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DOI: 10.1055/s-0037-1613094
Long-term Persistence of Anti-factor VIII Antibody-secreting Cells in Hemophilic Mice after Treatment with Human Factor VIII
Publication History
Received
19 November 2001
Accepted
28 January 2002
Publication Date:
11 December 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/200205/lookinside/thumbnails/10-1055-s-0037-1613094-1.jpg)
Summary
The analysis of anti-factor VIII (FVIII) antibody-secreting cells (ASC) at different anatomic sites provides valuable information about the nature of the anti-FVIII immune response in hemophilic mice after treatment with human FVIII. An Elispot system is described that is suitable for analyzing frequencies and IgG subclasses of anti-FVIII ASC at the single-cell level. Hemophilic mice were treated with four doses of FVIII. Anti-FVIII antibodies in blood as well as anti-FVIII ASC in spleen and bone marrow were analyzed after each dose of FVIII and subsequently up to 22 weeks after termination of the FVIII treatment. Anti-FVIII ASC first appeared in the spleen where they were detectable after two intravenous doses of FVIII. Their appearance correlated with that of anti-FVIII antibodies in blood plasma. Anti- FVIII ASC in bone marrow were detectable after three doses of FVIII and were probably cells that initially formed in the spleen and subsequently migrated to the bone marrow. Whereas the frequency of anti- FVIII ASC in the spleen increased up to the fourth dose of FVIII and declined thereafter, in the bone marrow it remained constant for up to at least 22 weeks after the termination of the FVIII treatment. Titers of anti-FVIII antibodies in blood plasma increased up to the fourth dose of FVIII, then remained high constantly for 14 weeks and decreased but the antibodies were still detectable for up to at least 22 weeks after the fourth dose of FVIII. The IgG-subclass distribution of anti-FVIII ASC was similar in spleen and bone marrow and matched the subclasses of anti-FVIII antibodies in blood plasma indicating that both organs contribute to circulating antibodies in the blood.
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References
- 1 Hoyer LW. Hemophilia A. N Engl J Med 1994; 330: 39-47.
- 2 Hoyer LW. The incidence of factor VIII inhibitors in patients with severe hemophilia A. In: Aledort LM, Hoyer LW, Lisher JM, Reisner HM, White GC. eds. Inhibitors to coagulation factors. New York: Plenum; 1995: 35-45.
- 3 Ehrenforth S, Kreuz W, Scharrer I, Linde R, Funk M, Güngör T, Krackhardt B, Kornhuber B. Incidence of development of factor VIII and factor IX inhibitors in haemophiliacs. Lancet 1992; 339: 594-8.
- 4 Schwaab R, Brackmann HH, Meyer C, Seehafer J, Kirchgesser M, Haack A, Olek K, Tuddenham EG, Oldenburg J. Haemophilia A: Mutation type determines risk of inhibitor formation. Thromb Haemost 1995; 74: 1402-6.
- 5 Tuddenham EG, McVey JH. The genetic basis of inhibitor development in hemophilia A. Haemophilia 1998; 04: 543-5.
- 6 Fakharzadeh SS, Kazazian HH. Correlation between factor VIII genotype and inhibitor development in hemophilia A. Sem Thromb Hemost 2000; 26: 167-71.
- 7 Bi L, Lawler AM, Antonarakis SE, High KA, Gearhart JD, Kazazian Jr HH. Targeted disruption of the mouse factor VIII gene produces a model of haemophilia A. Nat Genet 1995; 10: 119-21.
- 8 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.
- 9 Qian J, Borovok M, Bi L, Kazazian Jr HH, Hoyer LW. Inhibitor development and T cell response to human factor VIII in murine haemophilia A. Thromb Haemost 1999; 81: 240-4.
- 10 Reipert BM, Ahmad RU, Turecek PL, Schwarz HP. FVIII neutralizing antibodies in murine hemophilia A as a model for immunotherapeutic intervention studies. FASEB J 2000; 14: A1130.
- 11 Reipert BM, Ahmad RU, Turecek PL, Schwarz HP. Characterization of antibodies induced by human factor VIII in a murine knockout model of hemophilia A. Thromb Haemost 2000; 84: 826-32.
- 12 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.
- 13 Manz RA, Thiel A, Radbruch A. Lifetime of plasma cells in the bone marrow. Nature 1997; 388: 133-4.
- 14 Slifka MK, Antia R, Withmire JK, Ahmed R. Humoral immunity due to long-lived plasma cells. Immunity 1998; 08: 363-72.
- 15 Manz RA, Löhning M, Cassese G, Thiel A, Radbruch A. Survival of longlived plasma cells is independent of antigen. Intern Immunol 1998; 11: 1703-11.
- 16 Muchitsch EM, Turecek PL, Zimmermann K, Pichler L, Auer W, Richter G, Gritsch H, Schwarz HP. Phenotypic expression of murine hemophilia (letter). Thromb Haemost 1999; 82: 1371-3.
- 17 Slifka MK, Ahmed R. Limiting dilution analysis of virus-specific memory B cells by an Elispot assay. J Immunol Methods 1996; 199: 37-46.
- 18 Möller SA, Borrebaeck CAK. A filter immuno-plaque assay for the detection of antibody-secreting cells in vitro. J Immunol Methods 1985; 79: 195-204.
- 19 Slifka MK, Matloubian M, Ahmed R. Bone marrow is the major site of long-term antibody production after acute viral infection. J Virol 1995; 69: 1895-902.
- 20 Talbot PJ, Buchmeier MJ. Catabolism of homologous murine monoclonal hybridoma IgG antibodies in mice. Immunology 1987; 60: 485-9.
- 21 Vieira P, Rajewsky K. The half-lives of serum immunoglobulins in adult mice. Eur J Immunol 1988; 18: 313-6.
- 22 Ochsenbein AF, Pinschewer DD, Sierro S, Horvath E, Hengartner H, Zinkernagel RM. Protective long-term antibody memory by antigen-driven T help-dependent differentiation of long-lived memory B-cells to shortlived plasma cells independent of secondary lymphoid organs. PNAS 2000; 97: 13263-8.
- 23 Gray D, Siepmann K, van Eden D, Poudrier J, Wykes M, Jainandunsing S, Bergthorsdottir S, Dullforce P. B-T lymphocyte interaction in the generation and survival of memory cells. Immunol Rev 1996; 150: 45-61.