Download PDF
Thromb Haemost 2007; 97(05): 788-794
DOI: 10.1160/TH06-09-0519
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Cytokine production by CD4+ T cells specific for coagulation factor VIII in healthy subjects and haemophilia A patients

Genlin Hu
1   Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA
,
Delan Guo
1   Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA
,
Nigel S. Key
2   Department of Medicine, Division of Hematology/Oncology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA
,
Bianca M. Conti-Fine
1   Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota, USA
› Author AffiliationsFinancial support: This study was supported by the NHLBI grant HL61922 (to B.M.C.-F.) and HL65578 (to N.S.K).
Further Information

Publication History

Received 13 September 2006

Accepted after resubmission 01 March 2007

Publication Date:
24 November 2017 (online)

    Permissions and Reprints

Summary

HaemophiliaA patients treated with human factorVIII (fVIII) may develop antibody (Ab) inhibitors to fVIII. FVIII-specific CD4+ T cells are common in haemophilia A patients, but also in healthy subjects who do not have a sustained anti-fVIII Ab response. Here, we examined the fVIII-induced IFNγ -, IL-4- and TGF- β 1-producing CD4+ T blasts by culturing peripheral blood mononuclear cells (PBMC) from controls and patients with recombinant fVIII. FVIII exposure significantly increased IFNγ - and IL-4-, but not TGF-β 1-producing CD4+ T blasts in patients with inhibitors. Patients without inhibitors had fVIII-induced IFNγ - andTGF-β 1-,but not IL-4-producing CD4+ T blasts.Controls did not have IL-4-producing CD4+ T blasts. However, controls whose PMBC proliferated in response to fVIII had fVIII-induced CD4+ T blasts that produced IFN-γ, the number of which correlated with the intensity of the proliferative response to fVIII of their PMBC, whereas controls whose PMBC did not proliferate to fVIII had predominantly fVIII-induced CD4+ T blasts that producedTGF- β 1.The presence in controls and patients without inhibitors of fVIII-induced IFN-γ -producing CD4 + T cells, but not IL-4-producing CD4+ T cells, which are abundant in inhibitor patients, suggests a role of Th1 cells in initiating the immune response to fVIII, and of Th2 cells in the development of strong inhibitor production. The polarized high ratios of Th3/Th1 and Th3/Th2 in controls and patients without inhibitors suggest that a preponderance ofTh3 cells in the response to fVIII may help to maintain tolerance to fVIII.

