Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1991 Feb 1;173(2):461–470. doi: 10.1084/jem.173.2.461

Somatic mutations in the variable regions of a human IgG anti-double- stranded DNA autoantibody suggest a role for antigen in the induction of systemic lupus erythematosus

PMCID: PMC2118793  PMID: 1899104

Abstract

The processes that govern the generation of pathogenic anti-DNA autoantibodies in human systemic lupus erythematosus (SLE) are largely unknown. Autoantibodies may arise as a consequence of polyclonal B cell activation and/or antigen-driven B cell activation and selection. The role of these processes in humoral autoimmunity may be studied by molecular genetic analysis of immunoglobulin (Ig) variable (V) regions of antibodies that are characteristic of SLE. We have analyzed the gene elements that encode a high affinity, IgG anti-double-stranded DNA autoantibody secreted by a monoclonal Epstein-Barr virus (EBV)- transformed cell line derived from a patient with active SLE. In addition, we have identified, cloned, and sequenced the germline counterparts of the VH and VL genes expressed in this autoantibody. The comparison of both sets of gene elements shows that the autoantibody VH and VL regions harbor numerous somatic mutations characteristic of an antigen-driven immune response. The light chain expressed in this autoantibody is a somatically mutated variant of the kv325 germline gene that is frequently associated with paraproteins having autoantibody activity and with Ig molecules produced by malignant B cells that express the CD5 antigen. Furthermore, the utilized DH segment has been repeatedly found in multireactive, low affinity IgM anti-DNA autoantibodies from SLE patients and healthy individuals. These results suggest that pathogenic IgG anti-DNA autoantibodies in human SLE may arise through antigen-driven selection of somatic mutations in the gene elements that frequently encode multireactive IgM autoantibodies.

