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. 1996 Oct 15;98(8):1843–1850. doi: 10.1172/JCI118985

Characterization of a germline Vk gene encoding cationic anti-DNA antibody and role of receptor editing for development of the autoantibody in patients with systemic lupus erythematosus.

N Suzuki 1, T Harada 1, S Mihara 1, T Sakane 1
PMCID: PMC507624  PMID: 8878436

Abstract

We found previously that cationic anti-DNA autoantibodies (autoAbs) have nephritogenic potential and usage of a specific germline Vk gene, A30, has major influences on cationic charge of the autoAb in human lupus nephritis. In the present study, we have characterized A30 germline Vk gene using cosmid cloning technique in patients with SLE. A30 gene locus locates in less than 250 kb from the Ck region, and the cationic anti-DNA mRNA used the upstream Jk2 gene, indicating that cationic anti-DNA mRNA is a product of primary gene rearrangement. By using PCR technique, we found that A30 gene locus in the genome was defective in eight out of nine SLE patients without nephritis. In contrast, all nine patients with lupus nephritis had intact A30 gene. The presence and absence of A30 gene was associated with the development of lupus nephritis or not (P < 0.01, by Fisher's exact test, two-sided). It was thus suggested that absence of functional A30 gene may rescue from developing lupus nephritis in the patients. A30 is reported to be a potentially functional but rarely expressed Vk gene in humans. It is possible that normal B cells edit primarily rearranged A30 gene with autoreactive potentials by receptor editing mechanism for changing the affinity of the B cell Ag receptor to avoid self-reactivity, whereas SLE B cells may have a defect in this mechanism. Indeed, we found that normal B cells edit A30-Jk2 gene in their genome possibly by inversion mechanism, whereas SLE B cells contain rearranged A30-Jk2-Ck gene in the genome and express A30-associated mRNA, suggesting that receptor editing mechanism is also defective in patients with SLE. Our study suggests that polymorphism of Ig Vk locus, and failure of receptor editing may contribute to the development of pathogenic anti-DNA responses in humans.

