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Thromb Haemost 2002; 88(02): 221-229
DOI: 10.1055/s-0037-1613191
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Schattauer GmbH

Von Willebrand Factor Modulates Factor VIII Immunogenicity: Comparative Study of Different Factor VIII Concentrates in a Haemophilia A Mouse Model

Mathias Behrmann
1   Institute for Biophysical Chemistry and Biochemistry, Johann-Wolfgang-Goethe-University, Frankfurt am Main, Germany
,
John Pasi
2   Division of Haematology, University of Leicester, Leicester, UK
,
Jean-Marie R. Saint-Remy
3   Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium
,
Ronald Kotitschke
4   Biotest Pharma GmbH, Dreieich, Germany
,
Michael Kloft
4   Biotest Pharma GmbH, Dreieich, Germany
› Author Affiliations
Further Information

Publication History

Received 10 August 2001

Accepted after resubmission 30 April 2002

Publication Date:
07 December 2017 (online)

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Summary

The development of an immune response towards factor VIII (FVIII) remains the major complication of haemophilia A replacement therapy. Product-related risk factors have recently been identified on the basis of epidemiological studies, but the mechanism is not understood. To this end, various commercially available FVIII concentrates were administered by the IV route to FVIII-knockout mice and the resulting immune response was characterised. Significantly higher inhibitor titres (Bethesda assay) were observed for one recombinant FVIII and one plasma-derived FVIII product depleted in von Willebrand factor (VWF). Inhibitor titres were reduced upon pre-incubation of FVIII with purified VWF. Epitope specificity of anti-FVIII IgG was characterised using FVIII-fragments produced in E. coli. Concentrates with no or reduced VWF-level elicited antibodies recognising predominantly the acidic a1 and a3 regions. Addition of VWF prior to injection also modified the epitope specificity. FVIII concentrates, therefore, show qualitative and quantitative variations in immunogenicity, which are at least partly modulated by VWF.

Keywords

Haemophilia A - factor VIII - von Willebrand factor - animal model - immunogenicity
 

