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Expansion of a B-lymphocyte clone producing IgM auto-antibodies encoded by a somatically mutated VHI gene in the spleen of an autoimmune patient

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Summary

Seventy-six human B-cell hybridomas were obtained by fusing B lymphocytes from the spleen of a patient with chronic idiopathic thrombocytopenia (ITP). Two independent hybridoma clones producing IgM autoantibodies reacting with platelets and other antigens from both the internal and the external environments were established from this fusion experiment. The IgM autoantibodies produced by the two hybridoma clones were found to be encoded by identical VHDJH and VLJL genes. The comparison of the VHI gene expressed in both hybridomas with the germline equivalent cloned from the patient's DNA showed two somatic mutations in the complementarity-determining regions CDR1 and CDR2 resulting in amino acid replacements. These data suggest the selection and expansion of an autoantibody-producing B-cell clone in the spleen of an ITP patient, probably as a result of (auto)antigen-driven stimulation.

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References

  1. Tan EM (1989) Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol 44:93–151

    PubMed  Google Scholar 

  2. Swaak AJG, Aarden LA, Statius van Eps LW, Feltkamp TEW (1979) Anti-dsDNA and complement profiles as prognostic guides in SLE. Arthritis Rheum 22:226–232

    PubMed  Google Scholar 

  3. Winfield JB, Faiferman I, Kofler D (1977), Avidity of anti-DNA antibodies in serum and IgG glomerular eluates from patients with systemic lupus erythematosus: association of high avidity anti-native DNA antibody with glomerulo-nephritis. J Clin Invest 59:90–97

    PubMed  Google Scholar 

  4. Watts RA, Ravirajan CT, Wilkinson LS, Williams W, Griffiths M, Butcher D, Horsfall AT, Staines AN, Isenberg DA (1991) Detection of human and murine common idiotypes of DNA antibodies in tissues and sera of patients with autoimmune diseases. Clin Exp Immunol 83:267–273

    PubMed  Google Scholar 

  5. Karpatkin S, Strick N, Karpatkin MB, Siskind GW (1972) Cumulative experience in the detection of antiplatelet antibody in 234 patients with idiopathic thrombocytopenic purpura, systemic lupus erythematosus and other clinical disorders. Am J Med 52:776–785

    Article  PubMed  Google Scholar 

  6. Buchanan GR (1989) Overview on ITP modalities in children. Blut 59:96–104

    PubMed  Google Scholar 

  7. Kunicki TJ, Furihata F, Kekomaki P, Scott JP, Nugent DJ (1990) A human monoclonal autoantibody specific for human platelet glycoprotein IIb (integrin α IIb) heavy chain. Hum Antibodies Hybridomas 1:83–89

    PubMed  Google Scholar 

  8. Kekomani R, Dawson B, McFarland J, Kunicki TJ (1991) Localization of human platelet autoantigens to the cysteinerich region of glycoprotein IIIa. J Clin Invest 88:847–854

    PubMed  Google Scholar 

  9. Lampman GW, Furie B, Schwartz RS, Stollar BD, Furie BC (1989) Amino acid sequence of a platelet-binding human anti-DNA monoclonal antibody. Blood 74:262–269

    PubMed  Google Scholar 

  10. Meng Q, Rauch J (1991) A systemic lupus erythematosusderived human hybridoma autoantibody reactive with antigens expressed on ADP-activated platelets. J Autoimmun 4:447–461

    Article  PubMed  Google Scholar 

  11. Zerella JT, Martin LW, Lampkin BC (1978) Emergency splenectomy for idiopathic thrombocytopenic purpura in children. J Pediatr Surg 13:243–246

    PubMed  Google Scholar 

  12. Skootsky SA, Rosove MH, Langley MB (1986) Immune thrombocytopenia and response to splenectomy in chronic liver diseases. Arch Intern Med 146:555–557

    Article  PubMed  Google Scholar 

  13. Merino J, Qin H, Schurmans S, Gretener D, Grau GE, Lambert PH (1989) Thrombocytopenia associated with the induction of neconatal tolerance to alloantigens: immunopathogenic mechanism. Eur J Immunol 19:1693–1699

    PubMed  Google Scholar 

  14. Theophilopoulos AN, Kofler R (1989) Molecular aspects of autoimmunity. Immunol Today 10:180–183

    Article  PubMed  Google Scholar 

  15. Steinberg AD, Krieg AM, Gorley MF, Klinman DM (1990) Theoretical and experimental approaches to generalized autoimmunity. Immunol Rev 118:129–163

    PubMed  Google Scholar 

  16. Jahn S, Grunow R, Kiessig ST, Bogacheva GT, Arsenjeva EL, Hlinka A, Rocklin OV, Baehr R von (1987) Cell biology of human IgM-producing hybridomas derived from a fusion with mouse myeloma cells. Hybridoma 1987; 6:679–687

