AN ELISA FOR THE DETECTION OF ANTI-NEUTROPHIL CYTOPLASMIC ANTIBODIES USING THE CYTOPLASMIC α-FRACTION -CLINICAL USE FOR THE DIAGNOSIS OF WEGENER GRANULOMATOSIS AND FOR MONITORING ITS ACTIVITY
Anti-neutrophil cytoplasmic antibodies(ANCA)have been reported to be a disease-specific marker for Wegener's granulomatosis(WG). In the present study we developed an enzyme-linked immunosorbent assay(ELISA)for detecting and quantifying ANCA. The cytoplasmic α-fraction of neutrophils obtained fr...
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| Published in | Nippon Jibi Inkoka Gakkai Kaiho Vol. 97; no. 1; pp. 20 - 27 |
|---|---|
| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English Japanese |
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Japan
The Oto-Rhino-Laryngological Society of Japan, Inc
1994
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| ISSN | 0030-6622 1883-0854 |
| DOI | 10.3950/jibiinkoka.97.20 |
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| Abstract | Anti-neutrophil cytoplasmic antibodies(ANCA)have been reported to be a disease-specific marker for Wegener's granulomatosis(WG). In the present study we developed an enzyme-linked immunosorbent assay(ELISA)for detecting and quantifying ANCA. The cytoplasmic α-fraction of neutrophils obtained from healthy human donors was used as an antigen. The α-fraction was purified from supernatants of homogenized neutrophils by sucrose gradient centrifugation. Peroxidase conjugated rabbit anti-human IgG was used as a secondary antibody. Diluted sera from 13 patients with WG, 15 with Sjogren syndrome, 1 with polymorphic reticulosis, 3 with relapsing polychondritis, 4 with other collagen diseases, 4 with sinusitis and 18 healthy donors were examined. It was concluded that this ELISA was sufficiently specific and sensitive for WG, and the ELISA units correlated individually with the ANCA titers as determined by an indirect immunofluorescence technique. This ELISA provides precise ANCA quantitation and will be useful for the diagnosis of WG and for monitoring its activity. |
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| AbstractList | Anti-neutrophil cytoplasmic antibodies(ANCA)have been reported to be a disease-specific marker for Wegener's granulomatosis(WG). In the present study we developed an enzyme-linked immunosorbent assay(ELISA)for detecting and quantifying ANCA. The cytoplasmic α-fraction of neutrophils obtained from healthy human donors was used as an antigen. The α-fraction was purified from supernatants of homogenized neutrophils by sucrose gradient centrifugation. Peroxidase conjugated rabbit anti-human IgG was used as a secondary antibody. Diluted sera from 13 patients with WG, 15 with Sjogren syndrome, 1 with polymorphic reticulosis, 3 with relapsing polychondritis, 4 with other collagen diseases, 4 with sinusitis and 18 healthy donors were examined. It was concluded that this ELISA was sufficiently specific and sensitive for WG, and the ELISA units correlated individually with the ANCA titers as determined by an indirect immunofluorescence technique. This ELISA provides precise ANCA quantitation and will be useful for the diagnosis of WG and for monitoring its activity. Anti-neutrophil cytoplasmic antibodies (ANCA) have been reported to be a disease-specific marker for Wegener's granulomatosis (WG). In the present study we developed an enzyme-linked immunosorbent assay (ELISA) for detecting and quantifying ANCA. The cytoplasmic alpha-fraction of neutrophils obtained from healthy human donors was used as an antigen. The alpha-fraction was purified from supernatants of homogenized neutrophils by sucrose gradient centrifugation. Peroxidase conjugated rabbit anti-human IgG was used as a secondary antibody. Diluted sera from 13 patients with WG, 15 with Sjögren syndrome, 1 with polymorphic reticulosis, 3 with relapsing polychondritis, 4 with other collagen diseases, 4 with sinusitis and 18 healthy donors were examined. It was concluded that this ELISA was sufficiently specific and sensitive for WG, and the ELISA