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. 1994 Feb;93(2):718–724. doi: 10.1172/JCI117025

Cellular mechanism of the conduction abnormalities induced by serum from anti-Ro/SSA-positive patients in rabbit hearts.

S Garcia 1, J H Nascimento 1, E Bonfa 1, R Levy 1, S F Oliveira 1, A V Tavares 1, A C de Carvalho 1
PMCID: PMC293909  PMID: 8113406

Abstract

In this study, IgG fractions from sera of SLE patients with anti-Ro/SSA or anti-Ro/SSA and anti-La/SSB activity were tested in Langendorff preparations of adult rabbit hearts, aiming to reproduce the cardiac manifestations observed in neonatal lupus in an experimental model. The hearts were perfused with normal Tyrode's solution for 30 min, followed by perfusion with Tyrode's containing 0.3 mg/ml of anti-Ro/SSA- (or anti-Ro/La-) positive IgG (nine sera), anti-ribonucleoprotein (RNP)-positive IgG (five sera), or IgG fractions from normal donors (five sera). In one third of the experiments done with anti-Ro/La-positive IgG, heart block was observed. With the remaining fractions, a decrease in heart rate of 17.1% was observed, but normal sinus rhythm was maintained. The IgG fractions with anti-RNP activity (five experiments) and from normal sera (six experiments) reduced heart rates by 12.9 and 3.3%, respectively, but heart block was not observed. To further characterize the cellular mechanisms involved in the conduction disturbances observed in the whole rabbit hearts, we conducted experiments with ventricular myocytes isolated from young rabbit hearts, studied by whole cell patch-clamp technique. In these experiments, the slow inward currents were analyzed during the superfusion of the cell with normal Tyrode's solution and 5 min after superfusion with Tyrode's solution containing 0.3 mg/ml of anti-Ro/SSA- (or anti-Ro/La-) positive IgG (five sera), anti-RNP-positive IgG (three sera), or IgG from normal donors (four sera). Resting and action potential amplitudes were not affected by any of the sera used. The anti-Ro/SSA IgG fraction induced a mean reduction in the peak slow inward current of 31.6%. IgG fractions with anti-RNP activity reduced slow inward current by 4.4%, whereas IgG fractions from normal donors increased this current by 3.3%. IgG-free fractions from sera of patients with anti-Ro/SSA activity did not alter the peak slow inward current. These results show, for the first time, that the presence of anti-Ro/SSA or anti-Ro/SSA and anti-La/SSB antibody activity in IgG fractions from lupus patients' sera can induce cardiac conduction disorders similar to those observed in neonatal lupus.

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Selected References

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

  1. Alexander E., Buyon J. P., Provost T. T., Guarnieri T. Anti-Ro/SS-A antibodies in the pathophysiology of congenital heart block in neonatal lupus syndrome, an experimental model. In vitro electrophysiologic and immunocytochemical studies. Arthritis Rheum. 1992 Feb;35(2):176–189. doi: 10.1002/art.1780350209. [DOI] [PubMed] [Google Scholar]
  2. Bachmann M., Pfeifer K., Schröder H. C., Müller W. E. Characterization of the autoantigen La as a nucleic acid-dependent ATPase/dATPase with melting properties. Cell. 1990 Jan 12;60(1):85–93. doi: 10.1016/0092-8674(90)90718-t. [DOI] [PubMed] [Google Scholar]
  3. Behan W. M., Behan P. O., Reid J. M., Doig W., Gairns J. Family studies of congenital heart block associated with Ro antibody. Br Heart J. 1989 Oct;62(4):320–324. doi: 10.1136/hrt.62.4.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ben-Chetrit E., Gandy B. J., Tan E. M., Sullivan K. F. Isolation and characterization of a cDNA clone encoding the 60-kD component of the human SS-A/Ro ribonucleoprotein autoantigen. J Clin Invest. 1989 Apr;83(4):1284–1292. doi: 10.1172/JCI114013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bilazarian S. D., Taylor A. J., Brezinski D., Hochberg M. C., Guarnieri T., Provost T. T. High-grade atrioventricular heart block in an adult with systemic lupus erythematosus: the association of nuclear RNP (U1 RNP) antibodies, a case report, and review of the literature. Arthritis Rheum. 1989 Sep;32(9):1170–1174. doi: 10.1002/anr.1780320918. [DOI] [PubMed] [Google Scholar]
  6. Boire G., Lopez-Longo F. J., Lapointe S., Ménard H. A. Sera from patients with autoimmune disease recognize conformational determinants on the 60-kd Ro/SS-A protein. Arthritis Rheum. 1991 Jun;34(6):722–730. doi: 10.1002/art.1780340613. [DOI] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Buyon J. P., Winchester R. Congenital complete heart block. A human model of passively acquired autoimmune injury. Arthritis Rheum. 1990 May;33(5):609–614. doi: 10.1002/art.1780330502. [DOI] [PubMed] [Google Scholar]
  9. Chambers J. C., Kenan D., Martin B. J., Keene J. D. Genomic structure and amino acid sequence domains of the human La autoantigen. J Biol Chem. 1988 Dec 5;263(34):18043–18051. [PubMed] [Google Scholar]
  10. Chan E. K., Hamel J. C., Buyon J. P., Tan E. M. Molecular definition and sequence motifs of the 52-kD component of human SS-A/Ro autoantigen. J Clin Invest. 1991 Jan;87(1):68–76. doi: 10.1172/JCI115003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cohen N. M., Lederer W. J. Changes in the calcium current of rat heart ventricular myocytes during development. J Physiol. 1988 Dec;406:115–146. doi: 10.1113/jphysiol.1988.sp017372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Deutscher S. L., Harley J. B., Keene J. D. Molecular analysis of the 60-kDa human Ro ribonucleoprotein. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9479–9483. doi: 10.1073/pnas.85.24.9479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Elkon K. B., Jankowski P. W. Fine specificities of autoantibodies directed against the Ro, La, Sm, RNP, and Jo-1 proteins defined by two-dimensional gel electrophoresis and immunoblotting. J Immunol. 1985 Jun;134(6):3819–3824. [PubMed] [Google Scholar]
  14. Esscher E., Scott J. S. Congenital heart block and maternal systemic lupus erythematosus. Br Med J. 1979 May 12;1(6173):1235–1238. doi: 10.1136/bmj.1.6173.1235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Furukawa F., Kashihara-Sawami M., Lyons M. B., Norris D. A. Binding of antibodies to the extractable nuclear antigens SS-A/Ro and SS-B/La is induced on the surface of human keratinocytes by ultraviolet light (UVL): implications for the pathogenesis of photosensitive cutaneous lupus. J Invest Dermatol. 1990 Jan;94(1):77–85. doi: 10.1111/1523-1747.ep12873930. [DOI] [PubMed] [Google Scholar]
  16. Furukawa F., Lyons M. B., Lee L. A., Coulter S. N., Norris D. A. Estradiol enhances binding to cultured human keratinocytes of antibodies specific for SS-A/Ro and SS-B/La. Another possible mechanism for estradiol influence of lupus erythematosus. J Immunol. 1988 Sep 1;141(5):1480–1488. [PubMed] [Google Scholar]
  17. Geggel R. L., Tucker L., Szer I. Postnatal progression from second- to third-degree heart block in neonatal lupus syndrome. J Pediatr. 1988 Dec;113(6):1049–1052. doi: 10.1016/s0022-3476(88)80581-x. [DOI] [PubMed] [Google Scholar]
  18. Giles W. R., Imaizumi Y. Comparison of potassium currents in rabbit atrial and ventricular cells. J Physiol. 1988 Nov;405:123–145. doi: 10.1113/jphysiol.1988.sp017325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  20. Harley J. B., Kaine J. L., Fox O. F., Reichlin M., Gruber B. Ro (SS-A) antibody and antigen in a patient with congenital complete heart block. Arthritis Rheum. 1985 Dec;28(12):1321–1325. doi: 10.1002/art.1780281202. [DOI] [PubMed] [Google Scholar]
  21. Kim Y. I., Neher E. IgG from patients with Lambert-Eaton syndrome blocks voltage-dependent calcium channels. Science. 1988 Jan 22;239(4838):405–408. doi: 10.1126/science.2447652. [DOI] [PubMed] [Google Scholar]
  22. Kurata N., Tan E. M. Identification of antibodies to nuclear acidic antigens by counterimmunoelectrophoresis. Arthritis Rheum. 1976 May-Jun;19(3):574–580. doi: 10.1002/art.1780190309. [DOI] [PubMed] [Google Scholar]
  23. Lee L. A., Gaither K. K., Coulter S. N., Norris D. A., Harley J. B. Pattern of cutaneous immunoglobulin G deposition in subacute cutaneous lupus erythematosus is reproduced by infusing purified anti-Ro (SSA) autoantibodies into human skin-grafted mice. J Clin Invest. 1989 May;83(5):1556–1562. doi: 10.1172/JCI114052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lee L. A., Weston W. L., Krueger G. G., Emam M., Reichlin M., Stevens J. O., Surbrugg S. K., Vasil A., Norris D. A. An animal model of antibody binding in cutaneous lupus. Arthritis Rheum. 1986 Jun;29(6):782–788. doi: 10.1002/art.1780290612. [DOI] [PubMed] [Google Scholar]
  25. Lieu T. S., Newkirk M. M., Arnett F. C., Lee L. A., Deng J. S., Capra J. D., Sontheimer R. D. A major autoepitope is present on the amino terminus of a human SS-A/Ro polypeptide. J Autoimmun. 1989 Aug;2(4):367–374. doi: 10.1016/0896-8411(89)90165-0. [DOI] [PubMed] [Google Scholar]
  26. Lockshin M. D., Druzin M. L., Goei S., Qamar T., Magid M. S., Jovanovic L., Ferenc M. Antibody to cardiolipin as a predictor of fetal distress or death in pregnant patients with systemic lupus erythematosus. N Engl J Med. 1985 Jul 18;313(3):152–156. doi: 10.1056/NEJM198507183130304. [DOI] [PubMed] [Google Scholar]
  27. Magsaam J., Gharavi A. E., Parnassa A. P., Weissbach H., Brot N., Elkon K. B. Quantification of lupus anti-ribosome P antibodies using a recombinant P2 fusion protein and determination of the predicted amino acid sequence of the autoantigen in patients' mononuclear cells. Clin Exp Immunol. 1989 May;76(2):165–171. [PMC free article] [PubMed] [Google Scholar]
  28. Masuda M. O., de Magalhães Engel G., Barbosa Moreira A. P. Characterization of isolated ventricular myocytes: two levels of resting potential. J Mol Cell Cardiol. 1987 Sep;19(9):831–839. doi: 10.1016/s0022-2828(87)80612-0. [DOI] [PubMed] [Google Scholar]
  29. Mayet W. J., Bachmann M., Pfeifer K., Schröder H. C., Müller W. E., Gudat W., Korting G. W., Meyer zum Büschenfelde K. H. A monoclonal Ro-antibody and the serum of a Ro-positive patient with subacute cutaneous lupus erythematosus (SCLE) react with basal layers of human epidermis. Eur J Clin Invest. 1988 Oct;18(5):465–471. doi: 10.1111/j.1365-2362.1988.tb01041.x. [DOI] [PubMed] [Google Scholar]
  30. McCauliffe D. P., Lux F. A., Lieu T. S., Sanz I., Hanke J., Newkirk M. M., Bachinski L. L., Itoh Y., Siciliano M. J., Reichlin M. Molecular cloning, expression, and chromosome 19 localization of a human Ro/SS-A autoantigen. J Clin Invest. 1990 May;85(5):1379–1391. doi: 10.1172/JCI114582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. McCauliffe D. P., Zappi E., Lieu T. S., Michalak M., Sontheimer R. D., Capra J. D. A human Ro/SS-A autoantigen is the homologue of calreticulin and is highly homologous with onchocercal RAL-1 antigen and an aplysia "memory molecule". J Clin Invest. 1990 Jul;86(1):332–335. doi: 10.1172/JCI114704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. McCue C. M., Mantakas M. E., Tingelstad J. B., Ruddy S. Congenital heart block in newborns of mothers with connective tissue disease. Circulation. 1977 Jul;56(1):82–90. doi: 10.1161/01.cir.56.1.82. [DOI] [PubMed] [Google Scholar]
  33. McCune A. B., Weston W. L., Lee L. A. Maternal and fetal outcome in neonatal lupus erythematosus. Ann Intern Med. 1987 Apr;106(4):518–523. doi: 10.7326/0003-4819-106-4-518. [DOI] [PubMed] [Google Scholar]
  34. Meilof J. F., Bantjes I., De Jong J., Van Dam A. P., Smeenk R. J. The detection of anti-Ro/SS-A and anti-La/SS-B antibodies. A comparison of counterimmunoelectrophoresis with immunoblot, ELISA, and RNA-precipitation assays. J Immunol Methods. 1990 Oct 19;133(2):215–226. doi: 10.1016/0022-1759(90)90362-y. [DOI] [PubMed] [Google Scholar]
  35. Miyagawa S., Dohi K., Yoshioka A., Shirai T. Female predominance of immune response to SSA/Ro antigens and risk of neonatal lupus erythematosus. Br J Dermatol. 1990 Aug;123(2):223–227. doi: 10.1111/j.1365-2133.1990.tb01850.x. [DOI] [PubMed] [Google Scholar]
  36. Rokeach L. A., Haselby J. A., Meilof J. F., Smeenk R. J., Unnasch T. R., Greene B. M., Hoch S. O. Characterization of the autoantigen calreticulin. J Immunol. 1991 Nov 1;147(9):3031–3039. [PubMed] [Google Scholar]
  37. Sontheimer R. D., McCauliffe D. P. Pathogenesis of anti-Ro/SS-A autoantibody-associated cutaneous lupus erythematosus. Dermatol Clin. 1990 Oct;8(4):751–758. [PubMed] [Google Scholar]
  38. Tan E. M. Autoantibodies to nuclear antigens (ANA): their immunobiology and medicine. Adv Immunol. 1982;33:167–240. doi: 10.1016/s0065-2776(08)60836-6. [DOI] [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. Watson R. M., Braunstein B. L., Watson A. J., Hochberg M. C., Provost T. T. Fetal wastage in women with anti-Ro(SSA) antibody. J Rheumatol. 1986 Feb;13(1):90–94. [PubMed] [Google Scholar]

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