Thromb Haemost 2001; 85(01): 125-133
DOI: 10.1055/s-0037-1612915
Review Article
Schattauer GmbH

Mechanism of the Immune Response to Human Factor VIII in Murine Hemophilia A

Huiyun Wu
1  Department of Biochemistry, Molecular Biology and Biophysics, and Department of Medicine, University of Minnesota, Minneapolis-St. Paul, MN, GA, USA
,
Mark Reding
1  Department of Biochemistry, Molecular Biology and Biophysics, and Department of Medicine, University of Minnesota, Minneapolis-St. Paul, MN, GA, USA
2  Department of Medicine, University of Minnesota, Minneapolis-St. Paul, MN, GA, USA
,
Jiahua Qian
1  Department of Biochemistry, Molecular Biology and Biophysics, and Department of Medicine, University of Minnesota, Minneapolis-St. Paul, MN, GA, USA
,
David K. Okita
1  Department of Biochemistry, Molecular Biology and Biophysics, and Department of Medicine, University of Minnesota, Minneapolis-St. Paul, MN, GA, USA
,
Ernie Parker
4  Division of Hematology-Oncology, Department of Medicine, Emory University, Atlanta, GA, USA
,
Pete Lollar
4  Division of Hematology-Oncology, Department of Medicine, Emory University, Atlanta, GA, USA
,
Leon W. Hoyer
3  Holland Laboratory, American Red Cross, Rockville, MD, GA, USA
,
Bianca M. Conti-Fine5
1  Department of Biochemistry, Molecular Biology and Biophysics, and Department of Medicine, University of Minnesota, Minneapolis-St. Paul, MN, GA, USA
› Author Affiliations
Supported by the NHLBI grants HL61922 (to B.M.C.-F.), and HL46215 (to P.L.). M.T.R. is the recipient of a Judith Graham Pool Postdoctoral Research Fellowship from the National Hemophilia Foundation.
Further Information

Publication History

Received 07 March 2000

Accepted after resubmission 08 August 2000

Publication Date:
08 December 2017 (online)

Summary

Mice genetically deficient in factor VIII (fVIII) are a model of hemophilia A. As a first step to reproduce in this mouse model what occurs over time in hemophilia A patients treated with human fVIII (hfVIII), we have investigated the time course and the characteristics of their immune response to hfVIII, after multiple intravenous injections. Anti-hfVIII antibodies appeared after four to five injections. They were IgG1 and to a lesser extent IgG2, indicating that they were induced by both Th2 and Th1 cells. Inhibitors appeared after six injections. CD4+ enriched splenocytes from hfVIII-treated mice proliferated in response to fVIII and secreted IL-10: in a few mice they secreted also IFN-γ and in one mouse IL-4, but never IL-2. A hfVIII-specific T cell line derived from hfVIII-treated mice secreted both IL-4 and IFN-γ, suggesting that it included both Th1 and Th2 cells. CD4+ enriched splenocytes of hfVIII-treated mice recognized all hfVIII domains. Thus, hemophilic mice develop an immune response to hfVIII administered intravenously similar to that of hemophilia A patients. Their anti-hfVIII antibodies can be inhibitors and belong to IgG subclasses homologous to those of inhibitors in hemophilic patients; their anti-hfVIII CD4+ cells recognize a complex repertoire and both Th1 and Th2 cytokines, and especially IL-10, may drive the antibody synthesis.

Abbreviations used: antibodies, Ab; antigen presenting cells, APC; Arbitrary Units, AU; enzyme-linked immunosorbant assay, ELISA; factor VIII, fVIII; human factor VIII, hf VIII; intravenous, i.v.; optical density, OD; polymerase chain reaction, PCR; phosphate buffered saline solution, PBS; PBS containing 3% bovine serum albumin, PBS/BSA; PBS containing 0.05% polyoxyethylene sorbitan monolaurate, PBS/Tween-20; phytohemoagglutinin, PHA; stimulation index, SI

5 Previously known as Bianca M. Conti-Tronconi