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. 1994 Apr 1;179(4):1317–1330. doi: 10.1084/jem.179.4.1317

Autoantigens targeted in systemic lupus erythematosus are clustered in two populations of surface structures on apoptotic keratinocytes

PMCID: PMC2191465  PMID: 7511686

Abstract

Systemic lupus erythematosus is a multisystem autoimmune disease in which the autoantibody response targets a variety of autoantigens of diverse subcellular location. We show here that these autoantigens are clustered in two distinct populations of blebs at the surface of apoptotic cells. The population of smaller blebs contains fragmented endoplasmic reticulum (ER) and ribosomes, as well as the ribonucleoprotein, Ro. The larger blebs (apoptotic bodies) contain nucleosomal DNA, Ro, La, and the small nuclear ribonucleoproteins. These autoantigen clusters have in common their proximity to the ER and nuclear membranes, sites of increased generation of reactive oxygen species in apoptotic cells. Oxidative modification at these sites may be a mechanism that unites this diverse group of molecules together as autoantigens.

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Selected References

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  1. Arends M. J., Wyllie A. H. Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol. 1991;32:223–254. doi: 10.1016/b978-0-12-364932-4.50010-1. [DOI] [PubMed] [Google Scholar]
  2. Arnett F. C., Olsen M. L., Anderson K. L., Reveille J. D. Molecular analysis of major histocompatibility complex alleles associated with the lupus anticoagulant. J Clin Invest. 1991 May;87(5):1490–1495. doi: 10.1172/JCI115158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bachmann M., Mayet W. J., Schröder H. C., Pfeifer K., Meyer zum Büschenfelde K. H., Müller W. E. Identification of the Ro and La antigens in the endoribonuclease VII--ribonucleoprotein complex. Biochem J. 1987 Apr 1;243(1):189–194. doi: 10.1042/bj2430189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bell D. A., Morrison B., VandenBygaart P. Immunogenic DNA-related factors. Nucleosomes spontaneously released from normal murine lymphoid cells stimulate proliferation and immunoglobulin synthesis of normal mouse lymphocytes. J Clin Invest. 1990 May;85(5):1487–1496. doi: 10.1172/JCI114595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ben-Chetrit E., Chan E. K., Sullivan K. F., Tan E. M. A 52-kD protein is a novel component of the SS-A/Ro antigenic particle. J Exp Med. 1988 May 1;167(5):1560–1571. doi: 10.1084/jem.167.5.1560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boise L. H., González-García M., Postema C. E., Ding L., Lindsten T., Turka L. A., Mao X., Nuñez G., Thompson C. B. bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell. 1993 Aug 27;74(4):597–608. doi: 10.1016/0092-8674(93)90508-n. [DOI] [PubMed] [Google Scholar]
  7. Boveris A., Chance B. The mitochondrial generation of hydrogen peroxide. General properties and effect of hyperbaric oxygen. Biochem J. 1973 Jul;134(3):707–716. doi: 10.1042/bj1340707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burlingame R. W., Rubin R. L., Balderas R. S., Theofilopoulos A. N. Genesis and evolution of antichromatin autoantibodies in murine lupus implicates T-dependent immunization with self antigen. J Clin Invest. 1993 Apr;91(4):1687–1696. doi: 10.1172/JCI116378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Buyon J. P., Slade S. G., Chan E. K., Tan E. M., Winchester R. Effective separation of the 52 kDa SSA/Ro polypeptide from the 48 kDa SSB/La polypeptide by altering conditions of polyacrylamide gel electrophoresis. J Immunol Methods. 1990 May 25;129(2):207–210. doi: 10.1016/0022-1759(90)90440-7. [DOI] [PubMed] [Google Scholar]
  10. Casciola-Rosen L. A., Hubbard A. L. Lumenal labeling of rat hepatocyte early endosomes. Presence of multiple membrane receptors and the Na+,K(+)-ATPase. J Biol Chem. 1992 Apr 25;267(12):8213–8221. [PubMed] [Google Scholar]
  11. Chu J. L., Drappa J., Parnassa A., Elkon K. B. The defect in Fas mRNA expression in MRL/lpr mice is associated with insertion of the retrotransposon, ETn. J Exp Med. 1993 Aug 1;178(2):723–730. doi: 10.1084/jem.178.2.723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cohen J. J., Duke R. C., Fadok V. A., Sellins K. S. Apoptosis and programmed cell death in immunity. Annu Rev Immunol. 1992;10:267–293. doi: 10.1146/annurev.iy.10.040192.001411. [DOI] [PubMed] [Google Scholar]
  13. Cohen P. L., Eisenberg R. A. Lpr and gld: single gene models of systemic autoimmunity and lymphoproliferative disease. Annu Rev Immunol. 1991;9:243–269. doi: 10.1146/annurev.iy.09.040191.001331. [DOI] [PubMed] [Google Scholar]
  14. Cross A. R., Jones O. T. Enzymic mechanisms of superoxide production. Biochim Biophys Acta. 1991 May 6;1057(3):281–298. doi: 10.1016/s0005-2728(05)80140-9. [DOI] [PubMed] [Google Scholar]
  15. Davies K. J. Protein damage and degradation by oxygen radicals. I. general aspects. J Biol Chem. 1987 Jul 15;262(20):9895–9901. [PubMed] [Google Scholar]
  16. 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]
  17. Earnshaw W. C., Rattner J. B. The use of autoantibodies in the study of nuclear and chromosomal organization. Methods Cell Biol. 1991;35:135–175. [PubMed] [Google Scholar]
  18. Eisenberg R. A., Tan E. M., Dixon F. J. Presence of anti-Sm reactivity in autoimmune mouse strains. J Exp Med. 1978 Feb 1;147(2):582–587. doi: 10.1084/jem.147.2.582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Ellis R. E., Yuan J. Y., Horvitz H. R. Mechanisms and functions of cell death. Annu Rev Cell Biol. 1991;7:663–698. doi: 10.1146/annurev.cb.07.110191.003311. [DOI] [PubMed] [Google Scholar]
  21. Fucci L., Oliver C. N., Coon M. J., Stadtman E. R. Inactivation of key metabolic enzymes by mixed-function oxidation reactions: possible implication in protein turnover and ageing. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1521–1525. doi: 10.1073/pnas.80.6.1521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Gammon G., Sercarz E. How some T cells escape tolerance induction. Nature. 1989 Nov 9;342(6246):183–185. doi: 10.1038/342183a0. [DOI] [PubMed] [Google Scholar]
  24. Gavrieli Y., Sherman Y., Ben-Sasson S. A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992 Nov;119(3):493–501. doi: 10.1083/jcb.119.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Gobé G. C., Axelsen R. A., Harmon B. V., Allan D. J. Cell death by apoptosis following X-irradiation of the foetal and neonatal rat kidney. Int J Radiat Biol. 1988 Oct;54(4):567–576. doi: 10.1080/09553008814552011. [DOI] [PubMed] [Google Scholar]
  26. Halliwell B., Gutteridge J. M. Oxygen free radicals and iron in relation to biology and medicine: some problems and concepts. Arch Biochem Biophys. 1986 May 1;246(2):501–514. doi: 10.1016/0003-9861(86)90305-x. [DOI] [PubMed] [Google Scholar]
  27. Hockenbery D. M., Oltvai Z. N., Yin X. M., Milliman C. L., Korsmeyer S. J. Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell. 1993 Oct 22;75(2):241–251. doi: 10.1016/0092-8674(93)80066-n. [DOI] [PubMed] [Google Scholar]
  28. Hopwood D., Coghill G., Ramsay J., Milne G., Kerr M. Microwave fixation: its potential for routine techniques, histochemistry, immunocytochemistry and electron microscopy. Histochem J. 1984 Nov;16(11):1171–1191. doi: 10.1007/BF01003442. [DOI] [PubMed] [Google Scholar]
  29. Huovila A. P., Eder A. M., Fuller S. D. Hepatitis B surface antigen assembles in a post-ER, pre-Golgi compartment. J Cell Biol. 1992 Sep;118(6):1305–1320. doi: 10.1083/jcb.118.6.1305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Hügle B., Hazan R., Scheer U., Franke W. W. Localization of ribosomal protein S1 in the granular component of the interphase nucleolus and its distribution during mitosis. J Cell Biol. 1985 Mar;100(3):873–886. doi: 10.1083/jcb.100.3.873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Itoh N., Yonehara S., Ishii A., Yonehara M., Mizushima S., Sameshima M., Hase A., Seto Y., Nagata S. The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell. 1991 Jul 26;66(2):233–243. doi: 10.1016/0092-8674(91)90614-5. [DOI] [PubMed] [Google Scholar]
  32. Kole R., Fresco L. D., Keene J. D., Cohen P. L., Eisenberg R. A., Andrews P. G. Alu RNA-protein complexes formed in vitro react with a novel lupus autoantibody. J Biol Chem. 1985 Sep 25;260(21):11781–11786. [PubMed] [Google Scholar]
  33. Krainer A. R. Pre-mRNA splicing by complementation with purified human U1, U2, U4/U6 and U5 snRNPs. Nucleic Acids Res. 1988 Oct 25;16(20):9415–9429. doi: 10.1093/nar/16.20.9415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. LeFeber W. P., Norris D. A., Ryan S. R., Huff J. C., Lee L. A., Kubo M., Boyce S. T., Kotzin B. L., Weston W. L. Ultraviolet light induces binding of antibodies to selected nuclear antigens on cultured human keratinocytes. J Clin Invest. 1984 Oct;74(4):1545–1551. doi: 10.1172/JCI111569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lehmann P. V., Forsthuber T., Miller A., Sercarz E. E. Spreading of T-cell autoimmunity to cryptic determinants of an autoantigen. Nature. 1992 Jul 9;358(6382):155–157. doi: 10.1038/358155a0. [DOI] [PubMed] [Google Scholar]
  36. Lerner M. R., Steitz J. A. Antibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5495–5499. doi: 10.1073/pnas.76.11.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Levine B., Huang Q., Isaacs J. T., Reed J. C., Griffin D. E., Hardwick J. M. Conversion of lytic to persistent alphavirus infection by the bcl-2 cellular oncogene. Nature. 1993 Feb 25;361(6414):739–742. doi: 10.1038/361739a0. [DOI] [PubMed] [Google Scholar]
  38. Levine R. L., Oliver C. N., Fulks R. M., Stadtman E. R. Turnover of bacterial glutamine synthetase: oxidative inactivation precedes proteolysis. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2120–2124. doi: 10.1073/pnas.78.4.2120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lin R. H., Mamula M. J., Hardin J. A., Janeway C. A., Jr Induction of autoreactive B cells allows priming of autoreactive T cells. J Exp Med. 1991 Jun 1;173(6):1433–1439. doi: 10.1084/jem.173.6.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Lipham W. J., Redmond T. M., Takahashi H., Berzofsky J. A., Wiggert B., Chader G. J., Gery I. Recognition of peptides that are immunopathogenic but cryptic. Mechanisms that allow lymphocytes sensitized against cryptic peptides to initiate pathogenic autoimmune processes. J Immunol. 1991 Jun 1;146(11):3757–3762. [PubMed] [Google Scholar]
  41. Login G. R., Dvorak A. M. Microwave energy fixation for electron microscopy. Am J Pathol. 1985 Aug;120(2):230–243. [PMC free article] [PubMed] [Google Scholar]
  42. Login G. R., Stavinoha W. B., Dvorak A. M. Ultrafast microwave energy fixation for electron microscopy. J Histochem Cytochem. 1986 Mar;34(3):381–387. doi: 10.1177/34.3.3950387. [DOI] [PubMed] [Google Scholar]
  43. Mamula M. J., Jemmerson R., Hardin J. A. The specificity of human anti-cytochrome c autoantibodies that arise in autoimmune disease. J Immunol. 1990 Mar 1;144(5):1835–1840. [PubMed] [Google Scholar]
  44. Mamula M. J., Lin R. H., Janeway C. A., Jr, Hardin J. A. Breaking T cell tolerance with foreign and self co-immunogens. A study of autoimmune B and T cell epitopes of cytochrome c. J Immunol. 1992 Aug 1;149(3):789–795. [PubMed] [Google Scholar]
  45. Mamula M. J. The inability to process a self-peptide allows autoreactive T cells to escape tolerance. J Exp Med. 1993 Feb 1;177(2):567–571. doi: 10.1084/jem.177.2.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Marti R., Wild P., Schraner E. M., Mueller M., Moor H. Parathyroid ultrastructure after aldehyde fixation, high-pressure freezing, or microwave irradiation. J Histochem Cytochem. 1987 Dec;35(12):1415–1424. doi: 10.1177/35.12.3680934. [DOI] [PubMed] [Google Scholar]
  47. 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]
  48. Mohan C., Adams S., Stanik V., Datta S. K. Nucleosome: a major immunogen for pathogenic autoantibody-inducing T cells of lupus. J Exp Med. 1993 May 1;177(5):1367–1381. doi: 10.1084/jem.177.5.1367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Mosser D. D., Martin L. H. Induced thermotolerance to apoptosis in a human T lymphocyte cell line. J Cell Physiol. 1992 Jun;151(3):561–570. doi: 10.1002/jcp.1041510316. [DOI] [PubMed] [Google Scholar]
  50. Olmsted J. B. Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem. 1981 Dec 10;256(23):11955–11957. [PubMed] [Google Scholar]
  51. Oltvai Z. N., Milliman C. L., Korsmeyer S. J. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell. 1993 Aug 27;74(4):609–619. doi: 10.1016/0092-8674(93)90509-o. [DOI] [PubMed] [Google Scholar]
  52. Pasquali-Ronchetti I., Bini A., Botti B., De Alojsio G., Fornieri C., Vannini V. Ultrastructural and biochemical changes induced by progressive lipid peroxidation on isolated microsomes and rat liver endoplasmic reticulum. Lab Invest. 1980 Apr;42(4):457–468. [PubMed] [Google Scholar]
  53. Rabizadeh S., Oh J., Zhong L. T., Yang J., Bitler C. M., Butcher L. L., Bredesen D. E. Induction of apoptosis by the low-affinity NGF receptor. Science. 1993 Jul 16;261(5119):345–348. doi: 10.1126/science.8332899. [DOI] [PubMed] [Google Scholar]
  54. Reeves W. H., Nigam S. K., Blobel G. Human autoantibodies reactive with the signal-recognition particle. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9507–9511. doi: 10.1073/pnas.83.24.9507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Reveille J. D., Macleod M. J., Whittington K., Arnett F. C. Specific amino acid residues in the second hypervariable region of HLA-DQA1 and DQB1 chain genes promote the Ro (SS-A)/La (SS-B) autoantibody responses. J Immunol. 1991 Jun 1;146(11):3871–3876. [PubMed] [Google Scholar]
  56. Sano H., Morimoto C. Isolation of DNA from DNA/anti-DNA antibody immune complexes in systemic lupus erythematosus. J Immunol. 1981 Feb;126(2):538–539. [PubMed] [Google Scholar]
  57. Sercarz E. E., Lehmann P. V., Ametani A., Benichou G., Miller A., Moudgil K. Dominance and crypticity of T cell antigenic determinants. Annu Rev Immunol. 1993;11:729–766. doi: 10.1146/annurev.iy.11.040193.003501. [DOI] [PubMed] [Google Scholar]
  58. Tan E. M. Autoantibodies in pathology and cell biology. Cell. 1991 Nov 29;67(5):841–842. doi: 10.1016/0092-8674(91)90356-4. [DOI] [PubMed] [Google Scholar]
  59. Tan E. M., Chan E. K., Sullivan K. F., Rubin R. L. Antinuclear antibodies (ANAs): diagnostically specific immune markers and clues toward the understanding of systemic autoimmunity. Clin Immunol Immunopathol. 1988 May;47(2):121–141. doi: 10.1016/0090-1229(88)90066-9. [DOI] [PubMed] [Google Scholar]
  60. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Umansky S. R., Korol' B. A., Nelipovich P. A. In vivo DNA degradation in thymocytes of gamma-irradiated or hydrocortisone-treated rats. Biochim Biophys Acta. 1981 Aug 27;655(1):9–17. doi: 10.1016/0005-2787(81)90060-5. [DOI] [PubMed] [Google Scholar]
  62. Vaux D. L. Toward an understanding of the molecular mechanisms of physiological cell death. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):786–789. doi: 10.1073/pnas.90.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Veis D. J., Sorenson C. M., Shutter J. R., Korsmeyer S. J. Bcl-2-deficient mice demonstrate fulminant lymphoid apoptosis, polycystic kidneys, and hypopigmented hair. Cell. 1993 Oct 22;75(2):229–240. doi: 10.1016/0092-8674(93)80065-m. [DOI] [PubMed] [Google Scholar]
  64. Watanabe-Fukunaga R., Brannan C. I., Copeland N. G., Jenkins N. A., Nagata S. Lymphoproliferation disorder in mice explained by defects in Fas antigen that mediates apoptosis. Nature. 1992 Mar 26;356(6367):314–317. doi: 10.1038/356314a0. [DOI] [PubMed] [Google Scholar]
  65. Wolff S. P., Dean R. T. Fragmentation of proteins by free radicals and its effect on their susceptibility to enzymic hydrolysis. Biochem J. 1986 Mar 1;234(2):399–403. doi: 10.1042/bj2340399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Wolin S. L., Steitz J. A. The Ro small cytoplasmic ribonucleoproteins: identification of the antigenic protein and its binding site on the Ro RNAs. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1996–2000. doi: 10.1073/pnas.81.7.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Wu J., Zhou T., He J., Mountz J. D. Autoimmune disease in mice due to integration of an endogenous retrovirus in an apoptosis gene. J Exp Med. 1993 Aug 1;178(2):461–468. doi: 10.1084/jem.178.2.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  68. Wyllie A. H., Kerr J. F., Currie A. R. Cell death: the significance of apoptosis. Int Rev Cytol. 1980;68:251–306. doi: 10.1016/s0074-7696(08)62312-8. [DOI] [PubMed] [Google Scholar]
  69. Zychlinsky A., Prevost M. C., Sansonetti P. J. Shigella flexneri induces apoptosis in infected macrophages. Nature. 1992 Jul 9;358(6382):167–169. doi: 10.1038/358167a0. [DOI] [PubMed] [Google Scholar]

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