Differentiation of antineutrophil nuclear antibodies in inflammatory bowel and autoimmune liver diseases from antineutrophil cytoplasmic antibodies (p-ANCA) using immunofluorescence microscopy

B. Terjung

Departments of Medicine and of Anatomy and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, USA,

Department of General Internal Medicine I, University of Bonn, Bonn, Germany, and

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H. J. Worman,

H. J. Worman

Departments of Medicine and of Anatomy and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, USA,

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V. Herzog,

V. Herzog

Institute of Cell Biology and Forum Biomedicine, University of Bonn, Ulrich-Bonn, Germany

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T. Sauerbruch,

T. Sauerbruch

Department of General Internal Medicine I, University of Bonn, Bonn, Germany, and

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U. Spengler,

U. Spengler

Department of General Internal Medicine I, University of Bonn, Bonn, Germany, and

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B. Terjung,

B. Terjung

Departments of Medicine and of Anatomy and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, USA,

Department of General Internal Medicine I, University of Bonn, Bonn, Germany, and

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H. J. Worman,

H. J. Worman

Departments of Medicine and of Anatomy and Cell Biology, College of Physicians and Surgeons, Columbia University, New York, USA,

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V. Herzog,

V. Herzog

Institute of Cell Biology and Forum Biomedicine, University of Bonn, Ulrich-Bonn, Germany

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T. Sauerbruch,

T. Sauerbruch

Department of General Internal Medicine I, University of Bonn, Bonn, Germany, and

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U. Spengler,

U. Spengler

Department of General Internal Medicine I, University of Bonn, Bonn, Germany, and

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First published: 07 July 2008

https://doi.org/10.1046/j.1365-2249.2001.01649.x

Citations: 56

Ulrich Spengler, Department of General Internal Medicine I, University of Bonn, Sigmund-Freud-Strasse 25, 53105 Bonn, Germany. E-mail: [email protected]

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Abstract

Perinuclear antineutrophil cytoplasmic antibodies (p-ANCA) directed against cytoplasmic proteins of neutrophils have been studied extensively in patients with systemic vasculitides. Recent data indicate that antineutrophil antibodies in sera from patients with chronic inflammatory bowel diseases (IBD) or autoimmune liver disorders, currently called ‘atypical p-ANCA’, recognize a nuclear target antigen, rendering the term ‘ANCA’ inaccurate. Specific microscopic criteria to distinguish atypical p-ANCA from p-ANCA are lacking. We used planar and confocal laser scanning indirect immunofluorescence microscopy to examine the labelling characteristics of ethanol-, methanol- and formaldehyde-fixed neutrophils by antineutrophil antibodies in 153 serum samples from patients with IBD, autoimmune liver disorders, systemic vasculitides or healthy blood donors. On ethanol- or methanol-fixed neutrophils, multiple intranuclear fluorescent foci together with either a rim-like peripheral nuclear staining (‘type A’) or a combined cytoplasmic and peripheral nuclear staining (‘type B’) was noted exclusively with atypical p-ANCA in sera from patients with IBD or autoimmune liver disorders. Intranuclear foci, which probably corresponded to invaginations of the nuclear envelope, were not labelled by p-ANCA from patients with microscopic polyangiitis or cytoplasmic ANCA (c-ANCA) from patients with Wegener's granulomatosis. On formaldehyde-fixed neutrophils, atypical p-ANCA gave a fine rim-like staining of the nuclear periphery, whereas ANCA diffusely labelled the cytoplasm. To distinguish reliably between the patterns produced by atypical p-ANCA or p-ANCA, particularly p-ANCA, careful indirect immunofluorescence microscopy on ethanol- as well as on formaldehyde-fixed neutrophils is necessary, with particular emphasis on the presence of multiple intranuclear fluorescent foci.