    Google Scholar 

  17. Specht U, Mau H, Winter H, Jahn S, Volk HD, Cario WR (1988) Partial splenectomy for treatment of chronic idiopathic thrombocytopenia in childhood. Folia Haematol 115:509–514

    Google Scholar 

  18. Jahn S, Schwab J, Kiessig ST, Lukowsky A, Huhnholtz K, Mehl M, Specht U, Grunow R, Baehr R von (1989) Occurrence of hybridomas producing multispecific IgM in human-(humanmouse) fusions with lymphocytes from different human immune compartments. Allerg Immunol 35:271–277

    Google Scholar 

  19. Uchiyama H, Akatsuka J (1989) Elisa-plaque assay for the detection of platelet antibody-secreting cells. I. Studies on optimizing of reagent concentrations and culture conditions for enumerating of spot-forming cells. J Immunol Meth 117:199–203

    Article  Google Scholar 

  20. Winkler T, Jahn S, Kalden JR (1991) IgG human monoclonal anti-DNA autoantibodies from patients with systemic lupus crythematosus. Clin Exp Immunol 85:379–385

    PubMed  Google Scholar 

  21. Hiepe F, Kiessig ST, Jahn S, Volk HD, Grunow R, Apostoloff E, Bachr R von (1986) A sensitive and class-specific solid phase enzyme immunoassay for anti-DNA autoantibodies in supernatants of lymphocyte cultures and human hybridomas. Biomed Biochem Acta 1986; 45:1229–1235

    Google Scholar 

  22. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor Laboratory Press

  23. Ravetch JV, Siebenlist U, Korsmayer S, Waldmann T, Leder P (1981) Structure of the human immunoglobulin μ locus: characterization of embryonic and rearranged J and D genes. Cell 27:583–591

    Article  PubMed  Google Scholar 

  24. Sanger F, Nicklen S, Coulson A (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    PubMed  Google Scholar 

  25. Marks JD, Tristem M, Karpas A, Winter G (1991) Oligo nucleotide primers for polymerase chain reaction amplification of human Ig variable genes and design of family-specific oligonucleotide probes. Eur J Immunol. 21:985–991

    PubMed  Google Scholar 

  26. Larrick JW, Danielsson L, Brenner CA, Abrahamson M, Frey KE, Borrebaek CAK (1989) Rapid cloning of rearranged immunoglobulin genes from human hybridoma cells using mixed primers and the polymerase chain reaction. Biochem Biophys Res Comm 160:1250–1256

    PubMed  Google Scholar 

  27. Songsivilai S, Bye JM, Marks JD, Hughes-Jones NC (1990) Cloning and sequencing of human lambda immunoglobulin genes by the polymerase chain reaction. Eur J Immunol 20:2661–2666

    PubMed  Google Scholar 

  28. Berman JE, Mellis SJ, Pollock R, Smith CC, Suh HS, Heinke B, Kowall C, Surti U, Chess L, Cantor CR, Alt CR (1988) Content and organization of the human IgVH locus: definition of three new VH families and linkage to the IgCH locus. EMBO J 7:727–738

    PubMed  Google Scholar 

  29. Ternynck T, Avrameas S (1986) Murine natural monoclonal antibodies: a study of their polyspecificities and their affinity. Immunol Rev 94:99–112

    PubMed  Google Scholar 

  30. Hampe J, Nürnberg P, Epplen C, Jahn S, Grunow R, Epplen JT (1992) Oligonucleotide fingerprinting as a means to identify and survey long-term cultured B cell hybridomas and T cell lines. Hum Antibodies Hybridomas 3:186–190

    PubMed  Google Scholar 

  31. Ichihara Y, Matsuoka H, Kurosawa Y (1988) Organization of human immunoglobulin heavy chain diversity gene locus. EMBO J 7:4141–4150

    PubMed  Google Scholar 

  32. Siebenlist U, Ravetch JW, Korsmeyer S, Waldman T, Leder L (1981) Human immunoglobulin D segment encoded in tandem multigenic families. Nature 294:631–635

    PubMed  Google Scholar 

  33. Yamasaki N, Komori S, Watanabe T (1987) Complementary DNA for a human subgroup IV immunoglobulin lambdachain. Mol Immunol 24:981–984

    Article  PubMed  Google Scholar 

  34. Harindranath N, Goldfarb IS, Ikematsu H, Burastero SE, Wilder RL, Notkins AL, Casali P (1991) Complete sequence of the genes encoding the VH and the VL regions of low and high affinity monoclonal IgM and IgA1 rheumatoid factors produced by CD5+ B cells from a patient with rheumatoid arthritis. Int J Immunol 3:865–875

