Download PDF
Thromb Haemost 1996; 76(01): 017-022
DOI: 10.1055/s-0038-1650515
Original Article
Schattauer GmbH Stuttgart

T Lymphocyte Proliferative Responses Induced by Recombinant Factor VIII in Hemophilia A Patients with Inhibitors

Sylvia T Singer
1   The Hematology-Oncology Department, Children’s Hospital Oakland, Oakland, CA, USA
,
Joseph E Addiego Jr
1   The Hematology-Oncology Department, Children’s Hospital Oakland, Oakland, CA, USA
,
Donald C Reason
2   The Children’s Hospital Oakland Research Institute, Oakland, CA, USA
,
Alexander H Lucas
2   The Children’s Hospital Oakland Research Institute, Oakland, CA, USA
› Author Affiliations
Further Information

Publication History

Received: 06 November 1995

Accepted after resubmission09 April 1996

Publication Date:
26 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

In this study we sought to determine whether factor VUI-reactive T lymphocytes were present in hemophilia A patients with inhibitor antibodies. Peripheral blood mononuclear cells (MNC) were obtained from 12 severe hemophilia A patients having high titer inhibitors, 4 severe hemophilia A patients without inhibitors and 5 normal male subjects. B cell-depleted MNC were cultured in serum-free medium in the absence or presence of 2 µg of recombinant human factor VIII (rFVIII) per ml, and cellular proliferation was assessed after 5 days of culture by measuring 3H-thymidine incorporation. rFVIII induced marked cellular proliferation in cultures of 4 of 12 inhibitor-positive hemophilia patients: fold increase over background (stimulation index, SI) of 7.8 to 23.3. The remaining 8 inhibitor-positive patients, the 4 hemophilia patients without inhibitors and the 5 normal subjects, all had lower proliferative responses to rFVIII, SI range = 1.6 to 6.0. As a group, the inhibitor-positive subjects had significantly higher proliferative responses to rFVIII than did the inhibitor-negative and normal subjects (p < 0.05 by t-test). Cell fractionation experiments showed that T lymphocytes were the rFVIII-responsive cell type, and that monocytes were required for T cell proliferation. Thus, rFVIII-reactive T lymphocytes are present in the peripheral circulation of some inhibitor-positive hemophilia A patients. These T cells may recognize FVIII in an antigen-specific manner and play a central role in the regulation of inhibitor antibody production

 

