Skip to main content
NCBI home page
Annals of the Rheumatic Diseases logoLink to Annals of the Rheumatic Diseases
. 1995 Jun;54(6):471–476. doi: 10.1136/ard.54.6.471

Phospholipid binding specificities and idiotype expression of hybridoma derived monoclonal autoantibodies from splenic cells of patients with systemic lupus erythematosus.

C T Ravirajan 1, I Harmer 1, T McNally 1, A Hohmann 1, C G Mackworth-Young 1, D A Isenberg 1
PMCID: PMC1009905  PMID: 7632089

Abstract

OBJECTIVE--To analyse the phospholipid binding specificity, functional characteristics and idiotype expression of human hybridoma derived monoclonal autoantibodies (MAb) derived from the spleens of two patients with active systemic lupus erythematosus (SLE). METHODS--The IgM MAbs binding to phospholipids were generated from spleen cells of two patients (RSP and RT) with active SLE and their specificity of binding to neutral phospholipids (phosphatidyl ethanolamine, phosphatidyl choline, platelet activating factor, sphingomyelin) and negatively charged phospholipids (phosphatidyl glycerol, phosphatidyl serine, phosphatidic acid, phosphatidyl inositol and cardiolipin (CL)) analysed. Binding specificity of cross reactive antibodies (those binding to CL and DNA) was confirmed by fluid phase inhibition assays. Lupus anticoagulant activity and beta 2-glycoprotein-1 (beta 2 GP-1) requirement for the antigen binding of these MAbs were detected using the modified dilute Russell's viper venom test and modified anti-CL enzyme linked immunosorbent assay (ELISA), respectively. Expression of idiotypes (Id) Id RT-84 and Id H3 was analysed using rabbit polyclonal and murine monoclonal anti-idiotype reagents, respectively. RESULTS--Twelve clones from the patient RSP and eight clones from patient RT were reactive with phospholipids. Marked differences in phospholipid binding of these MAbs were noted, varying from truly polyreactive (RT-72 bound to most phospholipids tested) to monospecific (RT-84 bound only to CL). Furthermore, MAbs RT-84, RT-129, and RSP-57 had lupus anticoagulant activity and required beta 2 GP-1 for CL binding. It was found that 75% of phospholipid binding antibodies from RT clones expressed RT-84 Id, but none from RSP clones did so, and that Id H3 was expressed only by the RT-83 antibody. CONCLUSION--These results show that human anti-phospholipid MAbs are heterogeneous with respect to phospholipid binding, functional characteristics, and Id expression.