References

1 Woude FJ, Lobatto S, Permin H et al. Autoantibodies against neutrophils and monocytes: tools for diagnosis and marker of disease activity in Wegener's granulomatosis. Lancet 1985; 1: 425–9.
10.1016/S0140-6736(85)91147-X
PubMed Web of Science® Google Scholar
2 Falk RJ & Jennette JC. Anti-neutrophil cytoplasmic autoantibodies with specificity for myeloperoxidase in patients with systemic vasculitis and idiopathic necrotizing and crescentic glomerulonephritis. N Engl J Med 1988; 318: 1651–7.
10.1056/NEJM198806233182504
CAS PubMed Web of Science® Google Scholar
3 Duerr RH, Targan SR, Landers CJ et al. Neutrophil cytoplasmic antibodies: a link between primary sclerosing cholangitis and ulcerative colitis. Gastroenterology 1991; 100: 1385–91.
10.1016/0016-5085(91)70028-V
CAS PubMed Web of Science® Google Scholar
4 Oudkerk-Pool M, Ellerbroek PM, Ridwan BU et al. Serum antineutrophil cytoplasmic autoantibodies in inflammatory bowel disease are mainly associated with ulcerative colitis. A correlation study between perinuclear antineutrophil cytoplasmic autoantibodies and clinical parameters, medical and surgical treatment. Gut 1993; 34: 46–60.
10.1136/gut.34.1.46
PubMed Web of Science® Google Scholar
5 Mulder AHL, Horst G, Haagsma EB, Kleibeuker JH, Kallenberg CGM. Antineutrophil cytoplasmic antibodies in autoimmune liver diseases. Hepatology 1993; 17: 411–7.
10.1002/hep.1840170310
PubMed Web of Science® Google Scholar
6 Hardarson S, LaBrecque DR, Mitros FA, Neil GA, Goeken JA. Antineutrophil cytoplasmic antibody in inflammatory bowel and hepatobiliary diseases. Am J Clin Pathol 1993; 99: 277–81.
10.1093/ajcp/99.3.277
PubMed Web of Science® Google Scholar
7 Targan S, Landers C, Vidrich A, Czaja AL. High-titer antineutrophil cytoplasmic antibodies in type 1 autoimmune hepatitis. Gastroenterology 1995; 108: 1159–66.
10.1016/0016-5085(95)90215-5
PubMed Web of Science® Google Scholar
8 Bansi DS, Fleming KA, Chapman RW. Antineutrophil cytoplasmic antibodies in autoimmune hepatitis. Gastroenterology 1995; 109: 2049–55.
10.1016/0016-5085(95)90785-8
CAS PubMed Web of Science® Google Scholar
9 Zauli D, Ghetti S, Grassi A et al. Anti-neutrophil cytoplasmic antibodies in type 1 and 2 autoimmune hepatitis. Hepatology 1997; 25: 1105–7.
10.1002/hep.510250510
CAS PubMed Web of Science® Google Scholar
10 Walmsley RS, Zhao MH, Hamilton MI et al. Antineutrophil cytoplasm autoantibodies against bactericidal/permeability-increasing protein in inflammatory bowel disease. Gut 1997; 40: 105–9.
10.1136/gut.40.1.105
CAS PubMed Web of Science® Google Scholar
11 Roozendaal C, Zhao MH, Horst G et al. Catalase and α-enolase: two novel granulocyte autoantigens in inflammatory bowel disease (IBD). Clin Exp Immunol 1998; 112: 10–6.DOI: 10.1046/j.1365-2249.1998.00528.x
10.1046/j.1365-2249.1998.00528.x
CAS PubMed Web of Science® Google Scholar
12 Orth T, Kellner R, Diekmann O, Faust J, Meyer zum Büschenfelde KH, Mayet WJ. Identification and characterization of autoantibodies against catalase and α-enolase in patients with primary sclerosing cholangitis. Clin Exp Immunol 1998; 112: 507–15.
10.1046/j.1365-2249.1998.00583.x
CAS PubMed Web of Science® Google Scholar
13 Halbwachs-Mecarelli L, Nusbaum P, Noel LH et al. Antineutrophil cytoplasmic antibodies (ANCA) directed against cathepsin G in ulcerative colitis, Crohn's disease and primary sclerosing cholangitis. Clin Exp Immunol 1992; 90: 79–94.
10.1111/j.1365-2249.1992.tb05835.x
CAS PubMed Web of Science® Google Scholar
14 Peen E, Almer S, Bodemar G et al. Antilactoferrin antibodies and other types of ANCA in ulcerative colitis, primary sclerosing cholangitis and Crohn's disease. Gut 1993; 34: 56–62.
10.1136/gut.34.1.56
CAS PubMed Web of Science® Google Scholar
15 Saxon A, Shanahan F, Landers C, Ganz T, Targan S. A distinct subset of antineutrophil cytoplasmic antibodies is associated with inflammatory bowel disease. J Allergy Clin Immunol 1990; 86: 202–10.
10.1016/S0091-6749(05)80067-3
CAS PubMed Web of Science® Google Scholar
16 Savige J, Gillis D, Benson E et al. International consensus statement on testing and reporting of antineutrophil cytoplasmic antibodies (ANCA). Am J Clin Pathol 1999; 111: 507–13.
10.1093/ajcp/111.4.507
CAS PubMed Web of Science® Google Scholar
17 Rump JA, Schölmerich J, Gross V et al. A new type of perinuclear anti-neutrophil cytoplasmic antibody (p-ANCA) in active ulcerative colitis but not in Crohn's disease. Immunobiology 1990; 181: 406–13.
10.1016/S0171-2985(11)80509-7
CAS PubMed Web of Science® Google Scholar
