Recombinant factor VIII and factor VIII-von Willebrand factor complex do not present danger signals for human dendritic cells

Summary

Several lines of evidence have shown that antibody responses to coagulation factor VIII (FVIII) in patients with hemophilia A depend on the help of activated CD4⁺ T cells. The primary activation of CD4⁺ T cells requires interaction with mature dendritic cells (DCs) that present antigenic peptides in the context of MHC class II and express costimulatory molecules. Maturation of DCs requires danger signals provided by exogenous or endogenous stimuli such as pathogen-derived products or inflammatory cytokines. We asked the question whether FVIII itself, FVIII complexed with von Willebrand factor (VWF) or thrombin-activated FVIII contain danger signals for human DCs that induce the upregulation of costimulatory molecules or the expression of proinflammatory cytokines necessary for effec tive activation of CD4⁺ T cells. Human peripheral monocytes were differentiated into DCs. FVIII, thrombin-activated FVIII, VWF, VWF-FVIII, lipopolysaccharide (LPS), LPS+FVIII, LPS+VWF or LPS+FVIII-VWF were added either on day 0 or on day 5 of differentiation cultures. Differentiation markers, cytokines in cell culture supernatants and the capacity of DCs to stimulate autologous and allogeneic T cells were analysed after seven days of differentiation cultures. Our results indicate that neither FVIII, thrombin-activated FVIII, VWF nor a complex of FVIII and VWF modulate the maturation of human DCs or their capacity to stimulate autologous or allogeneic T cells. We conclude that neither of these proteins present danger signals to human DCs.

Keywords

Hemophilia A - FVIII inhibitors - human dendritic cells - danger signals - factor VIII