Full text

PDF
471

Selected References

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

  1. Brooks D. A., Bradley J., Zola H. A differentiation antigen expressed selectively by a proportion of human blood cells: detection with a monoclonal antibody. Pathology. 1982 Jan;14(1):5–11. doi: 10.3109/00313028209069036. [DOI] [PubMed] [Google Scholar]
  2. Carreras L. O., Defreyn G., Machin S. J., Vermylen J., Deman R., Spitz B., Van Assche A. Arterial thrombosis, intrauterine death and "lupus" antiocoagulant: detection of immunoglobulin interfering with prostacyclin formation. Lancet. 1981 Jan 31;1(8214):244–246. doi: 10.1016/s0140-6736(81)92087-0. [DOI] [PubMed] [Google Scholar]
  3. Casali P., Notkins A. L. Probing the human B-cell repertoire with EBV: polyreactive antibodies and CD5+ B lymphocytes. Annu Rev Immunol. 1989;7:513–535. doi: 10.1146/annurev.iy.07.040189.002501. [DOI] [PubMed] [Google Scholar]
  4. Costello P. B., Green F. A. Reactivity patterns of human anticardiolipin and other antiphospholipid antibodies in syphilitic sera. Infect Immun. 1986 Mar;51(3):771–775. doi: 10.1128/iai.51.3.771-775.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feinstein D. I. Lupus anticoagulant, thrombosis, and fetal loss. N Engl J Med. 1985 Nov 21;313(21):1348–1350. doi: 10.1056/NEJM198511213132109. [DOI] [PubMed] [Google Scholar]
  6. Galli M., Comfurius P., Maassen C., Hemker H. C., de Baets M. H., van Breda-Vriesman P. J., Barbui T., Zwaal R. F., Bevers E. M. Anticardiolipin antibodies (ACA) directed not to cardiolipin but to a plasma protein cofactor. Lancet. 1990 Jun 30;335(8705):1544–1547. doi: 10.1016/0140-6736(90)91374-j. [DOI] [PubMed] [Google Scholar]
  7. Harris E. N., Gharavi A. E., Boey M. L., Patel B. M., Mackworth-Young C. G., Loizou S., Hughes G. R. Anticardiolipin antibodies: detection by radioimmunoassay and association with thrombosis in systemic lupus erythematosus. Lancet. 1983 Nov 26;2(8361):1211–1214. doi: 10.1016/s0140-6736(83)91267-9. [DOI] [PubMed] [Google Scholar]
  8. Hohmann A., Comacchio R., Boswarva V., Sutjita M., Bradley J. The H3 anti-phospholipid idiotype is found in patients with systemic lupus erythematosus (SLE) but not in patients with syphilis. Clin Exp Immunol. 1991 Nov;86(2):207–211. doi: 10.1111/j.1365-2249.1991.tb05797.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hunt J. E., McNeil H. P., Morgan G. J., Crameri R. M., Krilis S. A. A phospholipid-beta 2-glycoprotein I complex is an antigen for anticardiolipin antibodies occurring in autoimmune disease but not with infection. Lupus. 1992 Feb;1(2):75–81. doi: 10.1177/096120339200100204. [DOI] [PubMed] [Google Scholar]
  10. Kalunian K. C., Peter J. B., Middlekauff H. R., Sayre J., Ando D. G., Mangotich M., Hahn B. H. Clinical significance of a single test for anti-cardiolipin antibodies in patients with systemic lupus erythematosus. Am J Med. 1988 Nov;85(5):602–608. doi: 10.1016/s0002-9343(88)80229-8. [DOI] [PubMed] [Google Scholar]
  11. Keeling D. M., Wilson A. J., Mackie I. J., Isenberg D. A., Machin S. J. Lupus anticoagulant activity of some antiphospholipid antibodies against phospholipid bound beta 2 glycoprotein I. J Clin Pathol. 1993 Jul;46(7):665–667. doi: 10.1136/jcp.46.7.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Koike T., Maruyama N., Funaki H., Tomioka H., Yoshida S. Specificity of mouse hybridoma antibodies to DNA. II. Phospholipid reactivity and biological false positive serological test for syphilis. Clin Exp Immunol. 1984 Aug;57(2):345–350. [PMC free article] [PubMed] [Google Scholar]
  13. Leung W. H., Wong K. L., Lau C. P., Wong C. K., Liu H. W. Association between antiphospholipid antibodies and cardiac abnormalities in patients with systemic lupus erythematosus. Am J Med. 1990 Oct;89(4):411–419. doi: 10.1007/BF01453668. [DOI] [PubMed] [Google Scholar]