units correlated individually with the ANCA titers as determined by an indirect immunofluorescence technique. This ELISA provides precise ANCA quantitation and will be useful for the diagnosis of WG and for monitoring its activity. Anti-neutrophil cytoplasmic antibodies (ANCA) have been reported to be a disease-specific marker for Wegener's granulomatosis (WG). In the present study we developed an enzyme-linked immunosorbent assay (ELISA) for detecting and quantifying ANCA. The cytoplasmic alpha-fraction of neutrophils obtained from healthy human donors was used as an antigen. The alpha-fraction was purified from supernatants of homogenized neutrophils by sucrose gradient centrifugation. Peroxidase conjugated rabbit anti-human IgG was used as a secondary antibody. Diluted sera from 13 patients with WG, 15 with Sjögren syndrome, 1 with polymorphic reticulosis, 3 with relapsing polychondritis, 4 with other collagen diseases, 4 with sinusitis and 18 healthy donors were examined. It was concluded that this ELISA was sufficiently specific and sensitive for WG, and the ELISA units correlated individually with the ANCA titers as determined by an indirect immunofluorescence technique. This ELISA provides precise ANCA quantitation and will be useful for the diagnosis of WG and for monitoring its activity.Anti-neutrophil cytoplasmic antibodies (ANCA) have been reported to be a disease-specific marker for Wegener's granulomatosis (WG). In the present study we developed an enzyme-linked immunosorbent assay (ELISA) for detecting and quantifying ANCA. The cytoplasmic alpha-fraction of neutrophils obtained from healthy human donors was used as an antigen. The alpha-fraction was purified from supernatants of homogenized neutrophils by sucrose gradient centrifugation. Peroxidase conjugated rabbit anti-human IgG was used as a secondary antibody. Diluted sera from 13 patients with WG, 15 with Sjögren syndrome, 1 with polymorphic reticulosis, 3 with relapsing polychondritis, 4 with other collagen diseases, 4 with sinusitis and 18 healthy donors were examined. It was concluded that this ELISA was sufficiently specific and sensitive for WG, and the ELISA units correlated individually with the ANCA titers as determined by an indirect immunofluorescence technique. This ELISA provides precise ANCA quantitation and will be useful for the diagnosis of WG and for monitoring its activity. |
| Author | RYU, TAKUSYU FUKUDA, SATOSHI ORIDATE, NOBUHIKO HONKE, KOICHI MAGUCHI, SHIRO TAKIZAWA, MASAHIKO INUYAMA, YUKIO |
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| References | 25) Falk RJ, Terrell R, Jennette JC et al: Antineutro-phil cytoplasmic antibodies (ANCA) stimulate neutrophil activation in vitro. Kidney Int 35: 346, 1989. 5) Lüdemann J, Utecht B, Gross WL: Anti-neutrophil cytoplasm antibodies in Wegener's granulomatosis recognize an elastinolytic enzyme. J Exp Med 171: 357-362, 1990. 26) Katz P, Ailing DW, Haynes BP et al: Association of Wegener's granulomatosis with HLA-B8. Clin Immunopathol 14: 268-271, 1979. 12) Goldschmeding R, van der Schoot CE, ten Bokkel Huinink D et al: Wegener's granulomatosis autoantibodies identify a novel diisopropylfluor-ophosphate-binding protein in the lysosomes of normal human neutrophils. J Clin Invest 84, 1989. 16) 滝沢昌彦,間口一郎,折舘伸彦,福田諭,犬山征夫他:抗ヒト好中球細胞質抗体の推移を追えたWegener肉芽腫症の1例.耳鼻と臨床 39:422-428,1993. 20) Rasmussen N, Lüdemann J, Utecht B: ELISA examination for IgG-ANCA in sera submitted for the 1st international workshop on ANCA. APMIS 97 (Suppl 6): 21-22, 1989. 13) Gross WL, Csernok E, Schmitt WH et al: Antineutrophil cytoplasmic autoantibodies: immunobiological aspects. Klin Wochenschr 69: 558 -566, 1991. 2) Lüidemann J, Gross WL. Autoantibodies against cytoplasmic structures of neutrophil granulocytes in Wegener's granulomatosis. Clin Exp Immunol 69: 350-357, 1987. 6) Brown WJ, Shannon WA, Snell WJ: Specific and azurophilic granules from rabbit polymorphonuclear leukocytes. J Cell Biology 96: 1030-1039, 1983. 