  • References

  • 1 Vehar GA, Keyt B, Eaton D, Rodriguez H, O’Brien DP, Rotblat F, Oppermann H, Keck R, Wood WI, Harkins RN, Tuddenham EGD, Lawn RM, Capon DJ. Structure of human factor VIII. Nature 1984; 312: 337-42.
    CrossrefPubMedGoogle Scholar
  • 2 Lenting PJ, van Mourik JA, Mertens K. The life cycle of coagulation factor VIII in view of its structure and function. Blood 1998; 92: 3983-96.
    PubMedGoogle Scholar
  • 3 Andersson LO, Forsman N, Huang K, Larsen K, Lundin A, Pavlu B, Sandberg H, Sewerin K, Smart J. Isolation and characterization of human factor VIII: molecular forms in commercial factor VIII concentrate, cryoprecipitate, and plasma. Proc Natl Acad Sci USA 1986; 83: 2979-83.
    CrossrefPubMedGoogle Scholar
  • 4 Hamer RJ, Koedam JA, Beeser-Visser NH, Bertina RM, van Mourik JA, Sixma JJ. Factor VIII binds to von Willebrand factor via its Mr-80.000 light chain. Eur J Biochem 1987; 166: 37-43.
    CrossrefPubMedGoogle Scholar
  • 5 Saenko EL, Shima M, Rajalakshmi KJ, Scandella D. A role for the C2 domain of factor VIII in binding to von Willebrand factor. J Biol Chem 1994; 269: 11601-5.
    PubMedGoogle Scholar
  • 6 Koedam JA, Meijers JC, Sixma JJ, Bouma BN. Inactivation of human factor VIII by activated protein C: Cofactor activity of protein S and protective effect of von Willebrand factor. J Clin Invest 1988; 82: 1236-43.
    CrossrefPubMedGoogle Scholar
  • 7 Koedam JA, Hamer RJ, Beeser-Visser NH, Bouma BN, Sixma JJ. The effect of von Willebrand factor on activation of factor VIII by factor Xa. Eur J Biochem 1990; 189: 229-34.
    CrossrefPubMedGoogle Scholar
  • 8 Scharrer I, Bray GL, Neutzling O. Incidence of inhibitors in haemophilia A patients a review of recent studies of recombinant and plasma-derived factor VIII concentrates. Hemophilia 1999; 05: 145-54.
    CrossrefPubMedGoogle Scholar
  • 9 Fulcher C, De Graaf SMahoney, Zimmermann T. Factor VIII inhibitor IgG subclass and fVIII polypeptide specificity determined by immunoblotting. Blood 1987; 69: 1475-80.
    PubMedGoogle Scholar
  • 10 Gilles JG, Arnout J, Vermylen J, Saint-Remy JM. Anti-factor VIII antibodies of hemophiliac patients are frequently directed towards nonfunctional determinants and do not exhibit isotypic restriction. Blood 1993; 82: 2452-61.
    PubMedGoogle Scholar
  • 11 Lollar P, Parker ET, Curtis JE, Helgerson SL, Hoyer LW, Scott ME, Scandella D. Inhibition of human factor VIIIa by anti-A2 subunit antibodies. J Clin Invest 1994; 93: 2497-504.
    CrossrefPubMedGoogle Scholar
  • 12 Scandella D, DeGraaf SMahoney, Mattingly M, Roeder D, Timmons L, Fulcher CA. Epitope mapping of human factor VIII inhibitor antibodies by deletion analysis of factor VIII fragments expressed in Escherichia coli . Proc Natl Acad Sci USA 1988; 85: 6152-6.
    CrossrefPubMedGoogle Scholar
  • 13 Foster PA, Fulcher CA, Houghten RA, Zimmerman TS. An immunogenic region within residues Val1670-Glu1684 of the factor VIII light chain induces antibodies which inhibit binding of factor VIII to von Willebrand factor. J Biol Chem 1988; 263: 5230-4.
    PubMedGoogle Scholar
  • 14 Brackmann H-H, Schwaab R, Effenberger W, Hess L, Hanfland P, Oldenburg J. Antibodies to factor VIII in Hemophilia A patients. Vox Sang 2000; 78 (02) 187-90.
    PubMedGoogle Scholar
  • 15 Rosendaal FR, Nieuwenhuis HK, van den Berg HM, Heijboer H, Mauser EPBunschoten, van der Meer J, Smit C, Strengers PFW, Briet E. and the Dutch Hemophilia Study Group. A sudden increase in factor VIII inhibitor development in multitransfused hemophilia A patients in the Netherlands. Blood 1993; 81: 2180-6.
    PubMedGoogle Scholar