18 Terjung B, Herzog V, Worman HJ et al. Atypical antineutrophil cytoplasmic antibodies with perinuclear fluorescence in chronic inflammatory bowel diseases and hepatobiliary disorders colocalize with nuclear lamina proteins. Hepatology 1998; 28: 332–40.
10.1002/hep.510280207
CAS PubMed Web of Science® Google Scholar
19 Terjung B, Spengler U, Sauerbruch T, Worman HJ. ‘Atypical p-ANCA’ in IBD and hepatobiliary disorders react with a 50 kD nuclear envelope protein of neutrophils and myeloid cell lines. Gastroenterology 2000; 119: 310–22.
10.1053/gast.2000.9366
CAS PubMed Web of Science® Google Scholar
20 Billing P, Tahir S, Calfin B et al. Nuclear localization of the antigen detected by ulcerative colitis-associated perinuclear antineutrophil cytoplasmic antibodies. Am J Pathol 1995; 147: 979–87.
CAS PubMed Web of Science® Google Scholar
21 Sobajima J, Ozaki S, Uesugi H et al. High mobility group (HMG) non-histone chromosomal proteins HMG1 and HMG2 are significant target antigens of perinuclear antineutrophil cytoplasmic antibodies in autoimmune hepatitis. Gut 1999; 44: 867–73.
10.1136/gut.44.6.867
CAS PubMed Web of Science® Google Scholar
22 Mallolas J, Esteve M, Rius E, Cabré E, Gassull MA. Antineutrophil antibodies associated with ulcerative colitis interact with the antigen(s) during the process of apoptosis. Gut 2000; 47: 74–8.DOI: 10.1136/gut.47.1.74
10.1136/gut.47.1.74
CAS PubMed Web of Science® Google Scholar
23 Eggena M, Cohavy O, Parseghian MP et al. Identification of histone H1 as a cognate antigen of the ulcerative colitis-associated marker antibody p-ANCA. J Autoimmun 2000; 14: 83–97.
10.1006/jaut.1999.0340
CAS PubMed Web of Science® Google Scholar
24 Martin L, Herr JC, Wanamaker W, Kornguth S. Demonstration of specific antineuronal nuclear antibodies in sera of patients with myasthenia gravis. Indirect and direct immunofluorescence. Neurology 1974; 24: 680–3.
10.1212/WNL.24.7.680
CAS PubMed Web of Science® Google Scholar
25 Wiik A. Delineation of a standard procedure for indirect immunofluorescence detection of ANCA. APMIS 1989; 97 (Suppl. 6): 12–3.
Google Scholar
26 Cance WG, Chaudhary N, Worman HJ, Blobel G, Cordon-Cando C. Expression of the nuclear lamins in normal and neoplastic human tissues. J Exp Clin Cancer Res 1992; 11: 233–46.
CAS Web of Science® Google Scholar
27 Spickett GP & Broomhead V. Formalin fixation and patterns of antineutrophil cytoplasmic antibodies. J Clin Pathol 1995; 48: 89–90.
10.1136/jcp.48.1.89
CAS PubMed Web of Science® Google Scholar
28 Fricker M, Hollinshead M, White N, Vaux D. Interphase nuclei of many mammalian cell types contain deep, dynamic, tubular membrane-bound invaginations of the nuclear envelope. J Cell Biol 1997; 136: 531–44.
10.1083/jcb.136.3.531
CAS PubMed Web of Science® Google Scholar
29 Lock RJ. Detection of autoantibodies to neutrophil cytoplasmic antigens. J Clin Pathol 1994; 47: 4–8.
10.1136/jcp.47.1.4
PubMed Web of Science® Google Scholar
30 Mulder AHL, Broekroelofs J, Horst G, Limburg PC, Nelis GF, Kallenberg CGM. Anti-neutrophil cytoplasmic antibodies (ANCA) in inflammatory bowel diseases: characterization and clinical correlates. Clin Exp Immunol 1994; 95: 490–7.
10.1111/j.1365-2249.1994.tb07024.x
CAS PubMed Web of Science® Google Scholar
31 Cambridge G, Rampton DS, Stevens TRJ, McCarthy DA, Kamm M, Leaker B. Antineutrophil antibodies in inflammatory bowel disease: prevalence and diagnostic role. Gut 1992; 33: 668–74.
10.1136/gut.33.5.668
PubMed Web of Science® Google Scholar
32 Chowdhury SM, Broomhead V, Spickett GP, Wilkinson R. Pitfalls of formalin fixation for determination of antineutrophil cytoplasmic antibodies. J Clin Pathol 1999; 52: 475–7.
10.1136/jcp.52.6.475
CAS PubMed Web of Science® Google Scholar
33 Lee SS, Lawton JWM, Chak W. Distinction between antinuclear antibody and p-ANCA. J Clin Pathol 1991; 44: 962–3.
10.1136/jcp.44.11.962
CAS PubMed Web of Science® Google Scholar
34 Bird AG. Is there life in the formalin fixed neutrophil for ANCA testing? No! J Clin Pathol 1999; 52: 403–4.
10.1136/jcp.52.6.403
CAS PubMed Web of Science® Google Scholar

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Volume126, Issue1

October 2001

Pages 37-46

Differentiation of antineutrophil nuclear antibodies in inflammatory bowel and autoimmune liver diseases from antineutrophil cytoplasmic antibodies (p-ANCA) using immunofluorescence microscopy

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Differentiation of antineutrophil nuclear antibodies in inflammatory bowel and autoimmune liver diseases from antineutrophil cytoplasmic antibodies (p-ANCA) using immunofluorescence microscopy

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