Full Text

The Full Text of this article is available as a PDF (955.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berman J. E., Mellis S. J., Pollock R., Smith C. L., Suh H., Heinke B., Kowal C., Surti U., Chess L., Cantor C. R. Content and organization of the human Ig VH locus: definition of three new VH families and linkage to the Ig CH locus. EMBO J. 1988 Mar;7(3):727–738. doi: 10.1002/j.1460-2075.1988.tb02869.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cairns E., Block J., Bell D. A. Anti-DNA autoantibody-producing hybridomas of normal human lymphoid cell origin. J Clin Invest. 1984 Sep;74(3):880–887. doi: 10.1172/JCI111505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cairns E., Kwong P. C., Misener V., Ip P., Bell D. A., Siminovitch K. A. Analysis of variable region genes encoding a human anti-DNA antibody of normal origin. Implications for the molecular basis of human autoimmune responses. J Immunol. 1989 Jul 15;143(2):685–691. [PubMed] [Google Scholar]
  4. Chen P. P., Albrandt K., Kipps T. J., Radoux V., Liu F. T., Carson D. A. Isolation and characterization of human VkIII germ-line genes. Implications for the molecular basis of human VkIII light chain diversity. J Immunol. 1987 Sep 1;139(5):1727–1733. [PubMed] [Google Scholar]
  5. Davidson A., Smith A., Katz J., Preud'Homme J. L., Solomon A., Diamond B. A cross-reactive idiotype on anti-DNA antibodies defines a H chain determinant present almost exclusively on IgG antibodies. J Immunol. 1989 Jul 1;143(1):174–180. [PubMed] [Google Scholar]
  6. Dersimonian H., McAdam K. P., Mackworth-Young C., Stollar B. D. The recurrent expression of variable region segments in human IgM anti-DNA autoantibodies. J Immunol. 1989 Jun 1;142(11):4027–4033. [PubMed] [Google Scholar]
  7. Dziarski R. Autoimmunity: polyclonal activation or antigen induction? Immunol Today. 1988 Nov;9(11):340–342. doi: 10.1016/0167-5699(88)91333-3. [DOI] [PubMed] [Google Scholar]
  8. Dziarski R. Preferential induction of autoantibody secretion in polyclonal activation by peptidoglycan and lipopolysaccharide. I. In vitro studies. J Immunol. 1982 Mar;128(3):1018–1025. [PubMed] [Google Scholar]
  9. Ebling F., Hahn B. H. Restricted subpopulations of DNA antibodies in kidneys of mice with systemic lupus. Comparison of antibodies in serum and renal eluates. Arthritis Rheum. 1980 Apr;23(4):392–403. doi: 10.1002/art.1780230402. [DOI] [PubMed] [Google Scholar]
  10. Eilat D., Webster D. M., Rees A. R. V region sequences of anti-DNA and anti-RNA autoantibodies from NZB/NZW F1 mice. J Immunol. 1988 Sep 1;141(5):1745–1753. [PubMed] [Google Scholar]
  11. Gavalchin J., Datta S. K. The NZB X SWR model of lupus nephritis. II. Autoantibodies deposited in renal lesions show a distinctive and restricted idiotypic diversity. J Immunol. 1987 Jan 1;138(1):138–148. [PubMed] [Google Scholar]
  12. Goñi F. R., Chen P. P., McGinnis D., Arjonilla M. L., Fernandez J., Carson D., Solomon A., Mendez E., Frangione B. Structural and idiotypic characterization of the L chains of human IgM autoantibodies with different specificities. J Immunol. 1989 May 1;142(9):3158–3163. [PubMed] [Google Scholar]
  13. Hoch S., Schwaber J. Identification and sequence of the VH gene elements encoding a human anti-DNA antibody. J Immunol. 1987 Sep 1;139(5):1689–1693. [PubMed] [Google Scholar]
  14. Ichihara Y., Matsuoka H., Kurosawa Y. Organization of human immunoglobulin heavy chain diversity gene loci. EMBO J. 1988 Dec 20;7(13):4141–4150. doi: 10.1002/j.1460-2075.1988.tb03309.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kipps T. J., Fong S., Tomhave E., Chen P. P., Goldfien R. D., Carson D. A. High-frequency expression of a conserved kappa light-chain variable-region gene in chronic lymphocytic leukemia. Proc Natl Acad Sci U S A. 1987 May;84(9):2916–2920. doi: 10.1073/pnas.84.9.2916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kipps T. J., Robbins B. A., Carson D. A. Uniform high frequency expression of autoantibody-associated crossreactive idiotypes in the primary B cell follicles of human fetal spleen. J Exp Med. 1990 Jan 1;171(1):189–196. doi: 10.1084/jem.171.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klinman D. M., Steinberg A. D. Systemic autoimmune disease arises from polyclonal B cell activation. J Exp Med. 1987 Jun 1;165(6):1755–1760. doi: 10.1084/jem.165.6.1755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kocks C., Rajewsky K. Stable expression and somatic hypermutation of antibody V regions in B-cell developmental pathways. Annu Rev Immunol. 1989;7:537–559. doi: 10.1146/annurev.iy.07.040189.002541. [DOI] [PubMed] [Google Scholar]
  19. Lee K. H., Matsuda F., Kinashi T., Kodaira M., Honjo T. A novel family of variable region genes of the human immunoglobulin heavy chain. J Mol Biol. 1987 Jun 20;195(4):761–768. doi: 10.1016/0022-2836(87)90482-7. [DOI] [PubMed] [Google Scholar]
  20. Logtenberg T., Kroon A., Gmelig-Meyling F. H., Ballieux R. E. Analysis of the human tonsil B cell repertoire by somatic hybridization: occurrence of both "monospecific" and "multispecific" (auto)antibody-secreting cells. Eur J Immunol. 1987 Jun;17(6):855–859. doi: 10.1002/eji.1830170619. [DOI] [PubMed] [Google Scholar]