17) Savage COS, Winearls CG, Jones S et al: Prospective study of radioimmunoassay for antibodies against neutrophil cytoplasm in diagnosis of systemic vasculitis. Lancet ii: 1389-1393, 1987. 15) 間口四郎,滝沢昌彦,劉沢周,福田諭,犬山征夫他:Wegener肉芽腫症早期診断のための抗好中球細胞質抗体(ANCA)測定の重要性.日耳算 96:1288-1296,1993. 11) Falk RJ, Jennette JC: Wegener's granulomatosis, systemic vasculitis, and antineutrophil cytoplasmic autoantibodies. Annu Rev Med 42: 459-469, 1991. 7) Fauci AS, Hynes BF, Katz P et al: Wegener's granulomatosis: prospective clinical and therapeutic experience with 85 patients for 21 years. Ann Intern Med 98: 76-85, 1983. 9) 犬山征夫,福田諭,熊谷雅彦:ウェゲナー肉芽腫症の耳鼻咽喉科的病変.リウマチ科2:623-629,1989. 8) 近藤信雄,吉木敬:ウェゲナー肉芽腫症の病現所見.リウマチ科2:649-656,1989. 18) Lüdemann J, Utecht B, Gross WL: Detection and quantitation of anti-neutrophil cytoplasm antibodies in Wegener's granulomatosis by ELISA using affinity-purified antigen. J Immunol Methods 114: 167-174, 1988. 24) Keogan MT, Esnault VLM, Lockwood CM et al: Activation of normal neutrophils by anti-neutrophil cytoplasm antibodies. Clin exp Immunol 90: 228-234,1992. 27) Elkon KB, Sutherland DC, Rees AJ: Brief report: HLA antigen frequencies in systemic vasculitis: increase in HLA - DR2 in Wegener's Granulomatosis. Arthritis Reum 26: 102-103, 1983. 14) 吉田雅治:ウェゲナー肉芽腫症と自己抗体,リウマチ科2:659-662,1989. 3) Specks U, Christine L, Wheatley BA: Anticytoplas. mic autoantibodies in the diagnosis and follow-up of Wegener's granulomatosis. Mayo Clin Proc. 64 : 28-36. 1989. 10) 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 318, 1988. 4) Cohen Tervaert JW, Huitema M, Hene RJ et al: Prevention of relapses in Wegener's granulomatosis by treatment based on antineutrophil cytoplasmic antibody titer. Lancet 336: 709-711, 1990. 1) van der Woude FJ, Rasmussen N, Lobatto S et al: Autoantibodies against neutrophils and monocytes: tool for diagnosis and marker of disease activity in Wegener's granulomatosis. Lancet i: 425-429, 1985. 23) Carnok E, Lüdemann J, Gross WL et al: Ultrastructual localization of proteinase 3, the target antigen of anti -cytoplasmic antibodies circulating in Wegener's granulomatosis. Am J Pathol 137: 1113, 1990. 19) Rasmusses N, Borregaard N, Wieslander J et al: Alpha-ELISA determination of ANCA and characterization of the ANCA-related antigen. APMIS 97 (suppl 6): 40, 1989. 21) Rasmussen N, Sjölin C, Isaksson B et al: An ELISA for the detection of anti neutrophil cytoplasm antibodies (ANCA). J Immunol Methods 127: 139-145, 1990. 22) Yoshida M, Nagasawa T: Clinical significance of anti-neutrophil cytoplasmic antibodies (ANCA) in collagen diseases associated with vasculitic syndrome in Japan. Japa J Nephrology 34: 371-378, 1992. |
| References_xml | – reference: 5) Lüdemann J, Utecht B, Gross WL: Anti-neutrophil cytoplasm antibodies in Wegener's granulomatosis recognize an elastinolytic enzyme. J Exp Med 171: 357-362, 1990. – reference: 3) Specks U, Christine L, Wheatley BA: Anticytoplas. mic autoantibodies in the diagnosis and follow-up of Wegener's granulomatosis. Mayo Clin Proc. 64 : 28-36. 1989. – reference: 8) 近藤信雄,吉木敬:ウェゲナー肉芽腫症の病現所見.リウマチ科2:649-656,1989. – reference: 9) 犬山征夫,福田諭,熊谷雅彦:ウェゲナー肉芽腫症の耳鼻咽喉科的病変.リウマチ科2:623-629,1989. – reference: 1) van der Woude FJ, Rasmussen N, Lobatto S et al: Autoantibodies against neutrophils and monocytes: tool for diagnosis and marker of disease activity in Wegener's granulomatosis. Lancet i: 425-429, 1985. – reference: 11) Falk RJ, Jennette JC: Wegener's granulomatosis, systemic vasculitis, and antineutrophil cytoplasmic autoantibodies. Annu Rev Med 42: 459-469, 1991. – reference: 17) Savage COS, Winearls CG, Jones S et al: Prospective study of radioimmunoassay for antibodies against neutrophil cytoplasm in diagnosis of systemic vasculitis. Lancet ii: 1389-1393, 1987. – reference: 13) Gross WL, Csernok E, Schmitt WH et al: Antineutrophil cytoplasmic autoantibodies: immunobiological aspects. Klin Wochenschr 69: 558 -566, 1991. – reference: 21) Rasmussen N, Sjölin C, Isaksson B et al: An ELISA for the detection of anti neutrophil cytoplasm antibodies (ANCA). J Immunol Methods 127: 139-145, 1990. – reference: 19) Rasmusses N, Borregaard N, Wieslander