  • 16 Peerlinck K, Arnout J, Gilles JG, Saint-Remy JM, Vermylen J. A higher than expected incidence of factor VIII inhibitors in multitransfused Haemophilia A patients treated with an intermediate purity pasteurized factor VIII concentrate. Thromb Haemost 1993; 69: 115-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Laub R, Di Giambattista M, Fondu P, Brackmann H-H, Lenk H, Saenko EL, Felch M, Scandella D. Inhibitors in German haemophilia A patients treated with a double virus inactivated factor VIII concentrate bind the C2 domain of FVIII light chain. Thromb Haemost 1999; 81: 39-44.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Bi L, Lawler AM, Antonarakis SE, High KA, Gearhart JD, Kazazian HH. Targeted disruption of the mouse factor VIII gene produces a model of haemophilia A. Nat Genet 1995; 10: 119-21.
    CrossrefPubMedGoogle Scholar
  • 19 O’Brien DP, Johnson D, Byfield P, Tuddenham EGD. Inactivation of factor VIII by factor IXa. Biochemistry 1992; 31: 2805-12.
    CrossrefPubMedGoogle Scholar
  • 20 Kasper CK, Aledort LM, Counts RB, Edson JR, Fratantoni J, Green D, Hampton JW, Hilgartner MW, Lazerson J, Levine PH, McMillan CW, Pool JG, Shapiro SS, Shulman NR, van Eys J. A more uniform measurement of factor VIII inhibitors. Thromb Diath Haemorrh 1975; 34: 869-72.
    PubMedGoogle Scholar
  • 21 Jacquemin MG, Desqueper BG, Benhida A, Van der Elst L, Hoylaerts MF, Bakkus M, Thielemans K, Arnout J, Peerlinck K, Gilles JGG, Vermylen J, Saint-Remy JM. Mechanism and kinetics of factor VIII inactivation: study with an IgG4 monoclonal antibody derived from a hemophilia A patient with inhibitor. Blood 1998; 92: 496-506.
    PubMedGoogle Scholar
  • 22 Grieco F, Hay JM, Hull R. An improved procedure for the purification of protein fused with glutathione S-transferase. Biotechniques 1992; 13: 856-8.
    PubMedGoogle Scholar
  • 23 Lawrence J. Recombinate®; viral safety and final product manufacturing testing and specifications. Ann Hematol 1994; 68: S21-4.
    CrossrefPubMedGoogle Scholar
  • 24 Jaquemin M, Lavend’homme R, Benhida A, Vanzieleghem B, d´Oiron R, Lavergne J-M, Brackmann HH, Schwaab R, VandenDriesche T, Chuah MKL, Hoylaerts M, Gilles JGG, Peerlinck K, Vermylen J, Saint-Remy JM. A novel cause of mild/moderate hemophilia A: mutations scattered in the factor VIII C1 domain reduce factor VIII binding to von Willebrand factor. Blood 2000; 96: 958-65.
    PubMedGoogle Scholar
  • 25 Singh I, Smith A, Vanzieleghem B, Collen D, Burnand K, Saint-Remy JM, Jaquemin M. Antithrombotic effects of controlled inhibition of factor VIII with a partially inhibitory human monoclonal antibody in a murine vena cava thrombosis model. Blood 2002; 99: 3235-40.
    CrossrefPubMedGoogle Scholar
  • 26 Bi L, Sarkar R, Naas T, Lawler AM, Pain J, Shumaker SL, Bedian V, Kazazian HH. Further characterization of factor VIII-deficient mice created by gene targeting: RNA and protein studies. Blood 1996; 88: 3446-50.
    PubMedGoogle Scholar
  • 27 Reipert BM, Ahmad RU, Turecek PL, Schwarz HP. Characterization of antibodies induced by human factor VIII in a murine kockout model of hemophilia A. Thromb Haemost 2000; 84: 826-32.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Wu H, Reding M, Qian J, Okita DK, Parker E, Lollar P, Hoyer LW, Conti-Fine BM. Mechanism of the immune response to human factor VIII in murine hemophilia A. Thromb Haemost 2001; 85: 125-33.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Kreuz W, Escuriola-Ettingshausen C, Martinez-Saguer I, Kaiml M, Kornhuber B. Epidemiology of inhibitor development in haemophilia a patients treated with virus-inactivated plasma-derived clotting factor concentrates. Vox Sang 1999; 77 (01) 3-8.
    CrossrefPubMedGoogle Scholar
