The overall goal of these studies is to test the feasibility of, and lay the foundations for, specific immunosuppressive treatments of factor VIII (fVIII) antibody inhibitors. We propose to develop and test in hemophilia A mice (they have a disrupted fVIII gene) procedures for """"""""tolerization"""""""" of the CD4+ T helper cells involved in the synthesis of fVIII antibody inhibitors. Similar procedures were very effective in preventing and down regulating another Ab-mediated mouse syndrome, experimental myasthenia gravis. Also, we will investigate the epitope repertoire of human anti-fVIII CD4+ cells. If tolerization procedures will be effective in mice, knowledge of the CD4+ repertoire in humans will be needed for development of similar treatments of fVIII inhibitors for hemophilia A patients.
The specific aims will be: 1) To determine the epitope repertoire of anti-fVIII CD4+ cells in hemophilia A mice immunized with fVIII. We will challenge CD4+ spleen cells of fVIII immunized mice with overlapping synthetic peptides spanning the fVIII sequence. 2) To determine whether the anti-fVIII CD4+ cells in hemophilia A mice are Thl or Th2. These experiments will determine whether Thl or Th2 cells, or both, are necessary for synthesis of anti-fVIII Ab in general, and inhibitors in particular. This information will help the design of optimal tolerization procedure. 3) To use synthetic fVIII peptides comprising CD4+ epitope sequences, for tolerization procedures in hemophilia A mice. We will first test if nasal tolerization procedures will affect development of CD4+ and Ab response to fVIII. We will administer in the nasal cavities of the mice solutions of the synthetic fVIII CD4+ epitope peptides, prior to and during immunization with fVIII. If nasal administration will not be effective, we will attempt subcutaneous administration of the same peptides. 4) To investigate the epitope repertoire of human anti-fVIII CD4+ cells. We will challenge with the overlapping synthetic fVIII sequences used for the mice studies, blood CD4+ T cells from hemophilia A patients with and without antibody inhibitors, acquired autoimmune hemophilia patients and healthy subjects, which have ephemeral yet clearly detectable CD4+ responses to fVIII.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL061922-02
Application #
6056586
Study Section
Special Emphasis Panel (ZHL1-CSR-Q (S2))
Project Start
1998-09-30
Project End
2002-08-31
Budget Start
1999-09-29
Budget End
2000-08-31
Support Year
2
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Gharagozlou, Soheila; Sharifian, Ramazan A; Khoshnoodi, Jalal et al. (2009) Epitope specificity of anti-factor VIII antibodies from inhibitor positive acquired and congenital haemophilia A patients using synthetic peptides spanning A and C domains. Thromb Haemost 101:834-9
Hu, Genlin; Guo, Delan; Key, Nigel S et al. (2007) Cytokine production by CD4+ T cells specific for coagulation factor VIII in healthy subjects and haemophilia A patients. Thromb Haemost 97:788-94
Pratt, Kathleen P; Qian, Jiahua; Ellaban, Ekram et al. (2004) Immunodominant T-cell epitopes in the factor VIII C2 domain are located within an inhibitory antibody binding site. Thromb Haemost 92:522-8
Hu, G-L; Okita, D K; Conti-Fine, B M (2004) T cell recognition of the A2 domain of coagulation factor VIII in hemophilia patients and healthy subjects. J Thromb Haemost 2:1908-17
Reding, M T; Okita, D K; Diethelm-Okita, B M et al. (2004) Epitope repertoire of human CD4(+) T cells on the A3 domain of coagulation factor VIII. J Thromb Haemost 2:1385-94
Hu, G-L; Okita, D K; Diethelm-Okita, B M et al. (2003) Recognition of coagulation factor VIII by CD4+ T cells of healthy humans. J Thromb Haemost 1:2159-66
Reding, M T; Okita, D K; Diethelm-Okita, B M et al. (2003) Human CD4+ T-cell epitope repertoire on the C2 domain of coagulation factor VIII. J Thromb Haemost 1:1777-84
Reding, Mark T; Lei, Sijin; Lei, Howard et al. (2002) Distribution of Th1- and Th2-induced anti-factor VIII IgG subclasses in congenital and acquired hemophilia patients. Thromb Haemost 88:568-75
Reding, M T; Wu, H; Krampf, M et al. (2001) CD4+ T cells specific for factor VIII as a target for specific suppression of inhibitor production. Adv Exp Med Biol 489:119-34
Wu, H; Reding, M; Qian, J et al. (2001) Mechanism of the immune response to human factor VIII in murine hemophilia A. Thromb Haemost 85:125-33

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