J et al: Alpha-ELISA determination of ANCA and characterization of the ANCA-related antigen. APMIS 97 (suppl 6): 40, 1989. – reference: 24) Keogan MT, Esnault VLM, Lockwood CM et al: Activation of normal neutrophils by anti-neutrophil cytoplasm antibodies. Clin exp Immunol 90: 228-234,1992. – reference: 10) 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 318, 1988. – reference: 20) Rasmussen N, Lüdemann J, Utecht B: ELISA examination for IgG-ANCA in sera submitted for the 1st international workshop on ANCA. APMIS 97 (Suppl 6): 21-22, 1989. – reference: 6) Brown WJ, Shannon WA, Snell WJ: Specific and azurophilic granules from rabbit polymorphonuclear leukocytes. J Cell Biology 96: 1030-1039, 1983. – reference: 18) Lüdemann J, Utecht B, Gross WL: Detection and quantitation of anti-neutrophil cytoplasm antibodies in Wegener's granulomatosis by ELISA using affinity-purified antigen. J Immunol Methods 114: 167-174, 1988. – reference: 27) Elkon KB, Sutherland DC, Rees AJ: Brief report: HLA antigen frequencies in systemic vasculitis: increase in HLA - DR2 in Wegener's Granulomatosis. Arthritis Reum 26: 102-103, 1983. – reference: 16) 滝沢昌彦,間口一郎,折舘伸彦,福田諭,犬山征夫他:抗ヒト好中球細胞質抗体の推移を追えたWegener肉芽腫症の1例.耳鼻と臨床 39:422-428,1993. – reference: 12) Goldschmeding R, van der Schoot CE, ten Bokkel Huinink D et al: Wegener's granulomatosis autoantibodies identify a novel diisopropylfluor-ophosphate-binding protein in the lysosomes of normal human neutrophils. J Clin Invest 84, 1989. – reference: 2) Lüidemann J, Gross WL. Autoantibodies against cytoplasmic structures of neutrophil granulocytes in Wegener's granulomatosis. Clin Exp Immunol 69: 350-357, 1987. – reference: 7) Fauci AS, Hynes BF, Katz P et al: Wegener's granulomatosis: prospective clinical and therapeutic experience with 85 patients for 21 years. Ann Intern Med 98: 76-85, 1983. – reference: 23) Carnok E, Lüdemann J, Gross WL et al: Ultrastructual localization of proteinase 3, the target antigen of anti -cytoplasmic antibodies circulating in Wegener's granulomatosis. Am J Pathol 137: 1113, 1990. – reference: 15) 間口四郎,滝沢昌彦,劉沢周,福田諭,犬山征夫他:Wegener肉芽腫症早期診断のための抗好中球細胞質抗体(ANCA)測定の重要性.日耳算 96:1288-1296,1993. – reference: 14) 吉田雅治:ウェゲナー肉芽腫症と自己抗体,リウマチ科2:659-662,1989. – reference: 4) Cohen Tervaert JW, Huitema M, Hene RJ et al: Prevention of relapses in Wegener's granulomatosis by treatment based on antineutrophil cytoplasmic antibody titer. Lancet 336: 709-711, 1990. – reference: 22) Yoshida M, Nagasawa T: Clinical significance of anti-neutrophil cytoplasmic antibodies (ANCA) in collagen diseases associated with vasculitic syndrome in Japan. Japa J Nephrology 34: 371-378, 1992. – reference: 26) Katz P, Ailing DW, Haynes BP et al: Association of Wegener's granulomatosis with HLA-B8. Clin Immunopathol 14: 268-271, 1979. – reference: 25) Falk RJ, Terrell R, Jennette JC et al: Antineutro-phil cytoplasmic antibodies (ANCA) stimulate neutrophil activation in vitro. Kidney Int 35: 346, 1989. |
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| Snippet | Anti-neutrophil cytoplasmic antibodies(ANCA)have been reported to be a disease-specific marker for Wegener's granulomatosis(WG). In the present study we... Anti-neutrophil cytoplasmic antibodies (ANCA) have been reported to be a disease-specific marker for Wegener's granulomatosis (WG). In the present study we... |
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| SubjectTerms | anti-neutrophil cytoplasmic antibody (ANCA) ELISA Antibodies, Antineutrophil Cytoplasmic Autoantibodies - analysis Biomarkers - analysis Cell Fractionation Cytoplasm - immunology Enzyme-Linked Immunosorbent Assay Granulomatosis with Polyangiitis - diagnosis Granulomatosis with Polyangiitis - immunology Humans Neutrophils - immunology Predictive Value of Tests Sensitivity and Specificity α-fraction |
| Title | AN ELISA FOR THE DETECTION OF ANTI-NEUTROPHIL CYTOPLASMIC ANTIBODIES USING THE CYTOPLASMIC α-FRACTION -CLINICAL USE FOR THE DIAGNOSIS OF WEGENER GRANULOMATOSIS AND FOR MONITORING ITS ACTIVITY |
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