Keywords

CD4+ T cells - cytokines - factor VIII - hemophilia A - inhibitors
 

  • References

  • 1 Key NS. Inhibitors in congenital coagulation disorders. Br J Haematol 2004; 127: 379-391.
    CrossrefPubMedGoogle Scholar
  • 2 Franchini M, Salvagno GL, Lippi G. Inhibitors in mild/moderate haemophilia A: an update. Thromb Haemost 2006; 96: 113-118.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Reding MT, Wu HY, Krampf M. et al. Sensitization of CD4+ T cells to coagulation factor VIII: response in congenital and acquired hemophilia patients and in healthy subjects. Thromb Haemost 2000; 84: 634-652.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Singer ST, Addiego JE, Reason DC. et al. T lymphocyte proliferative responses induced by recombinant factor VIII in hemophilia A patients with inhibitors. Thromb Haemost 1996; 76: 17-22.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Hu GL, Okita DK, Diethelm-Okita BM. et al. Recognition of coagulation factor VIII by CD4+ T cells of healthy human. J Thromb Haemost 2003; 01: 2159-2166.
    CrossrefPubMedGoogle Scholar
  • 6 Moreau A, Lacroix-Desmazes S, Stieltjes N. et al. Antibodies to the fVIII light chain that neutralize FVIII procoagulant activity are present in plasma of nonresponder patients with severe hemophilia A and normal polyclonal human IgG. Blood 2000; 95: 3435-3441.
    PubMedGoogle Scholar
  • 7 Batlle J, Gomez E, Rendal E. et al. Ab to factor VIII in plasma of patients with hemophilia A and normal subjects. Ann Hematol 1996; 72: 321-326.
    CrossrefPubMedGoogle Scholar
  • 8 Gilles JGG, Saint-Remy JMR. Healthy subjects produce both anti-factor VIII and specific anti-idiotypic antibodies. J Clin Invest 1994; 94: 1496-1505.
    CrossrefPubMedGoogle Scholar
  • 9 Murphy KM. T lymphocyte differentiation in the periphery. Curr Opin Immunol 1998; 10: 226-232.
    CrossrefPubMedGoogle Scholar
  • 10 Santana MA, Rosenstein Y.. What it takes to become an effector T cell: the process, the cells involved, and the mechanisms. J Cell Physiol 2003; 195: 392-401.
    CrossrefPubMedGoogle Scholar
  • 11 Mills KH. Antigen-specific regulatory T cells-- their induction and role in infection. Semin Immunol 2004; 16: 107-117.
    CrossrefPubMedGoogle Scholar
  • 12 Maggi E, Cosmi L, Liotta F. et al. Thymic regulatory T cells. Autoimmun Rev 2005; 04: 579-586.
    CrossrefPubMedGoogle Scholar
  • 13 Weiner HL. Induction and mechanism of action of transforming growth factor- β -secreting Th3 regulatory cells. Immunol Rev 2001; 182: 207-214.
    CrossrefPubMedGoogle Scholar
  • 14 Lan RY, Ansari AA, Lian ZX. et al. Regulatory T cells: development, function and role in autoimmunity. Autoimmun Rev 2005; 04: 351-363.
    CrossrefPubMedGoogle Scholar
  • 15 Holzer U, Kwok WW, Nepom GT. et al. Differential antigen sensitivity and costimulatory requirements in human Th1 and Th2 antigen-specific CD4+ cells with similar TCR avidity. J Immunol 2003; 170: 1218-1223.
    CrossrefPubMedGoogle Scholar
  • 16 Valentini G, Baroni A, Esposito K. et al. Peripheral blood T lymphocytes from systemic sclerosis patients show both Th1 and Th2 activation. J Clin Immunol 2001; 21: 210-217.
    CrossrefPubMedGoogle Scholar
  • 17 Reding MT, Lei S, Lei H. et al. Distribution of Th1- and Th2-induced anti-factor VIII IgG subclasses in congenital and acquired hemophilia patients. Thromb Haemost 2002; 88: 568-575.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Perez-Machadol MA, Ashwoodl P, Thomson MA. et al. Reduced transforming growth factor- β 1-producing T cells in the duodenal mucosa of children with food allergy. Eur J Immunol 2003; 33: 2307-2315.
    CrossrefPubMedGoogle Scholar
  • 19 Chatenoud L, Bach LP. Adaptive human regulatory T cells: myth or reality?. J Clin Invest 2006; 116: 2325-2327.
    CrossrefPubMedGoogle Scholar
  • 20 Kitani A, Chua K, Nakamura K. et al. Activated self-MHC-reactive T cells have the cytokine phenotype of Th3/T regulatory cell 1 T cells. J Immunol 2000; 165: 691-70.
    CrossrefPubMedGoogle Scholar
  • 21 Kim YK, Myint AM, Lee BH. et al. Th1, Th2 and Th3 cytokine alteration in schizophrenia. Prog Neuropsychopharmacol Biol Psychiatry 2004; 28: 1129-1134.
    CrossrefPubMedGoogle Scholar
  • 22 Akbari O, Stock P, DeKruyff RH. et al. Role of regulatory T cells in allergy and asthma. Curr Opin Immunol 2003; 15: 627-633.
    CrossrefPubMedGoogle Scholar
  • 23 Abbas A, Murphy K, Sher A.. Functional diversity of helper T cell lymphocytes. Nature 1996; 383: 787-793.
    CrossrefPubMedGoogle Scholar
  • 24 Seder RA, Paul WE. Acquisition of lymphokineproducing phenotype by CD4+ T cells. Ann Rev Immunol 1994; 12: 635-673.
    CrossrefPubMedGoogle Scholar
  • 25 Gilles JGG, Jacquemin MG, Saint-Remy JMR. Factor VIII inhibitors. Thromb Haemost 1997; 78: 641-646.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 26 Fulcher CA, de Graaf Mahoney S, Zimmerman TS. FVIII inhibitor IgG subclass and FVIII polypeptide specificity determined by immunoblotting. Blood 1987; 69: 1475-1480.
    PubMedGoogle Scholar
  • 27 Wu H, Reding M, Qian J. et al. Mechanism of the immune response to human factor VIII in murine hemophilia A. Thromb Haemost 2001; 85: 125-133.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 28 Bril WS, van Helden PM, Hausl C. et al. Tolerance to factor VIII in a transgenic mouse expressing human factor VIII cDNA carrying an Arg(593) to Cys substitution. Thromb Haemost 2006; 95: 341-347.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Reipert BM, Ahmad RU, Turecek PL. et al. Characterization of antibodies induced by human factor VIII in a murine knockout model of hemophilia A. Thromb Haemost 2000; 84: 826-832.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Gomes JAS, Bahia-Oliveira LMG, Rocha MOC. et al. Evidence that development of severe cardiomyopathy in human Chagas’disease is due to a Th1-specific immune response. Infect Immun 2003; 71: 1185-1193.
    CrossrefPubMedGoogle Scholar
  • 31 Seder RA, Marth T, Sieve MC. et al. Factors involved in the differentiation of TGF- β -producing cells from naive CD4+ T cells: IL-4 and IFN-γ have opposing effects, while TGF- β positively regulates its own production. J Immunol 1998; 160: 5719-5728.
    PubMedGoogle Scholar
  • 32 Pala P, Hussell T, Openshaw PJM. Flow cytometric measurement of intracellular cytokines. J Immunol Methods 2000; 243: 107-124.
    CrossrefPubMedGoogle Scholar
  • 33 Cao O, Armstrong E, Schlachterman A. et al. Immune deviation by mucosal antigen administration suppresses gene-transfer-induced inhibitor formation to factor IX. Blood 2006; 108: 480-486.
    CrossrefPubMedGoogle Scholar
  • 34 Im SH, Barchan D, Souroujon MC. et al. Role of tolerogen conformation in induction of oral tolerance in experimental autoimmune myasthenia gravis. J Immunol 2000; 165: 3599-3605.
    CrossrefPubMedGoogle Scholar
  • 35 Miller A, Shapiro S, Gershtein R. et al. Treatment of multiple sclerosis with Copolymer-1: implicating mechanisms of Th1 to Th2/Th3 immune-deviation. J Neuroimmunol 1998; 92: 113-121.
    CrossrefPubMedGoogle Scholar
  • 36 Sprent J, Surh CD. T cell memory. Ann Rev Immunol 2002; 20: 551-579.
    CrossrefPubMedGoogle Scholar
  • 37 Constant S L, Bottomly K. Induction of Th1 and Th2 CD4+ T cell responses: the alternative approach. Ann Rev Immunol 1997; 15: 297-332.
    CrossrefPubMedGoogle Scholar