  • References

  • 1 Hoyer LW. Hemophilia A. N Engl J Med 1994; 330: 38-47
    CrossrefPubMedGoogle Scholar
  • 2 Ehrenforth S, Kreuz W, Scharrer I, Linde R, Funk M, Giingor T, Krackhardt B, Kornhuber B. Incidence of development of factor VIII and factor IX inhibitors in haemophiliacs. Lancet 1992; 339: 594-598
    CrossrefPubMedGoogle Scholar
  • 3 Addiego J, Kasper C, Abildgaard C, Hilgartner M, Lusher J, Glader B, Aledort L. Frequency of inhibitor development in haemophiliacs treated with low-purity factor VIII. Lancet 1993; 342: 462-464
    CrossrefPubMedGoogle Scholar
  • 4 Lusher J, Arkin S, Abildgaard CF, Hurst D. the Kogenate PUP Study Group and other participating institutions. Kogenate treatment of previously untreated patients (PUPS) with hemophilia A: Update of safety, efficacy, and inhibitor development after five study years. Blood 1994; (abstr) 84: 239a
    PubMedGoogle Scholar
  • 5 Kasper CK. Treatment of factor VIII inhibitors. Prog Hemost Thromb 1989; 9: 57-86
    PubMedGoogle Scholar
  • 6 Lubahn BC, Reisner HM. Characterization of a monoclonal anti-idiotype antibody to human anti-factor VIII antibodies. Proc Natl Acad Sci USA 1990; 87: 8232-8236
    CrossrefPubMedGoogle Scholar
  • 7 Scandella D, Mattingly M, de Graaf S, Fulcher CA. Localization of epitopes for human factor VIII inhibitor antibodies by immunoblotting and antibody neutralization. Blood 1989; 74: 1618-1626
    PubMedGoogle Scholar
  • 8 Arai MD, Scandella D, Hoyer LW. Molecular basis of factor VIII inhibition by human antibodies: Antibodies that bind to the factor VIII light chain prevent the interaction of factor VIII with phospholipid. J Clin Invest 1989; 83: 1978-1984
    CrossrefPubMedGoogle Scholar
  • 9 Foster PA, Fulcher CA, Houghten RA, Mahoney de SG, Zimmerman TS. Localization of the binding regions of a murine monoclonal anti-factor VIII antibody and a human anti-factor VIII alloantibody, both of which inhibit factor VIII procoagulant activity, to amino acid residues threonine351-serine365 of the factor VIII heavy chain. J Clin Invest 1988; 82: 123-128
    CrossrefPubMedGoogle Scholar
  • 10 Lollar P, Parker ET, Curtis JE, Helgerson SL, Hoyer LW, Scott ME, Scandella D. Inhibition of human factor Villa by anti-A2 subunit antibodies. J Clin Invest 1994; 93: 2497-2504
    CrossrefPubMedGoogle Scholar
  • 11 Fulcher CA, Mahonney de SG, Zimmerman TS. FVIII inhibitor IgG subclass and FVIII polypeptide specificity determined by immunoblotting. Blood 1987; 69: 1475-1480
    PubMedGoogle Scholar
  • 12 Scandella D, Nakai H, Felch M, Prescott R, Bray G, Hurst D. and the Recombinate and Kogenate Study Groups. Determination of anti-factor VIII antibodies responsible for the inhibitor titer in hemophiliacs treated with plasma or recombinant factor VIII and in spontaneous inhibitors. Blood 1994; (abstr) 84: 240a
    PubMedGoogle Scholar
  • 13 Saenko EL, Shima M, Rajalakshmi KJ, Scandella D. A role for the C2 domain of factor VIII in binding to von Willebrand factor. J Biol Chem 1994; 269: 11601-11605
    PubMedGoogle Scholar
  • 14 Rizza CR, Matthews JM. Effect of frequent factor VIII replacement on the level of factor VIII antibodies in haemophiliacs. Br J Hematol 1982; 52: 13-24
    CrossrefPubMedGoogle Scholar
  • 15 Shapiro SS. Genetic predisposition to inhibitor formation. In: Factor VIII Inhibitors Hoyer LW. (ed) New York, NY, Alan R. Liss, Inc; 1984: 45-56
  • 16 Aly AM, Aledort LM, Lee TD, Hoyer LW. Histocompatibility antigen patterns in haemophilic patients with factor VIII antibodies. Br J Haematol 1990; 76: 238-241
    CrossrefPubMedGoogle Scholar
