Skip to main content
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1987 Jun;68(3):552–561.

The effect of La(SSB) on PWM-induced immunoglobulin synthesis by anti-La(SSB) positive SLE patients and healthy controls.

G Smith 1, C A Morrison 1, P J Maddison 1
PMCID: PMC1542759  PMID: 2958189

Abstract

The effect of affinity purified La(SSB) on immunoglobulin synthesis in vitro by mononuclear cells (MNC) from anti-La(SSB)-positive systemic lupus erythematosus (SLE) patients and healthy controls was studied. La(SSB) was prepared from calf thymus extract and characterized by SDS-polyacrylamide gel electrophoresis and immunoblotting. Silver staining of gels reveals nine major bands at 68 kD, 43-48 kD and 30-33 kD, of which six were recognized on immunoblots by sera from anti-La(SSB) positive SLE patients. Studies in vitro showed that La(SSB) alone did not stimulate total IgG or IgM synthesis in controls or SLE patients. Low concentrations of La(SSB) (optimal dose less than 0.02 ng/ml) suppressed pokeweed mitogen (PWM)-driven IgG synthesis by controls but not by anti-La(SSB) positive SLE patients. IgM responses were unaffected. Anti-La(SSB) and anti-DNA were detected in PWM-stimulated cultures from both study groups. In the presence of La(SSB) IgM anti-La(SSB) synthesis was enhanced in anti-La(SSB)-positive patients in a dose-dependent manner. In contrast, La(SSB) inhibited anti-La(SSB) production by controls (maximal at 2 ng/ml). La(SSB) had no effect on anti-DNA production in either group. Pre-incubation of control or anti-La(SSB)-positive SLE MNC with La(SSB) before addition to autologous PWM-driven cultures did not induce suppressor cells, although pre-incubation with Concanavalin A (ConA) did. Thus we suggest that La(SSB)-induced suppression of IgG synthesis in PWM-driven control cultures may not be due to induction of regulator cells, possibly missing from SLE cultures, but perhaps is a direct effect on B cells.

Full text

PDF
552

Images in this article

Selected References

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

  1. Alarcón-Segovia D. Cellular immunity and its regulation in SLE. Clin Rheum Dis. 1982 Apr;8(1):63–75. [PubMed] [Google Scholar]
  2. Birdsall H. H., Rossen R. D. Regulation of antibody release by naturally occurring anti-immunoglobulins in cultures of pokeweed mitogen-stimulated human peripheral blood lymphocytes. J Immunol. 1984 Sep;133(3):1257–1264. [PubMed] [Google Scholar]
  3. Clough J. D., Frank S. A., Calabrese L. H. Deficiency of T cell mediated regulation of anti-DNA production in systemic lupus erythematosus. Arthritis Rheum. 1980 Jan;23(1):24–29. doi: 10.1002/art.1780230105. [DOI] [PubMed] [Google Scholar]
  4. Duran N., Bach M., Puigdomenech P., Palau J. Characterization of antigenic polypeptides of the RNP, Sm and SS-B nuclear antigens from calf thymus. Mol Immunol. 1984 Aug;21(8):731–739. doi: 10.1016/0161-5890(84)90026-9. [DOI] [PubMed] [Google Scholar]
  5. Eichmann K. Idiotype suppression. I. Influence of the dose and of the effector functions of anti-idiotypic antibody on the production of an idiotype. Eur J Immunol. 1974 Apr;4(4):296–302. doi: 10.1002/eji.1830040413. [DOI] [PubMed] [Google Scholar]
  6. Eschenbruch M., Bürk R. R. Experimentally improved reliability of ultrasensitive silver staining of protein in polyacrylamide gels. Anal Biochem. 1982 Sep 1;125(1):96–99. doi: 10.1016/0003-2697(82)90387-6. [DOI] [PubMed] [Google Scholar]
  7. Francoeur A. M., Mathews M. B. Interaction between VA RNA and the lupus antigen La: formation of a ribonucleoprotein particle in vitro. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6772–6776. doi: 10.1073/pnas.79.22.6772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Geha R. S. Presence of auto-anti-idiotypic antibody during the normal human immune response to tetanus toxoid antigen. J Immunol. 1982 Jul;129(1):139–144. [PubMed] [Google Scholar]
  9. Habets W. J., den Brok J. H., Boerbooms A. M., van de Putte L. B., van Venrooij W. J. Characterization of the SS-B (La) antigen in adenovirus-infected and uninfected HeLa cells. EMBO J. 1983;2(10):1625–1631. doi: 10.1002/j.1460-2075.1983.tb01636.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hardin J. A., Rahn D. R., Shen C., Lerner M. R., Wolin S. L., Rosa M. D., Steitz J. A. Antibodies from patients with connective tissue diseases bind specific subsets of cellular RNA-protein particles. J Clin Invest. 1982 Jul;70(1):141–147. doi: 10.1172/JCI110587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoch S., Schur P. H., Schwaber J. Frequency of anti-DNA antibody producing cells from normals and patients with systemic lupus erythematosus. Clin Immunol Immunopathol. 1983 Apr;27(1):28–37. doi: 10.1016/0090-1229(83)90053-3. [DOI] [PubMed] [Google Scholar]
  12. Kallenberg C. G., Limburg P. C., Van Slochteren C., Van der Woude F. J., The T. H. B cell activity in systemic lupus erythematosus: depressed in vivo humoral immune response to a primary antigen (haemocyanin) and increased in vitro spontaneous immunoglobulin synthesis. Clin Exp Immunol. 1983 Aug;53(2):371–383. [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Mach P. S., Kharouby M., Lutcher F., Olivier J. L., Bazely N., Dougados M., Amor B. The in vitro production and regulation of anti-double stranded DNA antibodies by peripheral blood mononuclear cells from normals and patients with systemic lupus erythematosus. Clin Exp Immunol. 1984 Aug;57(2):338–344. [PMC free article] [PubMed] [Google Scholar]
  15. Maddison P. J., Skinner R. P., Vlachoyiannopoulos P., Brennand D. M., Hough D. Antibodies to nRNP, Sm, Ro(SSA) and La(SSB) detected by ELISA: their specificity and inter-relations in connective tissue disease sera. Clin Exp Immunol. 1985 Nov;62(2):337–345. [PMC free article] [PubMed] [Google Scholar]
  16. Morimoto C., Abe T., Homma M. Altered function of suppressor T lymphocytes in patients with active systemic lupus erythematosus--in vitro immune response to autoantigen. Clin Immunol Immunopathol. 1979 Jun;13(2):161–170. doi: 10.1016/0090-1229(79)90060-6. [DOI] [PubMed] [Google Scholar]
  17. Mumford P., Room G. R., Venables P. J., Maini R. N. IgG antibodies to SS-B (La), RNP/Sm and DNA are produced by PWM-stimulated normal human lymphocytes in culture. Rheumatol Int. 1985;5(3):109–112. doi: 10.1007/BF00541329. [DOI] [PubMed] [Google Scholar]
  18. Rauch J., Tannenbaum H., Stollar B. D., Schwartz R. S. Monoclonal anti-cardiolipin antibodies bind to DNA. Eur J Immunol. 1984 Jun;14(6):529–534. doi: 10.1002/eji.1830140609. [DOI] [PubMed] [Google Scholar]
  19. Rinke J., Steitz J. A. Precursor molecules of both human 5S ribosomal RNA and transfer RNAs are bound by a cellular protein reactive with anti-La lupus antibodies. Cell. 1982 May;29(1):149–159. doi: 10.1016/0092-8674(82)90099-x. [DOI] [PubMed] [Google Scholar]
  20. Saxon A., Barnett E. Human auto-antiidiotypes regulating T cell-mediated reactivity to tetanus toxoid. J Clin Invest. 1984 Feb;73(2):342–348. doi: 10.1172/JCI111218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smith G., Skinner R. P., Bliss J., Maddison P. J. In vitro autoantibody synthesis by anti-La(SSB) positive SLE patients and healthy controls. Clin Exp Rheumatol. 1986 Jul-Sep;4(3):209–216. [PubMed] [Google Scholar]
  22. Stevens R. H. Immunoglobulin-bearing cells are a target for the antigen-induced inhibition of pokeweed mitogen-stimulated antibody production. J Immunol. 1981 Sep;127(3):968–972. [PubMed] [Google Scholar]
  23. 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]
  24. Tardieu M., Powers M. L., Hafler D. A., Hauser S. L., Weiner H. L. Autoimmunity following viral infection: demonstration of monoclonal antibodies against normal tissue following infection of mice with reovirus and demonstration of shared antigenicity between virus and lymphocytes. Eur J Immunol. 1984 Jun;14(6):561–565. doi: 10.1002/eji.1830140614. [DOI] [PubMed] [Google Scholar]
  25. Venables P. J., Charles P. J., Buchanan R. R., Yi T., Mumford P. A., Schrieber L., Room G. R., Maini R. N. Quantitation and detection of isotypes of anti-SS-B antibodies by ELISA and Farr assays using affinity purified antigens. An approach to the investigation of Sjögren's syndrome and systemic lupus erythematosus. Arthritis Rheum. 1983 Feb;26(2):146–155. doi: 10.1002/art.1780260205. [DOI] [PubMed] [Google Scholar]
  26. Volkman D. J., Allyn S. P., Fauci A. S. Antigen-induced in vitro antibody production in humans: tetanus toxoid-specific antibody synthesis. J Immunol. 1982 Jul;129(1):107–112. [PubMed] [Google Scholar]
  27. Volkman D. J., Lane H. C., Fauci A. S. Antigen-induced in vitro antibody production in humans: a model for B cell activation and immunoregulation. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2528–2531. doi: 10.1073/pnas.78.4.2528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zouali M., Eyquem A. Expression of anti-idiotypic clones against auto-anti-DNA antibodies in normal individuals. Cell Immunol. 1983 Feb 15;76(1):137–147. doi: 10.1016/0008-8749(83)90356-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES