Abstract
In chronic idiopathic thrombocytopenic purpura (ITP) platelet destruction is caused by antibodies directed against platelet membrane glycoproteins (GP), and the predominant autoantigens are known to be GPIb/IX and GPIIb/IIIa. In a recent study we reported that these antibodies frequently had a restricted light chain phenotype, thereby supporting a clonal origin. Similar findings and the presence of clonal B-cell populations in immune thrombocytopenias have been reported by others. In the present study we further explored the hypothesis of clonal B-cell expansions in chronic ITP. Twenty patients with chronic ITP were investigated. Antibodies were detected with an ELISA (MAIPA) specific for GPIb/IX and GPIIb/IIIa; circulating clonal B lymphocytes were assessed by flow-cytometric (FACE) clonal-excess analysis and by analyzing Ig-gene rearrangements (CDR3) with the PCR technique. Nine patients displayed a GP-specific antibody restricted to either kappa or lambda phenotype. However, FACS analysis and Ig-gene rearrangement studies did not disclose any circulating clonal B-cell population. Considering the sensitivity of the FACS analysis and Ig-gene rearrangement for detection of clonal B-cell populations, the hypothesis of clonally derived autoantibodies in ITP is still valid. Most probably, the clonal B-cell expansion responsible for the production of autoantibodies in ITP, if present, is below the detection limit for the techniques employed.
Similar content being viewed by others
References
Agrawal YP, Hämäläinen E, Mahlamäki EK, Aho H, Nousiainen T, Lahtinen R, Penttilä IM (1992) Comparison of poly- and monoclonal antibodies for determination of B-cell clonal excess in a routine clinical laboratory. Eur J Haematol 48:49–55
Beardsley DS, Spiegel JE, Jacobs MM, Handin RI, Lux SE (1984) Platelet membrane glycoprotein IIIa contains target antigens that bind anti-platelet antibodies in immune thrombocytopenias. J Clin Invest 74:1701–1707
Billadeau D, Blackstadt M, Greipp P, Kyle RA, Oken MM, Kay N, van Ness B (1991) Analysis of B-lymphoid malignancies using allele-specific polymerase chain reaction: a technique for sequential quantitation of residual disease. Blood 78:3021–3029
Brisco MJ, Tan LW, Orsborn AM, Morley AA (1990) Development of a highly sensitive assay, based on the polymerase chain reaction, for rare B-lymphocyte clones in a polyclonal population. Br J Haematol 75:163–167
Carson DAN, Chen PP, Kipps TJ, Radoux V, Jirik FR, Goldfien RD, Fox RJ, Silverman GJ, Fong S (1987) Idiotypic and genetic studies of human rheumatoid factors. Arthritis Rheum 30:1321–1325
Christie DJ, Sauro SC, Fairbanks KD, Kay NE (1993) Detection of clonal platelet antibodies in immunologically mediated thrombocytopenias: association with circulating clonal/ oligoclonal B cells. Br J Haematol 85:277–284
van der Harst D; de Jong D, Limpens J, Kluin PhM, Rozier Y, van Ommen GJB, Brand A (1990) Clonal B-cell populations in patients with idiopathic thrombocytopenic purpura. Blood 76:2321–2326
Hewitt J, Burton IF (1994) Incidence of autoantibodies to GPIIb/IIIa in chronic autoimmune thrombocytopenic purpura may be overestimated by the MAIPA. Br J Haematol 86:418–420
Hiruiwa A, Nugent D, Miller ECB (1990) Sequence analysis of monoclonal antibodies derived from a patient with idiopathic thrombocytopenic purpura. Autoimmunity 8:107–113
Hymes KB, Karpatkin S (1990) In vitro suppressor T lymphocyte dysfunction in autoimmune thrombocytopenic purpura associated with a complement-fixing antibody. Br J Haematol 74:330–335
Johnson A, Olofsson T (1993) Flow cytometric Conal excess analysis of peripheral blood, routine handling, and pitfalls in interpretation. Cytometry 14:188–195
Kekomaki R, Dawson B, McFarland J, Kunicki TJ (1991) Localization of human platelet autoantigens to the cystein-rich region of glycoprotein IIIa. J Clin Invest 88:847–854
Kiefel V, Santoso S, Weisheit M, Mueller-Eckhardt C (1987) Monoclonal antibody-specific immobilization of platelet antigens (MAIPA): a new tool for the identification of platelet-reactive antibodies. Blood 70:1722–1726
Kiefel V, Santoso S, Kaufmann E, Mueller-Eckhardt C (1991) Autoantibodies against platelet glycoprotein Ib/IX: a frequent finding in autoimmune thrombocytopenic purpura. Br J Haematol 79:256–262
