DE, Armstrong E, Edmonson S. et al. Novel hemophilia B mouse models exhibiting a range of mutations in the Factor IX gene. Blood 2004; 104: 2767-2774.
    CrossrefPubMedGoogle Scholar
  • 33 Van den Brink EN, Timmermans SMH, Turenhout EAM. et al. Longitudinal analysis of factor VIII inhibitors in a previously untreated mild hemophilia A patient with an Arg593→Cys substitution. Thromb Haemost 1999; 81: 723-6.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 34 Rawle FE, Shi CX, Brown B. et al. Heterogeneity of the immune response to adenovirus-mediated factor VIII gene therapy in different inbred hemophilic mouse strains. J Gene Med 2004; 06: 1358-68.
    CrossrefPubMedGoogle Scholar
  • 35 Hay CR, Ollier W, Pepper L. et al. HLA class II profile:a weak determinant of factor VIII inhibitor development in severe haemophilia A. UKHCDO Inhibitor Working Party. Thromb Haemost 1997; 77: 234-7.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 36 Oldenburg J, Picard JK, Schwaab R. et al. HLA genotype of patients with severe haemophilia A due to intron 22 inversion with and without inhibitors of factor VIII. Thromb Haemost 1997; 77: 238-42.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 37 Jacquemin M, Vantomme V, Buhot C. et al. CD4+ T-cell clones specific for wild-type factor VIII: a molecular mechanism responsible for a higher incidence of inhibitor formation in mild/moderate hemophilia A. Blood 2003; 101: 1351-8.
    CrossrefPubMedGoogle Scholar
  • 38 Cook DN, Pisetsky DS, Schwartz DA. Toll-like receptors in the pathogenesis of human disease. Nat Immunol 2004; 05: 975-9.
    CrossrefPubMedGoogle Scholar
  • 39 Pipe SW, Saint-Remy JM, Walsh CE. New high technology products for the treatment of hemophilia. Hemophilia 2004; 10: 55-63.
    CrossrefPubMedGoogle Scholar
  • 40 Hough C, Lillicrap D. Gene therapy for hemophilia: an imperative to succeed. J Thromb Haemost 2005; 03: 1195-205.
    CrossrefPubMedGoogle Scholar