    Google Scholar 

  35. Chuchana P, Blancer A, Brockly F, Alexandre D, Lefranc G, Lefranc MP (1990) Definition of the human immunoglobulin variable lambda (IGLV) gene subgroups. Eur J Immunol 20:1317–1325

    PubMed  Google Scholar 

  36. Meng QH, Rauch J (1990) Differences between human hybridoma platelet-binding antibodies derived from systemic lupus erythematosus patients and normal individuals. Autoimmunity 5:151–167

    PubMed  Google Scholar 

  37. Tron F, Bach JF (1989) Molecular and genetic characterization of pathogenic autoantibodies. J Autoimmun 2:311–320

    Article  PubMed  Google Scholar 

  38. Andrezejewski C, Rauch J, Lafer E, Stollar BD, Schwartz RS (1981) Antigen-binding diversity and idiotypic cross-reaction among hybridoma autoantibodies to DNA. J Immunol 126:226–231

    PubMed  Google Scholar 

  39. Campbell MJ, Zelenetz AD, Levy S, Levy R (1992) Use of family specific leader region primers for PCR amplification of the human heavy chain variable region gene repertoire. Mol Immunol 29:193–203

    Article  PubMed  Google Scholar 

  40. Schutte MEM, Ebeling SB, Akkermans KE, Gmelig-Meyling FJH, Logtenberg T (1991) Antibody specificity and immunoglobulin VH gene utilization of human monoclonal CD5+ B cell lines. Eur J Immunol 21:1115–1121

    PubMed  Google Scholar 

  41. Olee T, Lu EW, Huang DF, Soto-Gil R, Deftos M, Kozin F, Carson DA, Chen PP (1992) Genetic analysis of self-associating immunoglobulin G rheumatoid factors from two rheumatoid synovia implicates an antigen-driven response. J Exp Med 175:831–842

    Article  PubMed  Google Scholar 

  42. Kipps TJ, Tomhave E, Pratt CF, Duffy S, Chen PP, Carson D (1989) Developmentally restricted immunoglobulin heavy chain variable region gene expressed at high frequency in chronic lymphocytic leukemia. Proc Natl Acad Sci 86:5913–5917

    PubMed  Google Scholar 

  43. Deanne M, Norton JD (1990) Immunoglobulin heavy chain variable region family is independent of tumor cell phenotype in human B lineage leukemias. Eur J Immunol 20:2209–2217

    PubMed  Google Scholar 

  44. Schroeder HW, Hillson JL, Perlmutter RM (1987) Early restriction of the human antibody repertoire. Science 238:791–793

    PubMed  Google Scholar 

  45. Walter MA, Dosch HM, Cox DW (1991) A deletion map of the human immunoglobulin heavy chain variable region. J Exp Med 174:335–349

    Article  PubMed  Google Scholar 

  46. Shin EK, Matsuda F, Nagaoka H, Fukita Y, Imai T, Yokoyama K, Soeda E, Honjo T (1991) Physical map of the 3′ region of the human immunoglobulin heavy chain locus: clustering of autoantibody-related variable segments in one haplotype. EMBO J 10:3641–3645

    PubMed  Google Scholar 

  47. Nickerson K, Berman J, Glickman E, Chess L, Alt FW (1989) Early human IgH gene assembly in Epstein-Barr virus transformed fetal B cell lines. J Exp Med 169:1391–1403

    Article  PubMed  Google Scholar 

  48. Yancopoulos GD, Desiderio SV, Paskind M, Kearney JF, Baltimore D (1984) Preferential utilization of the most JH proximal VH segments in pre-B cell lines. Nature 311:727–733

    PubMed  Google Scholar 

  49. Shlomchik M, Mascelli M, Shan H, Radic MZ, Pisetsky D, Marshak-Rothstein A, Weigert M (1990) Anti-DNA antibodies from autoimmune mice arise by clonal expansion and somatic mutation. J Exp Med 171:265–297

    Article  PubMed  Google Scholar 

  50. Sanz I (1991) Multiple mechanisms participate in the generation of diversity of human H chain CDR3 regions. J Immunol 147:1720–1729

    PubMed  Google Scholar 

  51. Wysocki L, Manser T, Gefter R (1986) Somatic evolution of variable structures during an immune response. Proc Natl Acad Sci USA 83:1847–1854

    PubMed  Google Scholar 

  52. Schroeder HW, Wang JY (1990) Preferential utilization of conserved immunoglobulin heavy chain variable gene segments during human fetal life. Proc Natl Acad Sci 87:6146–6150

    PubMed  Google Scholar 

  53. Davidson A, Manheimer-Lory A, Aranow C, Peterson R, Hannigan N, Diamond B (1990) Molecular characterization of a somatically mutated anti-DNA antibody bearing two systemic lupus erythematosus-related idiotypes. J Clin Invest 85:1401–1409