Kiefel V, Mueller-Eckhardt C (1993) Report on the Fifth International Society of Blood Transfusion Platelet Serology Workshop. Transfusion 33:65–69
Lauria F, Mantovani V, Catosky D, Gaurini A, Gugliotta L, Mirone E, Tura S (1981) T gamma cell deficiency in idiopathic thrombocytopenic purpura (ITP). Scand J Haematol 26:156–160
Lauria F, Raspadori D, Gobbi M, Gaurini A, Gugliotta L, Aieta M, Tura S (1983) T-cell subsets defined by monoclonal antibodies in autoimmune thrombocytopenic purpura (ATP). Haematologica (Pavia) 68:600–606
van Leeuwen EFJ, van der Venth M, Engelfreit CP, von dem Borne AEGK (1982) Specificity of autoantibodies in autoimmune thrombocytopenia. Blood 59:23–26
Loken MR, Brosnan JM, Bach BA, Ault KA (1990) Establishing optimal lymphocyte gates for immunophenotyping by flow Cyometry. Cytometry 11:453–459
Miller SA, Dykes DD, Polesky HF (1988) A simple salting out procedure for extracting DNA from nucleated cells. Nucleic Acids Res 16:1215–1218
Misutani H, Katagiri S, Uejima K, Ohnishi M, Tamaki T, Kanayama Y, Tsubakio T, Kurata Y, Yonezawa T, Tarui S (1985) T-cell abnormalities in patients with idiopathic thrombocytopenic purpura: the presence of OKT4+8+ cells. Scand J Haematol 35:233–239
Mylvaganan R, Ahn YS, Harrington WJ, Kim CL, Gratzner HG (1985) Differences in T cell subsets between men and women with idiopathic thrombocytopenic purpura. Blood 66:967–972
Nugent DJ (1989) Human monoclonal antibodies in the characterization of platelet antigens. In: Kunicki TJ, George JN (eds) Platelet immunobiology: molecular and clinical aspects. Lippincott, Philadelphia, pp 273
Peller S, Kaufman S, Shaked U (1986) Immuno-regulatory defect in patients with autoimmune thrombocytopenic purpura (ATP). Thymus 8:361–367
Scott CS, Wheeler R, Ford P, Bynoe AG, Roberts BE (1983) T lymphocyte subpopulations in idiopathic thrombocytopenic purpura (ITP). Scand J Haematol 40:401–406
Semple JW, Freedman J (1991) Increased antiplatelet T helper lymphocyte reactivity in patients with autoimmune thrombocytopenia. Blood 78:2619–2625
Semple JW, Freedman J (1992) Cellular immune mechanisms in chronic autoimmune thrombocytopenic purpura (ATP). Autoimmunity 13:311–319
Silberstein LE, Jefferies LC, Goldman J, Friedman D, Moore JS, Nowell PC, Roeleke D, Pruzanski W, Roudier J, Silverman GC (1991) Variable region gene analysis of pathologic human autoantibodies to the related i and I red blood cell antigens. Blood 78:2372–2386
Sthoeger ZM, Sthoeger D, Shtalrid M, Sigler E, Geltner D, Berrebi A (1993) Mechanism of autoimmune hemolytic anemia in chronic lymphocyte leukemia. Am J Hematol 43:259–264
Stockelberg D, Jacobsson S, Kutti J, Wadenvik H (1995) Evidence for a light chain restriction of glycoprotein Ib/IX and IIb/IIIa reactive autoantibodies in chronic immune thrombocytopenic purpura. Br J Haematol 90:175–179
Szatkowski NS, Kunicki TJ, Aster RH (1986) Identification of glycoprotein Ib as a target for autoantibody in idiopathic (autoimmune) thrombocytopenic purpura. Blood 67:310–315
Trainor KJ, Brisco MJ, Colin JS, Alexander AM (1990) Monoclonality in B-lymphoproliferative disorders detected at the DNA level. Blood 11:2220–2222
Trent R, Adams E, Erhardt C, Basten A (1981) Alterations in T-gamma cells in patients with chronic idiopathic thrombocytopenic purpura. J Immunol 127:621–626
Varon D, Karpatkin S (1983) A monoclonal anti-platelet antibody with decreased reactivity for autoimmune thrombocytopenic platelets. Proc Natl Acad Sci USA 80:6992–6995
Woods VL, Oh E, Mason D, McMillan R (1984) Autoantibodies against the platelet glycoprotein IIb/IIIa complex in patients with chronic ITP. Blood 63:368–375
Woods VL, Kurata Y, Montgomery RR, Tani P, Mason D, Oh E, McMillan R (1984) Autoantibodies against platelet glycoprotein Ib in patients with chronic immune thrombocytopenic purpura. Blood 64:156–160
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Stockelberg, D., Hou, M., Jacobsson, S. et al. Light chain-restricted autoantibodies in chronic idiopathic thrombocytopenic purpura, but no evidence for circulating clonal B-lymphocytes. Ann Hematol 72, 29–34 (1996). https://doi.org/10.1007/BF00663013
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00663013