• References

• 1 Hoyer LW. Hemophilia A. New Engl J Med 1994; 330: 39-47.
• 2 Hoyer LW. The incidence of factor VIII inhibitors in patients with severe hemophilia A. In: Inhibitors to coagulation factors. New York: Plenum; 1995: 35-45.
• 3 Ehrenforth S, Kreuz W, Scharrer I. et al. Incidence of development of factor VIII and factor IX inhibitors in haemophiliacs. Lancet 1992; 339: 594-8.
• 4 Lollar P. Pathogenic antibodies to coagulation factors. Part one: Factor VIII and Factor IX. J Thromb Haemost 2004; 02: 1082-95.
• 5 Banchereau J, Steinman RM. Dendritic cells and the control of immunity. Nature 1998; 392: 245-52.
• 6 Steinman RM. The control of immunity and tolerance by dendritic cells. Pathol Biol 2003; 51: 59-60.
• 7 Lipscomb M, Masten BJ. Dendritic cells: immune regulators in health and disease. Physiol Rev 2002; 82: 97-130.
• 8 Dresser DW. Specific inhibition of antibody production. II. Paralysis induced in adult mice by small quantities of protein antigen. Immunology 1962; 05: 131-8.
• 9 Schlokat U, Fischer BE, Herlitschka S. et al. Production of highly homogeneous and structurally intact recombinant von Willebrand factor multimers by furinmediated propeptide removal in vitro . Biotechnol Appl Biochem 1996; 24: 257-67.
• 10 Plaimauer B, Schlokat U, Turecek PL. et al. Recombinant von Willebrand Factor: preclinical development. Semin Thromb Hemost 2001; 27: 395-403.
• 11 Turecek PL, Pichler L, Auer W. et al. Evidence for extracellular processing of pro-von Willebrand factor after infusion in animals with and without severe von Willebrand disease. Blood 1999; 94: 1637-47.
• 12 Schwarz HP, Schlokat U, Mitterer A. et al. Recombinant von Willebrand factor-insight into structure and function through infusion studies in animals with severe von Willebrand disease. Semin Thromb Hemost 2002; 28: 215-26.
• 13 Pickl WF, Majdic O, Kohl P. et al. Molecular and functional characteristics of dendritic cells generated from highly purified CD14+ peripheral blood monocytes. J Immunol 1996; 157: 3850-9.
• 14 Visintin A, Mazzoni A, Spitzer JH. et al. Regulation of Toll-like receptors in human monocytes and dendritic cells. J Immunol 2001; 166: 249-55.
• 15 Kaufman RJ, Wasley LC, Dorner AJ. Synthesis, processing, and secretion of recombinant human factor VIII expressed in mammalian cells. J Biol Chem 1988; 263: 6352-62.
• 16 Hill-Eubanks DC, Parker CG, Lollar P. Differential proteolytic activation of factor VIII-von Willebrand factor complex by thrombin. Proc Natl Acad Sci USA 1989; 86: 6508-12.
• 17 Eaton D, Rodriguez H, Vehar GA. Proteolytic processing of human factor VIII. Correlation of specific cleavages by thrombin, factor Xa and activated protein C with activation and inactivation of factor VIII coagulant activity. Biochem 1986; 25: 505-12.
• 18 Burnet FM. A modification of Jerne`s theory of antibody production using the concept of clonal selection. Aust J Sci 1957; 20: 67-9.
• 19 Bretscher P, Cohn M. A theory of self-nonself discrimination. Science 1970; 169: 1042-9.
• 20 Matzinger P. Tolerance, danger, and the extended family. Annu Rev Immunol 1994; 12: 991-1045.
• 21 Fay PJ. Activation of factor VIII and mechanisms of cofactor action. Blood Rev 2004; 18: 1-15.
• 22 Peerlinck K, Arnout J, Gilles JG. et al. A higher than expected incidence of factor VIII inhibitors in multitransfused haemophilia A patients treated with an intermediate purity pasteurized fector VIII concentrate. Thromb Haemost 1993; 69: 115-8.
• 23 Peerlinck K, Arnout J, Di Giambattista M. et al. Factor VIII inhibitors in previously treated hemophilia A patients with a double virus-inactivated plasma derived factor VIII concentrate. Thromb Haemost 1997; 77: 80-6.
• 24 Rock KL, Hearn A, Chen C-J. et al. Natural endogenous adjuvants. Springer Semin Immun 2005; 26: 231-46.
• 25 Basu S, Binder RJ, Suto R. et al. Necrotic but not apoptotic cell death releases heat shock proteins, which deliver a partial maturation signal to dendritic cells and activate the NF-kappa B pathway. Int Immunol 2000; 12: 1539-46.
• 26 Pulendran B. Immune activation: death, danger and dendritic cells. Current Biol 2004; 14: R30-2.
• 27 Lazarowski ER, Homolya L, Boucher RC. et al. Direct demonstration of mechanically induced release of cellular UTP and its implication for uridine nucleotide receptor activation. J Biol Chem 1997; 272: 24348-54.
• 28 Shi Y, Evans JE, Rock KL. Molecular identification of a danger signal that alerts the immune system to dying cells. Nature 2003; 425: 516-21.
• 29 Okamura Y, Watari M, Jerud ES. et al. The extra domain A of fibronectin activates Toll-like receptor 4. J Biol Chem 2001; 276: 10229-33.
• 30 Smiley ST, King JA, Hancock WW. Fibrinogen stimulates macrophage chemokine secretion through toll-like receptor 4. J Immunol 2001; 167: 2887-94.
• 31 Termeer C, Benedix F, Sleeman J. et al. Oligosaccharides of Hyaluronan activate dendritic cells via toll-like receptor 4. J Exp Med 2002; 195: 99-111.
• 32 Guillot L, Balloy V, McCormack FX. et al. Cutting edge: the immunostimulatory activity of the lung surfactant protein-A involves toll-like receptor 4. J Immunol 2002; 168: 5989-92.
• 33 Steinhoff M, Buddenkotte J, Shpacovitch V. et al. Proteinase-activated receptors: transducers of proteinase-mediated signalling in inflammation and immune response. Endocr Rev 2005; 26: 1-43.