    PubMed  Google Scholar 

  54. Borghesi-Nicoletti C, Schulze DH (1991) Polymerase chain reaction of genes flanked by short noncontiguous sequence motifs. Anal Biochem 1991; 192:449–452

    Google Scholar 

  55. Tomlinson IM, Walter G, Marks JD, Llewelyn MB, Winter G (1992) The repertoire of human germline VH sequences. Reveals about fifty groups of VH sequences with different hypervariable loops. J Mol Biol 227:776–798

    PubMed  Google Scholar 

  56. Matsuda F, Shin EK, Hirabayashi Y, Nagaoka H, Yoshida MC, Shu QZ, Honjo T (1990) Organization of variable region segments of the human immunoglobulin heavy chain: duplication of the D5 cluster within the locus and interchromosomal translocation of variable region segments. EMBO J 9:2501–2506

    PubMed  Google Scholar 

  57. Seeman NC, Rosenberg JM, Rich R (1976) Sequence-specific recognition of double-helical nucleic acids by protein. Proc Natl Acad Sci USA 73:804–809

    PubMed  Google Scholar 

  58. McClarin JA, Frederick CA, Wang B, Greene P, Boyer HW, Grable J, Rosenberg JM (1986) Structure of the DNA-EcoRI endonuclease recognition complex at 3 Å resolution. Science 234:1526–1528

    PubMed  Google Scholar 

  59. Herron JN, He XM, Ballard DW, Blier PR, Pace PE, Bothwell ALM, Voss EW, Edmundson EB (1991) An autoantibody to single-stranded DNA: comparison of the three-dimensional structures of the unligated Fab and a deoxynucleotide-Fab complex. Proteins 11:159–175

    PubMed  Google Scholar 

  60. Berek C, Milstein C (1987) Mutational drift and repertoire shift in the maturation of the immune response. Immunol Rev 96:23–41

    PubMed  Google Scholar 

  61. Rajewsky K, Förster I, Cumano A (1987) Evolutionary and somatic selection of the antibody repertoire in the mouse. Science 238:1088–1094

    PubMed  Google Scholar 

  62. Winkler T, Fehr H, Kalden JR (1992) Analysis of immunoglobulin variable region genes from human IgG anti-dsDNA hybridomas. Eur J Immunol 22:1719–1728

    PubMed  Google Scholar 

  63. Es JH van, Gmelig-Meyling FH, Akker WR van de, Aanstoot H, Derksen RH, Logtenberg T (1991) Somatic mutations in the variable regions of a human IgG double-stranded DNA auto-antibody suggest a role for antigen in the induction of systemic lupus erythematosus. J Exp Med 173:461–470

    Article  PubMed  Google Scholar 

  64. Pascual V, Victor K, Lelsz D, Spellerberg MB, Hamblin TJ, Thompson KM, Randen I, Natvig J, Capra JD, Stevenson FK (1991) Nucleotide sequence analysis of the V regions of two IgM cold agglutinins. Evidence that the VH4-21 gene segment is responsible for the major cross-reactive idiotype J Immunol 146:4385–4391

    PubMed  Google Scholar 

  65. Radbruch A, ZaissS, Kappen C, Brüggemann M, Beyreuther M, Rajewsky K (1985) Drastic change in idiotypic but not antigen-binding specificity of an antibody by a single amino acid substitution. Nature 315:506–509

    PubMed  Google Scholar 

  66. Reidl LS, Friedman DF, Goldman J, Hardy RR, Jefferies LC, Silberstein LE (1991) Structural basis of a conserved idiotype expressed by an autoreactive murine B cell lymphoma. J Immunol 1991; 147:3029–3031

    Google Scholar 

  67. Taki S, Hirose S, Kinoshita K, Nishimura H, Shimamura T, Hamuro J, Shirai T (1992) Somatically mutated IgG anti-DNA antibody related to germ-line encoded IgM anti-DNA antibody. Eur J Immunol 22:987–995

    PubMed  Google Scholar 

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  1. T. Winkler

    Present address: Basel Institute for Immunology, Grenzacher Strasse 487, CH-4058, Basel, Switzerland

Authors and Affiliations

  1. Institute for Medical Immunology, Charité, Humboldt University, Berlin, Germany

    S. Jahn, B. Niemann & R. von Baehr

  2. Institute for Clinical Immunology and Rheumatology, University of Erlangen, Germany

    T. Winkler & J. R. Kalden

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  1. S. Jahn
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  2. B. Niemann
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  3. T. Winkler
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  4. J. R. Kalden
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Jahn, S., Niemann, B., Winkler, T. et al. Expansion of a B-lymphocyte clone producing IgM auto-antibodies encoded by a somatically mutated VHI gene in the spleen of an autoimmune patient. Rheumatol Int 13, 187–196 (1994). https://doi.org/10.1007/BF00390266

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  • DOI: https://doi.org/10.1007/BF00390266

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