Skip to main content
PMC home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1993 Nov;94(2):385–390. doi: 10.1111/j.1365-2249.1993.tb03461.x

Characterization of a human monoclonal autoantibody directed to cardiolipin/beta 2 glycoprotein I produced by chronic lymphocytic leukaemia B cells.

X Mariette 1, Y Levy 1, M L Dubreuil 1, L Intrator 1, F Danon 1, J C Brouet 1
PMCID: PMC1534242  PMID: 8222331

Abstract

We determined the specificity and sequence of immunoglobulin molecules synthesized by monoclonal B cells from a patient with chronic lymphocytic leukaemia (CLL) who presented with a number of clinical and biological autoimmune symptoms. Heterohybrids obtained by fusion of CLL cells with the mouse X63-Ag 8.653 myeloma produced IgM lambda MoAbs directed to the cardiolipin/beta 2 glycoprotein I (beta 2GPI) complex and ssDNA. They were devoid of polyreactivity. Nucleotide sequence analysis of the variable domain of the mu chain indicated the utilization of the VH4 71.2 gene or one allotypic variant, DXP4 and JH3 segments. The lambda light chain used the single gene from the V lambda 8 subfamily, J lambda 3 and C lambda 3 genes. The VH gene displayed 11 nucleotide changes in comparison with its putative germline counterpart. However, these nucleotide changes correspond to variations observed in other published VH4 sequences, suggesting gene polymorphism rather than somatic mutation. DXP4 and JH3 were also in germline configuration. The VL gene exhibited a single replacement mutation in CDR1. These data suggest that the monoclonal CLL B cells in this patient retained VH and VL genes in germline configuration although they secreted a pathogenic anti-cardiolipin antibody associated with clinical symptoms, vasculitis and thrombosis, which may be provoked by antibodies to the phospholipid/beta 2GPI complex.