  • 17 Tuddenham EGD, Schwaab R, Seehafer J, Millar DS, Gitschier J, Higuchi M, Bidichandani S, Connor JM, Hoyer LW, Yoshioka A, Peake IR, Olek K, Kazazian HH, Lavergne JM, Giannelli Antonarakis SE, Cooper DN, Hemophilia A. database of nucleotide substitutions, deletions, insertions and rearrangements of the factor VIII gene, second edition. Nucleic Acids Res 1994; 22: 4851-4868
    CrossrefPubMedGoogle Scholar
  • 18 Figueiredo MS, Bemardi F, Zago MA. A novel deletion of FVIII gene associated with variable levels of FVIII inhibitor. Eur J Haematol 1992; 48: 152-154
    PubMedGoogle Scholar
  • 19 Lakich D, Kazazian Jr HH, Antonarakis SE, Gitschier J. Inversions disrupting the factor VIII gene are a common cause of severe haemophilia A. Nature Genetics 1993; 5: 236-241
    CrossrefPubMedGoogle Scholar
  • 20 Goodeve AC, Preston FE, Peake IR. Factor VIII gene rearrangements in patients with severe hemophilia A. Lancet 1994; 343: 329-330
    CrossrefPubMedGoogle Scholar
  • 21 Antonarakis SE, Rossiter JP, Young M, Horst J, de Moerloose P, Sommer SS, Ketterling RP, Kazazian Jr HH, Negrier C, Vinciguerra C, Gitschier J, Goossens M, Girodon E, Ghanem N, Plassa F, Lavergne JM, Vidaud M, Costa JM, Laurian Y, Lin SW, Lin SR, Shen MC, Lillicrap D, Taylor SAM, Windsor S, Valleix SV, Nafa K, Sultan Y, Delpech M, Vnencak-Jones CL, Phillips JA, Ljung RCR, Koumbarelis E, Gialeraki A, Mandalaki T, Jenkins PV, Collins PW, Pasi KJ, Goodeve A, Peake I, Preston FE, Schwartz M, Scheibel E, Ingerslev J, Cooper DN, Millar DS, Kakkar VV, Giannelli F, Naylor JA, Tizzano EF, Baiget M, Domenech M, Altisent C, Tusell J, Beneyto M, Lorenzo JI, Gaucher C, Mazurier C, Peerlinck K, Matthijs G, Cassiman JJ, Vermylen J, Mori PG, Acquila M, Caprino D, Inaba H. Factor VIII gene inversions in severe hemophilia A: results of an international consortium study. Blood 1995; 86: 2206-2212
    PubMedGoogle Scholar
  • 22 Millar DS, Kakkar VV, Cooper DN. Screening for inversions in the factor VIII (F8) gene causing severe hemophilia A. Blood Coag Fibrinol 1994; 5: 239-242
    CrossrefPubMedGoogle Scholar
  • 23 Frommel D, Allain JP. Genetic predisposition to develop factor VIII antibody in classic hemophilia. Clin Immunol Immunopathol 1977; 8: 34-38
    CrossrefPubMedGoogle Scholar
  • 24 Addiego JE, Kasper C, Abildgaard C, Lusher J, Hilgartner M, Glader B, Aledort L, Hurst D, Bray G. Increased frequency of inhibitors in African American hemophilia A patients. Blood 1994; (abstr) 84: 239a
    PubMedGoogle Scholar
  • 25 Allain JP, Verroust F. The response to factor VIII infusion in inhibitor patients. In: Factor Vlll Inhibitors. Hoyer LW, ed, New York, NY, Alan R. Liss, Inc 1984: 99-108
    PubMedGoogle Scholar
  • 26 Littlewood JD, Bevan SA, Kemball-Cook G, Evans RJ, Barrowcliffe TW. Variable inactivation of human factor VIII from different sources by human factor VIII inhibitors. Br J Haematol 1991; 77: 535-538
    PubMedGoogle Scholar
  • 27 Rosendaal FR, Nieuwenhuis HK, van den Berg HM, Heijboer H, Mauser-Bunschoten EP, van der Meer J, Smit C, Strengers PFW, Briet E. and the Dutch Hemophilia Study Group. A sudden increase in factor VIII inhibitor development in multitransfused hemophilia A patients in the Netherlands. Blood 1993; 81: 2180-2186
    PubMedGoogle Scholar
  • 28 Kasper CK, Aledort LM, Counts RB. A more uniform measure of factor VIII inhibitors. Thromb Diath Haemorrh 1975; 34: 869-872
    PubMedGoogle Scholar
  • 29 Bayer AG. Germany. Recombinant factor VIII. Proceedings of the First International Symposium on Recombinant Factor VIII held on August 26, 1989 in Tokyo, Japan. Seminars in Hematology 1991; 28: 1
    PubMedGoogle Scholar