  • 30 Qian J, Borovok M, Bi L, Kazazian HH, Hoyer LW. Inhibitor antibody development and T cell response to human factor VIII in murine Hemophilia A. Thromb Haemost 1999; 81: 240-4.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 31 Josic D, Buchacher A, Kannicht C, Lim YP, Loster K, Pock K, Robinson S, Schwinn H, Stadler M. Degradation products of factor VIII which can lead to increased immunogenicity. Vox Sang 1999; 77 (01) 90-9.
    CrossrefPubMedGoogle Scholar
  • 32 Hironaka T, Furukawa K, Esmon PC, Fournel MA, Sawada S, Kato M, Minaga T, Kobata A. Comparative study of the sugar chains of factor VIII purified from human plasma and from the culture media of recombinant baby hamster kidney cells. J Biol Chem 1992; 267: 8012-20.
    PubMedGoogle Scholar
  • 33 Hodge G, Flower R, Han P. Effect of factor VIII concentrate on leucocyte cytokine production: characterization of TGF-β as an immunomodulatory component in plasma-derived factor VIII concentrate. Br J Haematol 1999; 106: 784-91.
    CrossrefPubMedGoogle Scholar
  • 34 Suzuki T, Arai M, Armano K, Kagawa K, Fukutake K. Factor VIII inhibitor antibodies with C2 domain specificity are less inhibitory to factor VIII complexed with von Willebrand factor. Thromb Haemost 1996; 76: 749-54.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Gensana M, Altisent C, Aznar JA, Casana P, Hernandez F, Jorquera JI, Magallon M, Massot M, Puig L. Influence of von Willebrand factor on the reactivity of human factor VIII inhibitors with factor VIII. Haemophilia 2001; 07: 369-74.
    CrossrefPubMedGoogle Scholar
  • 36 Gilles JG, Lavend’homme R, Peerlinck K, Jacquemin MG, Hoylaerts M, Jorieux S, Mazurier C, Vermylen J, Saint-Remy J-MR. Some factor VIII (fVIII) inhibitors recognise a fVIII epitope(s) that is present only on fVIIIVWF complexes. Thromb Haemost 1999; 82: 40-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 37 Kawasaki T, Kaida T, Arnout J, Vermylen J, Hoylaerts MF. A new animal model of thrombophilia confirms that high plasma factor VIII levels are thrombogenic. Thromb Haemost 1999; 81: 306-11.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 38 Saenko EL, Scandella D. The acidic region of the factor VIII light chain and the C2 domain together form the high affinity binding site for von Willebrand factor. J Biol Chem 1997; 272: 18007-14.
    CrossrefPubMedGoogle Scholar
  • 39 Prescott R, Nakai H, Saenko EL, Scharrer I, Nilsson IM, Humphries JE, Hurst D, Bray G, Scandella D. and the Recombinate and Kogenate Study Groups. The inhibitor antibody response is more complex in hemophilia A patients than in most nonhemophiliacs with factor VIII autoantibodies. Blood 1997; 89: 3663-71.
    PubMedGoogle Scholar
  • 40 Vanzieleghem B, Gilles JG, Desqueper B, Vermylen J, Saint-Remy J-M. Humanized severe combined immunodeficient mice as a potential model for the study of tolerance to factor VIII. Thromb Haemost 2000; 83: 833-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 41 Vlot AJ, Koppelman SJ, Meijers JCM, Damas C, van den Berg HM, Bouma BN, Sixma JJ, Willems GM. Kinetics of factor VIII- von Willebrand factor association. Blood 1996; 87: 1809-16.
    PubMedGoogle Scholar
  • 42 Raut S, Di Gambattista M, Bevan SA, Hubbard AR, Barrowcliffe TW, Laub R. Modification of factor VIII in therapeutic concentrates after virus inactivation by solvent-detergent and pasteurisation. Thromb Haemost 1998; 80: 624-31.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 43 Saenko E, Josic D, Stadler M, Sarafanov A, Lim Y-P, Shima M, Ananyeva N, Schwinn H. Molecular modifications in factor VIII concentrates produced from different plasma pools. Thromb Res 2001; 101: 501-11.
    CrossrefPubMedGoogle Scholar
  • 44 Raut S, Weller L, Barrowcliffe TW. Phospholipid binding of factor VIII in different therapeutic concentrates. Br J Hematol 1999; 107: 323-9.
    CrossrefPubMedGoogle Scholar