  14. Loizou S., McCrea J. D., Rudge A. C., Reynolds R., Boyle C. C., Harris E. N. Measurement of anti-cardiolipin antibodies by an enzyme-linked immunosorbent assay (ELISA): standardization and quantitation of results. Clin Exp Immunol. 1985 Dec;62(3):738–745. [PMC free article] [PubMed] [Google Scholar]
  15. Love P. E., Santoro S. A. Antiphospholipid antibodies: anticardiolipin and the lupus anticoagulant in systemic lupus erythematosus (SLE) and in non-SLE disorders. Prevalence and clinical significance. Ann Intern Med. 1990 May 1;112(9):682–698. doi: 10.7326/0003-4819-112-9-682. [DOI] [PubMed] [Google Scholar]
  16. Matsuura E., Igarashi Y., Fujimoto M., Ichikawa K., Koike T. Anticardiolipin cofactor(s) and differential diagnosis of autoimmune disease. Lancet. 1990 Jul 21;336(8708):177–178. doi: 10.1016/0140-6736(90)91697-9. [DOI] [PubMed] [Google Scholar]
  17. Matsuura E., Igarashi Y., Fujimoto M., Ichikawa K., Suzuki T., Sumida T., Yasuda T., Koike T. Heterogeneity of anticardiolipin antibodies defined by the anticardiolipin cofactor. J Immunol. 1992 Jun 15;148(12):3885–3891. [PubMed] [Google Scholar]
  18. McNeil H. P., Simpson R. J., Chesterman C. N., Krilis S. A. Anti-phospholipid antibodies are directed against a complex antigen that includes a lipid-binding inhibitor of coagulation: beta 2-glycoprotein I (apolipoprotein H). Proc Natl Acad Sci U S A. 1990 Jun;87(11):4120–4124. doi: 10.1073/pnas.87.11.4120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Meroni P. L., Rivolta R., Ghidoni P. Histopathological findings in a case of systemic lupus erythematosus-associated anti-phospholipid syndrome. Clin Rheumatol. 1991 Jun;10(2):211–214. doi: 10.1007/BF02207667. [DOI] [PubMed] [Google Scholar]
  20. Out H. J., de Groot P. G., van Vliet M., de Gast G. C., Nieuwenhuis H. K., Derksen R. H. Antibodies to platelets in patients with anti-phospholipid antibodies. Blood. 1991 Jun 15;77(12):2655–2659. [PubMed] [Google Scholar]
  21. Radeke H. H., Christians U., Bleck J. S., Sewing K. F., Resch K. Additive and synergistic effects of cyclosporine metabolites on glomerular mesangial cells. Kidney Int. 1991 Jun;39(6):1255–1266. doi: 10.1038/ki.1991.159. [DOI] [PubMed] [Google Scholar]
  22. Rauch J., Janoff A. S. Phospholipid in the hexagonal II phase is immunogenic: evidence for immunorecognition of nonbilayer lipid phases in vivo. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4112–4114. doi: 10.1073/pnas.87.11.4112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rauch J., Tannenbaum M., Janoff A. S. Distinguishing plasma lupus anticoagulants from anti-factor antibodies using hexagonal (II) phase phospholipids. Thromb Haemost. 1989 Nov 24;62(3):892–896. [PubMed] [Google Scholar]
  24. Ravirajan C. T., Kalsi J., Wiloch H. W., Barakat S., Tuaillon N., Irvine W., Cockayne A., Harris A., Williams D. G., Williams W. Antigen-binding diversity of human hybridoma autoantibodies derived from splenocytes of patients with SLE. Lupus. 1992 May;1(3):157–165. doi: 10.1177/096120339200100307. [DOI] [PubMed] [Google Scholar]
  25. Sutjita M., Hohmann A., Comacchio R., Boey M. L., Bradley J. A common anti-cardiolipin antibody idiotype in autoimmune disease: identification using a mouse monoclonal antibody directed against a naturally-occurring anti-phospholipid antibody. Clin Exp Immunol. 1989 Feb;75(2):211–216. [PMC free article] [PubMed] [Google Scholar]
  26. Sutjita M., Hohmann A., Comacchio R., Bradley J. Polyspecific human and murine antibodies to diphtheria and tetanus toxoids and phospholipids. Clin Exp Immunol. 1988 Aug;73(2):191–197. [PMC free article] [PubMed] [Google Scholar]
  27. Watts R. A., Williams W., Le Page S., Norden A., Soltys A., Swana G., Addison I., Hay F. C., Isenberg D. A. Analysis of autoantibody reactivity and common idiotype PR4 expression of myeloma proteins. J Autoimmun. 1989 Oct;2(5):689–700. doi: 10.1016/s0896-8411(89)80007-1. [DOI] [PubMed] [Google Scholar]

Articles from Annals of the Rheumatic Diseases are provided here courtesy of BMJ Publishing Group

RESOURCES