  • 30 Weiner MDS, Bianco C, Nussenzweig V. Enhanced binding of neuraminidase-treated sheep erythrocytes to human T lymphocytes. Blood 1973; 42: 939-946
    PubMedGoogle Scholar
  • 31 Madhok R, Smith J, Jenkins A, Lowe DO. T cell sensitization to factor VIII in haemophilia A. Br J Haem 1991; 79: 235-238
    CrossrefPubMedGoogle Scholar
  • 32 Bray GL, Kroner BL, Arkin S, Aledort LW, Hilgartner MW, Eyster ME, Ragni MV, Goedert JJ. Loss of high-responder inhibitors in patients with severe hemophilia A and human immunodeficiency virus type 1 infection: A report from the multi-center hemophilia cohort study. Am J Hem 1993; 42: 375-379
    CrossrefPubMedGoogle Scholar
  • 33 Ragni MV, Bontempo FA, Lewis JH. Disappearance of inhibitor to factor VIII in HIV-infected hemophiliacs with progression to AIDS or severe ARC. Transfusion 1989; 29: 447-449
    CrossrefPubMedGoogle Scholar
  • 34 Leyva WH, Knusten AP, Joist JH. Disappearance of a high responder factor VIII inhibitor in a hemophiliac with AIDS. Am J Clin Pathol 1988; 89: 414-418
    CrossrefPubMedGoogle Scholar
  • 35 Algiman M, Dietrich G, Nydegger UE, Boieldieu D, Sultan Y, Kazatchkine MD. Natural antibodies to factor VIII (anti-hemophilic factor) in healthy individuals. Proc Natl Acad Sci USA 1992; 89: 3795-3799
    CrossrefPubMedGoogle Scholar
  • 36 Gilles JG, Saint-Remy JM. Healthy subjects produce both anti-factor VIII and specific anti-idiotypic antibodies. J Clin Invest 1994; 94: 1496-1505
    CrossrefPubMedGoogle Scholar
  • 37 Green D, Lechner K. A survey of 215 non-hemophilic patients with inhibitors to factor VIII. Thromb Haemost 1982; 45: 200-204
    Article in Thieme ConnectPubMedGoogle Scholar
  • 38 Lottenberg R, Kentro TB, Kitchens CS. Acquired hemophilia. A natural history study of 16 patients with factor VIII inhibitors receiving little or no therapy. Arch Intern Med 1987; 147: 1077-1081
    CrossrefPubMedGoogle Scholar
  • 39 Bums J, Rosenzweig A, Zweiman B, Lisak RB. Isolation of myelin basic protein-reactive T-cell lines from normal human blood. Cell Immunol 1983; 81: 435-440
    CrossrefPubMedGoogle Scholar
  • 40 Pette M, Fujita K, Wilkinson D, Altmann DM, Trowsdale J, Giegerich G, Hinkkanen A, Epplen JT, Kappos L, Wekerle H. Myelin autoreactivity in multiple sclerosis: Recognition of myelin basic protein in the context of HLA-DR2 products by T lymphocytes of multiple-sclerosis patients and healthy donors. Proc Natl Acad Sci USA 1990; 87: 7968-7972
    CrossrefPubMedGoogle Scholar
  • 41 Ota K, Matsui M, Milford EL, Mackin GA, Weiner HL, Hafler DA. T-cell recognition of an immunodominant myelin basic protein epitope in multiple sclerosis. Nature 1990; 346: 183-187
    CrossrefPubMedGoogle Scholar
  • 42 Filion M, Devine D, Decarv F, Chartrand P. In vitro presentation of platelet antigen GPIIB-IIIA initiates T-cell proliferation even with immuno-com-patible individual. Blood 1992; (abstr) 80: 49a
    PubMedGoogle Scholar
  • 43 Kuwana M, Medsger TA, Wright TM. T cell proliferative response induced by DNA topoisomerase I in patients with systemic sclerosis and healthy donors. J Clin Invest 1995; 96: 586-596
    CrossrefPubMedGoogle Scholar
  • 44 Tordai A, Fenton JW II, Andersen T, Gelfand EW. Functional thrombin receptors on human T lymphoblastoid cells. J Immunol 1993; 150: 4876-4886
    PubMedGoogle Scholar
  • 45 Tiarks C, Humphreys RE, Anderson J, Mole J, Pechet L. Hypothesis for the control of clotting factor VIII inhibitory antibodies by decreasing potency of helper T-cell-recognized epitopes in factor VIII. Scand J Immunol 1992; 36: 653-660
    CrossrefPubMedGoogle Scholar
  • 46 Hoyer LW. Future approaches to factor VIII inhibitor therapy. Amer J Med 1991; 91 (05) 40S-47S
    PubMedGoogle Scholar