Full text

PDF
390

Images in this article

387Fig. 1
on p.387

Selected References

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

  1. Bertoli L. F., Kubagawa H., Borzillo G. V., Burrows P. D., Schreeder M. T., Carroll A. J., Cooper M. D. Bone marrow origin of a B-cell lymphoma. Blood. 1988 Jul;72(1):94–101. [PubMed] [Google Scholar]
  2. Borche L., Lim A., Binet J. L., Dighiero G. Evidence that chronic lymphocytic leukemia B lymphocytes are frequently committed to production of natural autoantibodies. Blood. 1990 Aug 1;76(3):562–569. [PubMed] [Google Scholar]
  3. Cai J., Humphries C., Richardson A., Tucker P. W. Extensive and selective mutation of a rearranged VH5 gene in human B cell chronic lymphocytic leukemia. J Exp Med. 1992 Oct 1;176(4):1073–1081. doi: 10.1084/jem.176.4.1073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Casali P., Burastero S. E., Nakamura M., Inghirami G., Notkins A. L. Human lymphocytes making rheumatoid factor and antibody to ssDNA belong to Leu-1+ B-cell subset. Science. 1987 Apr 3;236(4797):77–81. doi: 10.1126/science.3105056. [DOI] [PubMed] [Google Scholar]
  5. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  6. Deane M., Baker B. W., Norton J. D. Immunoglobulin VH4 gene usage in B lymphoid leukaemias. Br J Haematol. 1993 Jun;84(2):242–249. doi: 10.1111/j.1365-2141.1993.tb03059.x. [DOI] [PubMed] [Google Scholar]
  7. Dersimonian H., Schwartz R. S., Barrett K. J., Stollar B. D. Relationship of human variable region heavy chain germ-line genes to genes encoding anti-DNA autoantibodies. J Immunol. 1987 Oct 1;139(7):2496–2501. [PubMed] [Google Scholar]
  8. 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]
  9. Hough D. W., Eady R. P., Hamblin T. J., Stevenson F. K., Stevenson G. T. Anti-idiotype sera raised against surface immunoglobulin of human neoplastic lymphocytes. J Exp Med. 1976 Oct 1;144(4):960–969. doi: 10.1084/jem.144.4.960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Humphries C. G., Shen A., Kuziel W. A., Capra J. D., Blattner F. R., Tucker P. W. A new human immunoglobulin VH family preferentially rearranged in immature B-cell tumours. Nature. 1988 Feb 4;331(6155):446–449. doi: 10.1038/331446a0. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Kipps T. J., Tomhave E., Chen P. P., Carson D. A. Autoantibody-associated kappa light chain variable region gene expressed in chronic lymphocytic leukemia with little or no somatic mutation. Implications for etiology and immunotherapy. J Exp Med. 1988 Mar 1;167(3):840–852. doi: 10.1084/jem.167.3.840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Koike T., Sueishi M., Funaki H., Tomioka H., Yoshida S. Anti-phospholipid antibodies and biological false positive serological test for syphilis in patients with systemic lupus erythematosus. Clin Exp Immunol. 1984 Apr;56(1):193–199. [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Logtenberg T., Schutte M. E., Ebeling S. B., Gmelig-Meyling F. H., van Es J. H. Molecular approaches to the study of human B-cell and (auto)antibody repertoire generation and selection. Immunol Rev. 1992 Aug;128:23–47. doi: 10.1111/j.1600-065x.1992.tb00831.x. [DOI] [PubMed] [Google Scholar]
  16. Logtenberg T., Schutte M. E., Inghirami G., Berman J. E., Gmelig-Meyling F. H., Insel R. A., Knowles D. M., Alt F. W. Immunoglobulin VH gene expression in human B cell lines and tumors: biased VH gene expression in chronic lymphocytic leukemia. Int Immunol. 1989;1(4):362–366. doi: 10.1093/intimm/1.4.362. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Martin T., Duffy S. F., Carson D. A., Kipps T. J. Evidence for somatic selection of natural autoantibodies. J Exp Med. 1992 Apr 1;175(4):983–991. doi: 10.1084/jem.175.4.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Meeker T. C., Grimaldi J. C., O'Rourke R., Loeb J., Juliusson G., Einhorn S. Lack of detectable somatic hypermutation in the V region of the Ig H chain gene of a human chronic B lymphocytic leukemia. J Immunol. 1988 Dec 1;141(11):3994–3998. [PubMed] [Google Scholar]
  20. Olee T., Lu E. W., Huang D. F., Soto-Gil R. W., Deftos M., Kozin F., Carson D. A., Chen P. P. Genetic analysis of self-associating immunoglobulin G rheumatoid factors from two rheumatoid synovia implicates an antigen-driven response. J Exp Med. 1992 Mar 1;175(3):831–842. doi: 10.1084/jem.175.3.831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pascual V., Capra J. D. VH4-21, a human VH gene segment overrepresented in the autoimmune repertoire. Arthritis Rheum. 1992 Jan;35(1):11–18. doi: 10.1002/art.1780350103. [DOI] [PubMed] [Google Scholar]
  22. Preud'homme J. L., Rochard E., Gouet D., Danon F., Alcalay M., Touchard G., Aucouturier P. Isotypic distribution of anti-double-stranded DNA antibodies: a diagnostic evaluation by enzyme-linked immunosorbent assay. Diagn Clin Immunol. 1988;5(5):256–261. [PubMed] [Google Scholar]
  23. Preud'homme J. L., Seligmann M. Surface bound immunoglobulins as a cell marker in human lymphoproliferative diseases. Blood. 1972 Dec;40(6):777–794. [PubMed] [Google Scholar]
  24. Rassenti L. Z., Kipps T. J. Lack of extensive mutations in the VH5 genes used in common B cell chronic lymphocytic leukemia. J Exp Med. 1993 Apr 1;177(4):1039–1046. doi: 10.1084/jem.177.4.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Roubey R. A., Pratt C. W., Buyon J. P., Winfield J. B. Lupus anticoagulant activity of autoimmune antiphospholipid antibodies is dependent upon beta 2-glycoprotein I. J Clin Invest. 1992 Sep;90(3):1100–1104. doi: 10.1172/JCI115926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanz I., Casali P., Thomas J. W., Notkins A. L., Capra J. D. Nucleotide sequences of eight human natural autoantibody VH regions reveals apparent restricted use of VH families. J Immunol. 1989 Jun 1;142(11):4054–4061. [PubMed] [Google Scholar]
  28. 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]
  29. Siminovitch K. A., Misener V., Kwong P. C., Song Q. L., Chen P. P. A natural autoantibody is encoded by germline heavy and lambda light chain variable region genes without somatic mutation. J Clin Invest. 1989 Nov;84(5):1675–1678. doi: 10.1172/JCI114347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Siminovitch K. A., Misener V., Kwong P. C., Yang P. M., Laskin C. A., Cairns E., Bell D., Rubin L. A., Chen P. P. A human anti-cardiolipin autoantibody is encoded by developementally restricted heavy and light chain variable region genes. Autoimmunity. 1990;8(2):97–105. doi: 10.3109/08916939008995727. [DOI] [PubMed] [Google Scholar]
  31. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Tomlinson I. M., Walter G., Marks J. D., Llewelyn M. B., Winter G. The repertoire of human germline VH sequences reveals about fifty groups of VH segments with different hypervariable loops. J Mol Biol. 1992 Oct 5;227(3):776–798. doi: 10.1016/0022-2836(92)90223-7. [DOI] [PubMed] [Google Scholar]
  34. Tsapis A., Bentaboulet M., Pellet P., Mihaesco E., Thierry D., Seligmann M., Brouet J. C. The productive gene for alpha-H chain disease protein MAL is highly modified by insertion-deletion processes. J Immunol. 1989 Dec 1;143(11):3821–3827. [PubMed] [Google Scholar]
  35. Van Es J. H., Aanstoot H., Gmelig-Meyling F. H., Derksen R. H., Logtenberg T. A human systemic lupus erythematosus-related anti-cardiolipin/single-stranded DNA autoantibody is encoded by a somatically mutated variant of the developmentally restricted 51P1 VH gene. J Immunol. 1992 Sep 15;149(6):2234–2240. [PubMed] [Google Scholar]
  36. Viard J. P., Amoura Z., Bach J. F. Association of anti-beta 2 glycoprotein I antibodies with lupus-type circulating anticoagulant and thrombosis in systemic lupus erythematosus. Am J Med. 1992 Aug;93(2):181–186. doi: 10.1016/0002-9343(92)90049-h. [DOI] [PubMed] [Google Scholar]
  37. Weng N. P., Snyder J. G., Yu-Lee L. Y., Marcus D. M. Polymorphism of human immunoglobulin VH4 germ-line genes. Eur J Immunol. 1992 Apr;22(4):1075–1082. doi: 10.1002/eji.1830220430. [DOI] [PubMed] [Google Scholar]
  38. Winkler T. H., Fehr H., Kalden J. R. Analysis of immunoglobulin variable region genes from human IgG anti-DNA hybridomas. Eur J Immunol. 1992 Jul;22(7):1719–1728. doi: 10.1002/eji.1830220709. [DOI] [PubMed] [Google Scholar]
  39. van Es J. H., Heutink M., Aanstoot H., Logtenberg T. Sequence analysis of members of the human Ig VH4 gene family derived from a single VH locus. Identification of novel germ-line members. J Immunol. 1992 Jul 15;149(2):492–497. [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES