Humanized Severe Combined Immunodeficient Mice as a Potential Model for the Study of Tolerance to Factor VIII

Summary

A Severe Combined Immunodeficient (SCID) mouse model has been established to evaluate experimental conditions leading to the production of factor VIII (FVIII) autoantibodies. To this end, we humanized 10 groups of 7 mice with peripheral blood mononuclear cells of 10 unrelated healthy blood donors (15 X 106 cells/mouse). Mice were injected with saline or immunized i.p. with 50 IU of a plasma derived human FVIII 24 h after reconstitution. Further immunization was made with 25 IU of FVIII every fortnight during 6 weeks and animals were sacrificed after 8 weeks. All reconstituted mice showed a spontaneous production of anti-FVIII antibodies in the absence of immunization with the corresponding antigen. However, no differences were observed regarding the quantity or the quality of these antibodies produced in the immunized or the saline group, indicating that tolerance to FVIII had been transferred with cell reconstitution. Affinity purified FVIII specific antibodies were capable of inhibiting FVIII activity and preventing the binding of FVIII to phospholipids in a dose-dependent manner. Immunoprecipitation experiments showed that the antibodies recognized only the C1 and C2 light chain domains. Since antibodies of interest can be found in the SCID mouse model and, moreover, since they are qualitatively comparable with the source donor’s antibodies, this model provides a tool to study the regulation of tolerance against self antigens in normal subjects and in acquired haemophilia patients.

Key words

SCID mice - factor VIII - tolerance

• References

• 1 Rossi F, Sultan Y, Kazatchkine M. Anti-idiotypes against autoantibodies and alloantibodies to VIII:C (anti-haemophilic factor) are present in therapeutic polyspecific normal immunoglobulins. Clin Exp Immunol 1988; 74: 311-6.
• 2 Gilles JGG, Arnout J, Vermylen J, Saint-Remy JMR. Anti-Factor VIII antibodies of haemophilic patients are frequently directed towards non-functional determinants and do not exhibit isotypic restriction. Blood 1993; 82: 2452-61.
• 3 Gilles JGG, Saint-Remy JMR. Healthy subjects produce both anti-Factor VIII and specific anti-idiotypic antibodies. J Clin Invest 1994; 94: 1496-505.
• 4 Algiman M, Dietrich G, Nydegger UE, Boeildieu D, Sultan Y, Kazatchkine MD. Natural antibodies to factor VIII (anti-hemophilic factor) in healthy individuals. Proc Natl Acad Sci USA 1992; 89: 3795-9.
• 5 Lundkvist I, Coutinho A, Varela F, Holmberg D. Evidence for a functional idiotypic network among natural antibodies in normal mice. Proc Natl Acad Sci USA 1989; 86: 5074-8.
• 6 Avrameas S, Guilbert B, Dighiero G. Natural antibodies against tubulin, actin, myoglobulin, thyroglobulin, fetuin, albumin and transferrin are present in normal human sera and monoclonal immunoglobulins from multiple myeloma and Waldenström’s macroglobulinemia may express similar antibody specificities. Ann Immunol (Inst Pasteur) 1981; 132C: 231-6.
• 7 Rossi F, Kazatchkine MD. Antiidiotypes against autoantibodies in pooled normal human polyspecific Ig. J Immunol 1989; 143: 4104-9.
• 8 Coutinho A, Kazatchkine MD, Avrameas S. Natural auto-antibodies. Curr Opin Immunol 1995; 07: 812-8.
• 9 Kerlero de Rosbo N, Hoffman M, Mendel I, Yust I, Kaye J, Bakimer R, Flechter S, Abramsky O, Milo R, Karni A, Ben-Nun A. Predominance of the autoimmune response to myelin oligodendrocyte glycoprotein (MOG) in multiple sclerosis: reactivity to the extracellular domain of MOG is directed against three main regions. Eur J Immunol 1997; 27: 3059-69.
• 10 Mosier DE, Gulizia RJ, Baird SM, Wilson DB. Transfer of a functional human immune system to mice with severe combined immunodeficiency. Nature 1988; 335: 256-9.
• 11 Jacquemin MG, Desqueper BG, Benhida A, Vander Elst L, Hoylaerts M, Bakkus M, Thielemans K, Arnout J, Peerlinck K, Gilles JGG, Vermylen J, Saint-Remy JMR. Mechanisms and kinetics of FVIII inactivation: study with an IgG4 monoclonal antibody derived from a hemophilia A patient with inhibitor. Blood 1998; 92: 496-506.
• 12 Jacquemin MG, Benhida A, Peerlinck K, Desqueper B, Vander Elst L, Lavend’homme R, d’Oiron R, Schwaab R, Bakkus M, Thielemans K, Gilles JG, Vermylen J, Saint-Remy JMR. A human antibody directed to the factor VIII (FVIII) C1 domain inhibits FVIII cofactor activity and binding to von Willebrand factor. Blood 2000; 95: 156-63.
• 13 Jacquemin MG, Peerlinck K, Gilles JG, Benhida A, Desqueper B, Lavend’homme R, Vermylen J, Saint-Remy JMR. Mutation Arg2150His in the factor VIII C1 domain alters the binding of factor VIII to von Willebrand factor and is responsible for a mild hemophilia A phenotype. Blood 1998; 92: 710a.
• 14 McCune JM, Namikawa R, Kaneshima H, Shultz LD, Lieberman M, Weissman IL. The SCID-hu Mouse: Murine model for the analysis of human hematolymphoid differentiation and function. Science 1988; 241: 1632-9.
• 15 Carlsson R, Martensson C, Kalliomäki S, Ohlin M, Borrebaeck CAK. Human peripheral blood lymphocytes transplanted into scid mice constitute an in vivo culture system exhibiting several parameters found in a normal humoral immune response and are a source of immunocytes for the production of human monoclonal antibodies. J Immunol 1992; 148: 1065-71.
• 16 Tary-Lehman M, Saxon A, Lehmann PV. The human immune system in hu-PBL-SCID mice. Immunol Today 1995; 16: 529-33.
• 17 Murray AG, Petzelbauer P, Hughes CCW, Costa J, Askenase P, Pober JS. Human T-cell-mediated destruction of allogeneic dermal microvessels in a severe combined immunodeficient mouse. Proc Natl Acad Sci USA 1994; 91: 9146-50.
• 18 Laulan A, Sauger A, Germain C, Montembault A, Sans I, Potentini-Esnault C, Chicane L, Gilard C, Sultan Y, Bartholeyns J. Frequency of human anti-FVIII antibodies in humanized SCID mice elicited by recombinant deleted factor VIII and by a plasma derived factor VIII. J Immunol Methods 1997; 210: 205-14.
• 19 Klinman NR. The “clonal selection hypothesis” and current concepts of B cell tolerance. Immunity 1996; 05: 189-95.
• 20 Nossal GJV. Negative selection of lymphocytes. Cell 1994; 76: 229-39.
• 21 Arai M, Scandella D, Hoyer LW. Molecular basis of factor VIII inhibition by human antibodies. Antibodies that bind to the factor VIII light chain prevent the interaction of factor VIII with phospholipid. J Clin Invest 1989; 83: 1978-84.
• 22 Prescott R, Nakai H, Saenko E, Scharrer I, Nilsson I, Humphries J, Hurst D, Bray G, Scandella D. the recombinate kogenate study groups. The inhibitor antibody response is more complex in hemophilia patients than in most nonhemophiliacs with factor VIII autoantibodies. Blood 1997; 89: 3663-71.
• 23 Shima M, Scandella D, Yoshioka A, Nakai H, Kamisue S, Terada S, Fukui H. A factor VIII neutralizing monoclonal antibody and a human inhibitor alloantibody recognizing epitopes in the C2 domain inhibit factor VIII binding to von Willebrand factor and to phosphatidylserine. Thromb Haemost 1993; 69: 240-6.
• 24 Saenko EL, Scandella D. A mechanism for inhibition of factor VIII binding to phospholipid by von Willebrand factor. J Biol Chem 1995; 270: 13826-33.
• 25 Scandella D, Gilbert GE, Shima M, Nakai H, Eagleson C, Felch M, Prescott R, Rajalakshmi KJ, Hoyer LW, Saenko E. Some factor VIII inhibitor antibodies recognize a common epitope corresponding to C2 domain amino acids 2248 through 2312, which overlap a phospholipid binding site. Blood 1995; 86: 1811-9.
• 26 Elder B, Lakich D, Gitschier J. Sequence of the murine factor VIII cDNA. Genomics 1993; 16: 374-9.
• 27 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.