  21. Logtenberg T., Young F. M., Van Es J. H., Gmelig-Meyling F. H., Alt F. W. Autoantibodies encoded by the most Jh-proximal human immunoglobulin heavy chain variable region gene. J Exp Med. 1989 Oct 1;170(4):1347–1355. doi: 10.1084/jem.170.4.1347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Logtenberg T., Young F. M., van Es J., Gmelig-Meyling F. H., Berman J. E., Alt F. W. Frequency of VH-gene utilization in human EBV-transformed B-cell lines: the most JH-proximal VH segment encodes autoantibodies. J Autoimmun. 1989 Jun;2 (Suppl):203–213. doi: 10.1016/0896-8411(89)90132-7. [DOI] [PubMed] [Google Scholar]
  23. Mudde G. C., Verberne C. J., Groeneveld K., De Gast G. C. Human tonsil B lymphocyte function. I. The proliferative response to Staphylococcus aureus and pokeweed mitogen in relation to surface heavy chains mu and delta. Clin Exp Immunol. 1984 Jun;56(3):709–715. [PMC free article] [PubMed] [Google Scholar]
  24. Poncet P., Matthes T., Billecocq A., Dighiero G. Immunochemical studies of polyspecific natural autoantibodies: charge, lipid reactivity, Fab'2 fragments activity and complement fixation. Mol Immunol. 1988 Oct;25(10):981–989. doi: 10.1016/0161-5890(88)90004-1. [DOI] [PubMed] [Google Scholar]
  25. ROSENTHAL M. C., PISCIOTTA A. V., KOMNINOS Z. D., GOLDENBERG H., DAMESHEK W. The auto-immune hemolytic anemia of malignant lymphocytic disease. Blood. 1955 Mar;10(3):197–227. [PubMed] [Google Scholar]
  26. Radoux V., Chen P. P., Sorge J. A., Carson D. A. A conserved human germline V kappa gene directly encodes rheumatoid factor light chains. J Exp Med. 1986 Dec 1;164(6):2119–2124. doi: 10.1084/jem.164.6.2119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanz I., Kelly P., Williams C., Scholl S., Tucker P., Capra J. D. The smaller human VH gene families display remarkably little polymorphism. EMBO J. 1989 Dec 1;8(12):3741–3748. doi: 10.1002/j.1460-2075.1989.tb08550.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Saxon A., Feldhaus J., Robins R. A. Single step separation of human T and B cells using AET treated srbc rosettes. J Immunol Methods. 1976;12(3-4):285–288. doi: 10.1016/0022-1759(76)90050-8. [DOI] [PubMed] [Google Scholar]
  29. Schroeder H. W., Jr, Hillson J. L., Perlmutter R. M. Structure and evolution of mammalian VH families. Int Immunol. 1990;2(1):41–50. doi: 10.1093/intimm/2.1.41. [DOI] [PubMed] [Google Scholar]
  30. Seeman N. C., Rosenberg J. M., Rich A. Sequence-specific recognition of double helical nucleic acids by proteins. Proc Natl Acad Sci U S A. 1976 Mar;73(3):804–808. doi: 10.1073/pnas.73.3.804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shlomchik M. J., Aucoin A. H., Pisetsky D. S., Weigert M. G. Structure and function of anti-DNA autoantibodies derived from a single autoimmune mouse. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9150–9154. doi: 10.1073/pnas.84.24.9150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shlomchik M. J., Marshak-Rothstein A., Wolfowicz C. B., Rothstein T. L., Weigert M. G. The role of clonal selection and somatic mutation in autoimmunity. 1987 Aug 27-Sep 2Nature. 328(6133):805–811. doi: 10.1038/328805a0. [DOI] [PubMed] [Google Scholar]
  33. Shlomchik M., Mascelli M., Shan H., Radic M. Z., Pisetsky D., Marshak-Rothstein A., Weigert M. Anti-DNA antibodies from autoimmune mice arise by clonal expansion and somatic mutation. J Exp Med. 1990 Jan 1;171(1):265–292. doi: 10.1084/jem.171.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shlomchik M., Nemazee D., van Snick J., Weigert M. Variable region sequences of murine IgM anti-IgG monoclonal autoantibodies (rheumatoid factors). II. Comparison of hybridomas derived by lipopolysaccharide stimulation and secondary protein immunization. J Exp Med. 1987 Apr 1;165(4):970–987. doi: 10.1084/jem.165.4.970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Shoenfeld Y., Isenberg D. A., Rauch J., Madaio M. P., Stollar B. D., Schwartz R. S. Idiotypic cross-reactions of monoclonal human lupus autoantibodies. J Exp Med. 1983 Sep 1;158(3):718–730. doi: 10.1084/jem.158.3.718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Smeenk R. J., Brinkman K., van den Brink H. G., Westgeest A. A. Reaction patterns of monoclonal antibodies to DNA. J Immunol. 1988 Jun 1;140(11):3786–3792. [PubMed] [Google Scholar]
  37. Sontheimer R. D., Gilliam J. N. DNA antibody class, subclass, and complement fixation in systemic lupus erythematosus with and without nephritis. Clin Immunol Immunopathol. 1978 Aug;10(4):459–467. doi: 10.1016/0090-1229(78)90158-7. [DOI] [PubMed] [Google Scholar]
  38. Sthoeger Z. M., Wakai M., Tse D. B., Vinciguerra V. P., Allen S. L., Budman D. R., Lichtman S. M., Schulman P., Weiselberg L. R., Chiorazzi N. Production of autoantibodies by CD5-expressing B lymphocytes from patients with chronic lymphocytic leukemia. J Exp Med. 1989 Jan 1;169(1):255–268. doi: 10.1084/jem.169.1.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Swaak A. J., Aarden L. A., Statius van Eps L. W., Feltkamp T. E. Anti-dsDNA and complement profiles as prognostic guides in systemic lupus erythematosus. Arthritis Rheum. 1979 Mar;22(3):226–235. doi: 10.1002/art.1780220304. [DOI] [PubMed] [Google Scholar]
  40. Tan E. M. Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol. 1989;44:93–151. doi: 10.1016/s0065-2776(08)60641-0. [DOI] [PubMed] [Google Scholar]
  41. Winfield J. B., Faiferman I., Koffler D. Avidity of anti-DNA antibodies in serum and IgG glomerular eluates from patients with systemic lupus erythematosus. Association of high avidity antinative DNA antibody with glomerulonephritis. J Clin Invest. 1977 Jan;59(1):90–96. doi: 10.1172/JCI108626. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES