Abstract
The objective of this study was to characterize antigens recognized by neutrophil-specific autoantibodies from patients with RA. Sera from 62 RA patients were screened by indirect immunofluorescence (IIF). Positive sera were further tested by ELISAs for antibodies against various granule proteins and by immunoblotting of electrophoretically separated cell, granule or nuclear extracts. Forty-two sera (68%) reacted with ethanol-fixed neutrophils. In the ELISAs 32% of the 28 medium to strongly IIF-positive sera were negative, while 43% were weakly positive for more than one antigen. Immunoblots of whole neutrophils showed IgG reactions at 25–35 kD, in the 55-kD region, at 80 kD, and at 110kD. Most sera reacted with more than one band. Except for the 55-kD antigen, none of the antigens appeared in lymphocytes. The most notable reactivity in subcellular fractions was with lactoferrin and with bands of 25–35 kD from nuclei. In conclusion, anti-neutrophil autoantibodies from RA patients recognize different antigens in the cytoplasm and in the nucleus. Lactoferrin is one of the common antigens recognized, but also unknown nuclear antigens of 25–35 kD mol. wt are involved.
Keywords: rheumatoid arthritis, autoantibodies, neutrophil autoantigens
Full Text
The Full Text of this article is available as a PDF (1.3 MB).
References
- 1.Faber V, Elling P. An antinuclear factor specific for leukocytes. Lancet. 1964:44–5. doi: 10.1016/s0140-6736(64)90283-1. [DOI] [PubMed] [Google Scholar]
- 2.Wiik A. Granulocyte-specific antinuclear antibodies. Possible significance for the pathogenesis, clinical features and diagnosis of rheumatoid arthritis. Allergy. 1980;35:263–89. doi: 10.1111/j.1398-9995.1980.tb01768.x. [DOI] [PubMed] [Google Scholar]
- 3.Davies DJ, Moran JE, Niall JF. Segmental necrotising glomerulonephritis with antineutrophil antibody: possible arbovirus aetiology. Brit Med J. 1982;285:606. doi: 10.1136/bmj.285.6342.606. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Wiik A, van der Woude FJ. The new ACPA/ANCA nomenclature. Neth J Med. 1990;36:107. [PubMed] [Google Scholar]
- 5.Wiik A. Delineation of a standard method for indirect immunofluorescence detection of ANCA. APMIS 97. 1989;(Suppl. 6):12–13. [PubMed] [Google Scholar]
- 6.Wiik A, Rasmussen N, Wieslander J. Methods to detect autoantibodies to neutrophilic granulocytes. In: Maini RN, Van Venrooij WJ, editors. Manual of biological markers of disease. Dordrecht: Kliiwer Academic Publishers; 1993. [Google Scholar]
- 7.Hagen EC, Bailleux BEPB, Es LA, Van Dana MR, van der Woude FJ. Antineutrophil cytoplasmic autoantibodies: a review of the antigens involved, the assays, and the clinical and possible pathogenic consequences. Blood. 1993;81:1996–2002. [PubMed] [Google Scholar]
- 8.Gross WL, Csernok E. Immunodiagnostic and pathophysiologic aspects of antineutrophil cytoplasmic antibodies in vasculitis. Curr Opin Rheumatol. 1995;7:11–19. [PubMed] [Google Scholar]
- 9.Lüdemann J, Utecht B, Gross WL. Anti-neutrophil cytoplasm antibodies in Wegener's granulomatosis recognize an elastinolytic enzyme. J Exp Med. 1990;171:357–62. doi: 10.1084/jem.171.1.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10.Niles JL, McCluskey RT, Ahmad MF, Arnaout AM. Wegener's granulomatosis autoantigen is a novel neutrophil serine proteinase. Blood. 1989;74:1888–93. [PubMed] [Google Scholar]
- 11.Falk RJ, Jennette JC. Anti-neutrophil cytoplasmic autoantibodies with specificity for myeloperoxidase in patients with systemic vasculitis and idiopathic necrotizing and crescentic glomerulonephritis. New Engl J Med. 1988;318:1651–7. doi: 10.1056/NEJM198806233182504. [DOI] [PubMed] [Google Scholar]
- 12.Mulder AHL, Horst G, Van Leeuwen MA, Limburg PC, Kallenberg CGM. Antineutrophil cytoplasmic antibodies in rheumatoid arthritis. Arthritis Rheum. 1993;36:1054–60. doi: 10.1002/art.1780360805. [DOI] [PubMed] [Google Scholar]
- 13.Savige JA, Gallicchio MC, Stockman A, Cunningham TJ, Rowley MJ, Georgiou T, Davies D. Anti-neutrophil cytoplasm antibodies in rheumatoid arthritis. Clin Exp Immunol. 1991;86:92–98. doi: 10.1111/j.1365-2249.1991.tb05779.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14.Bosch X, Llena J, Collado A, Font J, Mirapeix E, Ingelmo M, Munoz-Gómez J, Urbano-Márquez A. Occurrence of antineutrophil cytoplasmic and antineutrophil (peri) nuclear antibodies in rheumatoid arthritis. J Rheumatol. 1995;22:2038–45. [PubMed] [Google Scholar]
- 15.De Bandt M, Meyer O, Haim T, Kahn MF. Antineutrophil cytoplasmic antibodies in rheumatoid arthritis patients. Br J Rheumatol. 1996;35:38–43. doi: 10.1093/rheumatology/35.1.38. [DOI] [PubMed] [Google Scholar]
- 16.Röther E, Schochat T, Peter HH. Antineutrophil cytoplasmic antibodies (ANCA) in rheumatoid arthritis: a prospective study. Rheumatol Int. 1996;15:231–7. doi: 10.1007/BF00290376. [DOI] [PubMed] [Google Scholar]
- 17.Coremans IEM, Hagen EC, Daha MR, van der Woude FJ, van der Keur CK, Breedveld FC. Antilactoferrin antibodies in patients with rheumatoid arthritis are associated with vasculitis. Arthritis Rheum. 1992;35:1466–75. doi: 10.1002/art.1780351210. [DOI] [PubMed] [Google Scholar]
- 18.Schmitt WH, Csemok E, Flesch BK, Hauschild S, Gross WL. Autoantibodies directed against lysozyme: a new target antigen for anti-neutrophil cytoplasmatic antibodies (ANCA) In: Gross WL, editor. ANCA-associated vasculitides: immunological and clinical aspects. New York: Plenum Press; 1993. pp. 267–71. [Google Scholar]
- 19.Lesavre P, Noel LH, Gayno S, Nusbaum P, Remeaux D, Erlinger S, Grünfeld JP, Halbwachs-Mecarelli L. Atypical autoantigen targets of perinuclear antineutrophil cytoplasm antibodies (P-ANCA): specificity and clinical associations. J Autoimmun. 1993;6:185–95. doi: 10.1006/jaut.1993.1016. [DOI] [PubMed] [Google Scholar]
- 20.Cambridge G, Williams M, Leaker B, Corbett M, Smith CR. Antimyeloperoxidase antibodies in patients with rheumatoid arthritis: prevalence, clinical correlates and IgG subclass. Ann Rheum Dis. 1994;53:24–29. doi: 10.1136/ard.53.1.24. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21.Amett FC, Edworthy SM, Bloch DA, et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 1988;31:315–24. doi: 10.1002/art.1780310302. [DOI] [PubMed] [Google Scholar]
- 22.LaCour BB, Wiik A, Hoier-Madsen M, Rasmussen N, Baslund B. Clinical correlates and substrate specificities of antibodies exhibiting neutrophilocyte nuclear reactivity. J Immunol Methods. 1995;187:287–95. doi: 10.1016/0022-1759(95)00196-7. [DOI] [PubMed] [Google Scholar]
- 23.Hagen EC, Andrassy K, Csernok E, et al. Development of solid phase assays for the detection of anti-neutrophil cytoplasmic antibodies (ANCA). A report on the second phase of an international cooperative study on the standardization of ANCA assays. J. Immunol Methods. 1996;196:1–15. doi: 10.1016/0022-1759(96)00111-1. [DOI] [PubMed] [Google Scholar]
- 24.Borregaard N, Heiple JM, Simons ER, Clark RA. Subcellular localization of the b-cytochrome component of the human neutrophil microbicidal oxidase: translocation during activation. J Cell Biol. 1983;97:52–61. doi: 10.1083/jcb.97.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 25.Mulder AHL, Horst G, Haagsma EB, Limburg PC, Kleibeuker JH, Kallenberg CGM. Prevalence and characterization of neutrophil cytoplasmic antibodies in autoimmune liver diseases. Hepatology. 1993;17:411–7. [PubMed] [Google Scholar]
- 26.Larsson LI. Immunocytochemistry: theory and practice. Boca Raton: CRC Press; 1988. pp. 41–76. [Google Scholar]
- 27.Spickett GP, Broomhead V. Formalin fixation and patterns of antineutrophil cytoplasmic antibodies. Sarcoidosis, vasculitis and diffuse lung diseases. 1996;13:265. doi: 10.1136/jcp.48.1.89. Abstr. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 28.Wiik A. Anti-neutrophil cytoplasmic antibodies in primary small vessel vasculitides. Scand J Rheumatol. 1996;25:65–69. doi: 10.3109/03009749609069209. [DOI] [PubMed] [Google Scholar]
- 29.Hassfeld W, Steiner G, Hartmut K. Demonstration of a new antinuclear antibody (anti-RA33) that is highly specific for rheumatoid arthritis. Arthritis Rheum. 1989;32:1515–20. doi: 10.1002/anr.1780321204. [DOI] [PubMed] [Google Scholar]
- 30.Zhao MH, Jones SJ, Lockwood CM. Bactericidal/permeability-increasing protein (BPI) is an important antigen for anti-neutrophil cytoplasmic antibodies (ANCA) in vasculitis. Clin Exp Immunol. 1995;99:49–56. doi: 10.1111/j.1365-2249.1995.tb03471.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