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Selected References

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  1. Barnes W. M. PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2216–2220. doi: 10.1073/pnas.91.6.2216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bensimon C., Chastagner P., Zouali M. Human lupus anti-DNA autoantibodies undergo essentially primary V kappa gene rearrangements. EMBO J. 1994 Jul 1;13(13):2951–2962. doi: 10.1002/j.1460-2075.1994.tb06593.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chastagner P., Demaison C., Thèze J., Zouali M. Clonotypic dominance and variable gene elements of pathogenic anti-DNA autoantibodies from a single patient with lupus. Scand J Immunol. 1994 Feb;39(2):165–178. doi: 10.1111/j.1365-3083.1994.tb03356.x. [DOI] [PubMed] [Google Scholar]
  4. Chen C., Nagy Z., Radic M. Z., Hardy R. R., Huszar D., Camper S. A., Weigert M. The site and stage of anti-DNA B-cell deletion. Nature. 1995 Jan 19;373(6511):252–255. doi: 10.1038/373252a0. [DOI] [PubMed] [Google Scholar]
  5. Chen P. P., Siminovitch K. A., Olsen N. J., Erger R. A., Carson D. A. A highly informative probe for two polymorphic Vh gene regions that contain one or more autoantibody-associated Vh genes. J Clin Invest. 1989 Aug;84(2):706–710. doi: 10.1172/JCI114218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cheng S., Chang S. Y., Gravitt P., Respess R. Long PCR. Nature. 1994 Jun 23;369(6482):684–685. doi: 10.1038/369684a0. [DOI] [PubMed] [Google Scholar]
  7. Demaison C., Chastagner P., Thèze J., Zouali M. Somatic diversification in the heavy chain variable region genes expressed by human autoantibodies bearing a lupus-associated nephritogenic anti-DNA idiotype. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):514–518. doi: 10.1073/pnas.91.2.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Diamond B., Katz J. B., Paul E., Aranow C., Lustgarten D., Scharff M. D. The role of somatic mutation in the pathogenic anti-DNA response. Annu Rev Immunol. 1992;10:731–757. doi: 10.1146/annurev.iy.10.040192.003503. [DOI] [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. Erikson J., Radic M. Z., Camper S. A., Hardy R. R., Carmack C., Weigert M. Expression of anti-DNA immunoglobulin transgenes in non-autoimmune mice. Nature. 1991 Jan 24;349(6307):331–334. doi: 10.1038/349331a0. [DOI] [PubMed] [Google Scholar]
  11. Evans G. A., Wahl G. M. Cosmid vectors for genomic walking and rapid restriction mapping. Methods Enzymol. 1987;152:604–610. doi: 10.1016/0076-6879(87)52068-7. [DOI] [PubMed] [Google Scholar]
  12. Gay D., Saunders T., Camper S., Weigert M. Receptor editing: an approach by autoreactive B cells to escape tolerance. J Exp Med. 1993 Apr 1;177(4):999–1008. doi: 10.1084/jem.177.4.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goodnow C. C., Crosbie J., Adelstein S., Lavoie T. B., Smith-Gill S. J., Brink R. A., Pritchard-Briscoe H., Wotherspoon J. S., Loblay R. H., Raphael K. Altered immunoglobulin expression and functional silencing of self-reactive B lymphocytes in transgenic mice. Nature. 1988 Aug 25;334(6184):676–682. doi: 10.1038/334676a0. [DOI] [PubMed] [Google Scholar]
  14. Harada T., Suzuki N., Mizushima Y., Sakane T. Usage of a novel class of germ-line Ig variable region gene for cationic anti-DNA autoantibodies in human lupus nephritis and its role for the development of the disease. J Immunol. 1994 Nov 15;153(10):4806–4815. [PubMed] [Google Scholar]
  15. Jang Y. J., Stollar B. D. Ultraviolet cross-linking of helical oligonucleotides to two monoclonal MRL-1pr/1pr anti-DNA autoantibodies. Variations in H and L chain binding to DNA. J Immunol. 1990 Nov 15;145(10):3353–3359. [PubMed] [Google Scholar]
  16. Kariya Y., Kato K., Hayashizaki Y., Himeno S., Tarui S., Matsubara K. Expression of human gastrin gene in normal and gastrinoma tissues. Gene. 1986;50(1-3):345–352. doi: 10.1016/0378-1119(86)90338-0. [DOI] [PubMed] [Google Scholar]
  17. Klein R., Jaenichen R., Zachau H. G. Expressed human immunoglobulin kappa genes and their hypermutation. Eur J Immunol. 1993 Dec;23(12):3248–3262. doi: 10.1002/eji.1830231231. [DOI] [PubMed] [Google Scholar]
  18. Lautner-Rieske A., Huber C., Meindl A., Pargent W., Schäble K. F., Thiebe R., Zocher I., Zachau H. G. The human immunoglobulin kappa locus. Characterization of the duplicated A regions. Eur J Immunol. 1992 Apr;22(4):1023–1029. doi: 10.1002/eji.1830220422. [DOI] [PubMed] [Google Scholar]
  19. Mahmoudi M., Denomme G. A., Edwards J. Y., Bell D. A., Cairns E. The role of the immunoglobulin heavy chain in human anti-DNA antibody binding specificity. Arthritis Rheum. 1995 Mar;38(3):389–395. doi: 10.1002/art.1780380316. [DOI] [PubMed] [Google Scholar]
  20. Malcolm S., Barton P., Murphy C., Ferguson-Smith M. A., Bentley D. L., Rabbitts T. H. Localization of human immunoglobulin kappa light chain variable region genes to the short arm of chromosome 2 by in situ hybridization. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4957–4961. doi: 10.1073/pnas.79.16.4957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Meindl A., Kellner H., Schattenkirchner M., Zachau H. G. Polymorphisms in the VK gene patterns of rheumatoid arthritis patients and control individuals. Exp Clin Immunogenet. 1990;7(1):20–25. [PubMed] [Google Scholar]
  22. Meindl A., Klobeck H. G., Ohnheiser R., Zachau H. G. The V kappa gene repertoire in the human germ line. Eur J Immunol. 1990 Aug;20(8):1855–1863. doi: 10.1002/eji.1830200834. [DOI] [PubMed] [Google Scholar]
  23. Mochizuki M., Suzuki N., Takeno M., Nagafuchi H., Harada T., Kaneoka H., Yamashita N., Hirayama K., Nakajima T., Mizushima Y. Fine antigen specificity of human gamma delta T cell lines (V gamma 9+) established by repetitive stimulation with a serotype (KTH-1) of a gram-positive bacterium, Streptococcus sanguis. Eur J Immunol. 1994 Jul;24(7):1536–1543. doi: 10.1002/eji.1830240712. [DOI] [PubMed] [Google Scholar]
  24. Moxley G. Variable-constant segment genotype of immunoglobulin kappa is associated with increased risk for rheumatoid arthritis. Arthritis Rheum. 1992 Jan;35(1):19–25. doi: 10.1002/art.1780350104. [DOI] [PubMed] [Google Scholar]
  25. Muryoi T., Sasaki T., Hatakeyama A., Shibata S., Suzuki M., Seino J., Yoshinaga K. Clonotypes of anti-DNA antibodies expressing specific idiotypes in immune complexes of patients with active lupus nephritis. J Immunol. 1990 May 15;144(10):3856–3861. [PubMed] [Google Scholar]
  26. Nemazee D. A., Bürki K. Clonal deletion of B lymphocytes in a transgenic mouse bearing anti-MHC class I antibody genes. Nature. 1989 Feb 9;337(6207):562–566. doi: 10.1038/337562a0. [DOI] [PubMed] [Google Scholar]
  27. Olee T., Yang P. M., Siminovitch K. A., Olsen N. J., Hillson J., Wu J., Kozin F., Carson D. A., Chen P. P. Molecular basis of an autoantibody-associated restriction fragment length polymorphism that confers susceptibility to autoimmune diseases. J Clin Invest. 1991 Jul;88(1):193–203. doi: 10.1172/JCI115277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pargent W., Schäble K. F., Zachau H. G. Polymorphisms and haplotypes in the human immunoglobulin kappa locus. Eur J Immunol. 1991 Aug;21(8):1829–1835. doi: 10.1002/eji.1830210808. [DOI] [PubMed] [Google Scholar]
  29. Radic M. Z., Erikson J., Litwin S., Weigert M. B lymphocytes may escape tolerance by revising their antigen receptors. J Exp Med. 1993 Apr 1;177(4):1165–1173. doi: 10.1084/jem.177.4.1165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Radic M. Z., Mascelli M. A., Erikson J., Shan H., Weigert M. Ig H and L chain contributions to autoimmune specificities. J Immunol. 1991 Jan 1;146(1):176–182. [PubMed] [Google Scholar]
  31. Radic M. Z., Weigert M. Genetic and structural evidence for antigen selection of anti-DNA antibodies. Annu Rev Immunol. 1994;12:487–520. doi: 10.1146/annurev.iy.12.040194.002415. [DOI] [PubMed] [Google Scholar]
  32. Scott M. G., Crimmins D. L., McCourt D. W., Chung G., Schäble K. F., Thiebe R., Quenzel E. M., Zachau H. G., Nahm M. H. Clonal characterization of the human IgG antibody repertoire to Haemophilus influenzae type b polysaccharide. IV. The less frequently expressed VL are heterogeneous. J Immunol. 1991 Dec 1;147(11):4007–4013. [PubMed] [Google Scholar]
  33. Shin E. K., Matsuda F., Nagaoka H., Fukita Y., Imai T., Yokoyama K., Soeda E., Honjo T. Physical map of the 3' region of the human immunoglobulin heavy chain locus: clustering of autoantibody-related variable segments in one haplotype. EMBO J. 1991 Dec;10(12):3641–3645. doi: 10.1002/j.1460-2075.1991.tb04930.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Skowronski J., Fanning T. G., Singer M. F. Unit-length line-1 transcripts in human teratocarcinoma cells. Mol Cell Biol. 1988 Apr;8(4):1385–1397. doi: 10.1128/mcb.8.4.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Smith H. R., Steinberg A. D. Autoimmunity--a perspective. Annu Rev Immunol. 1983;1:175–210. doi: 10.1146/annurev.iy.01.040183.001135. [DOI] [PubMed] [Google Scholar]
  36. Suzuki N., Harada T., Mizushima Y., Sakane T. Possible pathogenic role of cationic anti-DNA autoantibodies in the development of nephritis in patients with systemic lupus erythematosus. J Immunol. 1993 Jul 15;151(2):1128–1136. [PubMed] [Google Scholar]
  37. Suzuki N., Otuka I., Harada T., Mizushima Y., Sakane T. Preferential adsorption of cationic anti-DNA antibodies with immobilized polyanionic compounds, dextran sulfate. Autoimmunity. 1994;19(2):105–112. doi: 10.3109/08916939409009538. [DOI] [PubMed] [Google Scholar]
  38. Suzuki N., Sakane T., Engleman E. G. Anti-DNA antibody production by CD5+ and CD5- B cells of patients with systemic lupus erythematosus. J Clin Invest. 1990 Jan;85(1):238–247. doi: 10.1172/JCI114418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tan E. M., Cohen A. S., Fries J. F., Masi A. T., McShane D. J., Rothfield N. F., Schaller J. G., Talal N., Winchester R. J. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum. 1982 Nov;25(11):1271–1277. doi: 10.1002/art.1780251101. [DOI] [PubMed] [Google Scholar]
  40. Theofilopoulos A. N., Dixon F. J. Murine models of systemic lupus erythematosus. Adv Immunol. 1985;37:269–390. doi: 10.1016/s0065-2776(08)60342-9. [DOI] [PubMed] [Google Scholar]
  41. Tiegs S. L., Russell D. M., Nemazee D. Receptor editing in self-reactive bone marrow B cells. J Exp Med. 1993 Apr 1;177(4):1009–1020. doi: 10.1084/jem.177.4.1009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Yang P. M., Olsen N. J., Siminovitch K. A., Olee T., Kozin F., Carson D. A., Chen P. P. Possible deletion of a developmentally regulated heavy-chain variable region gene in autoimmune diseases. Proc Natl Acad Sci U S A. 1990 Oct;87(20):7907–7911. doi: 10.1073/pnas.87.20.7907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zimmer F. J., Hameister H., Schek H., Zachau H. G. Transposition of human immunoglobulin V kappa genes within the same chromosome and the mechanism of their amplification. EMBO J. 1990 May;9(5):1535–1542. doi: 10.1002/j.1460-2075.1990.tb08272.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zouali M., Madaio M. P., Canoso R. T., Stollar B. D. Restriction fragment length polymorphism analysis of the V kappa locus in human lupus. Eur J Immunol. 1989 Sep;19(9):1757–1760. doi: 10.1002/eji.1830190934. [DOI] [PubMed] [Google Scholar]

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