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Chapter 118 : Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides

Author: Allan Wiik
Content Type: Reference
Publication Year: 2006

Category: Immunology

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Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides, Page 1 of 2

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Abstract:

This chapter concentrates on vasculitides which are associated with presence of antineutrophil cytoplasm antibodies (ANCAs), it focuses on idiopathic smallvessel vasculitides (SVV), and mentions some of the clinical conditions where other types of neutrophil-specific autoantibodies (NSA) are commonly produced. The diagnosis of a primary vasculitic condition must rest on sound clinical judgment of more or less characteristic constellations of symptoms and features, with some being indicative of vasculitis and others reflecting just a general inflammatory condition (arthralgias, fever, fatigue, loss of appetite, hypersedimentation). Although the chapter focuses on the methodologies used to detect ANCAs, it also stresses important aspects of setting clinically validated assay cutoff values, controlling laboratory performance quality, knowing frequent pitfalls, and how to exercise troubleshooting. It also mentions possibilities for reporting laboratory data with due consideration toward differential diagnostics, potential clinical utility, and interpretation. Monoclonal mouse antibodies to proteinase 3 (PR3), myeloperoxidase (MPO), and elastase (EL) have been used to capture the respective antigens from cell extracts, and the captured antigen has then been used as the target in an enzyme-linked immunosorbent assay (ELISA).

Citation: Wiik A. 2006. Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides, p 1053-1058. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch118

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Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides
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Citation: Wiik A. 2006. Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides, p 1053-1058. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch118
/content/10.1128/9781555815905.ch118.ch118fig01
FIGURE 1
Typical C-ANCA pattern seen on ethanol-fixed neutrophils and monocytes by PR3-ANCA-containing sera, here stained for IgG.
10.1128/9781555815905/f1054-01_thmb.gif
10.1128/9781555815905/f1054-01.gif
Image of FIGURE 1
FIGURE 1

Typical C-ANCA pattern seen on ethanol-fixed neutrophils and monocytes by PR3-ANCA-containing sera, here stained for IgG.

Citation: Wiik A. 2006. Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides, p 1053-1058. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch118
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/content/10.1128/9781555815905.ch118.ch118fig02
FIGURE 2
Typical P-ANCA pattern seen on ethanol-fixed neutrophils by MPO-ANCA-containing sera, here stained for IgG.
10.1128/9781555815905/f1054-02_thmb.gif
10.1128/9781555815905/f1054-02.gif
Image of FIGURE 2
FIGURE 2

Typical P-ANCA pattern seen on ethanol-fixed neutrophils by MPO-ANCA-containing sera, here stained for IgG.

Citation: Wiik A. 2006. Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides, p 1053-1058. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch118
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/content/10.1128/9781555815905.ch118.ch118fig03
FIGURE 3
Diagram showing typical peripheral staining of neutrophil (PMN) and monocyte (MO) nuclei and the nucleus of an adjacent lymphocyte (LY), while a more distantly located lymphocyte and an eosinophil (EO) are not stained. The artifac-tual ANA-like pattern on positive cells is due to redistribution of soluble, diffusible molecules (such as the cationic MPO) onto the anionic nuclei located close by, here symbolized by the circles around the PMN and monocyte.
10.1128/9781555815905/f1056-01_thmb.gif
10.1128/9781555815905/f1056-01.gif
Image of FIGURE 3
FIGURE 3

Diagram showing typical peripheral staining of neutrophil (PMN) and monocyte (MO) nuclei and the nucleus of an adjacent lymphocyte (LY), while a more distantly located lymphocyte and an eosinophil (EO) are not stained. The artifac-tual ANA-like pattern on positive cells is due to redistribution of soluble, diffusible molecules (such as the cationic MPO) onto the anionic nuclei located close by, here symbolized by the circles around the PMN and monocyte.

Citation: Wiik A. 2006. Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides, p 1053-1058. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch118
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/content/10.1128/9781555815905.ch118.ch118fig04
FIGURE 4
Diagram showing typical peripheral staining of all leukocyte nuclei by serum containing anti-double-stranded DNA. Diffusion of soluble and freely diffusible molecules such as MPO is symbolized by the circles around the PMN and the monocyte (MO). LY, lymphocyte; EO, eosinophil. Note that all cell nuclei in the field are positive.
10.1128/9781555815905/f1056-02_thmb.gif
10.1128/9781555815905/f1056-02.gif
Image of FIGURE 4
FIGURE 4

Diagram showing typical peripheral staining of all leukocyte nuclei by serum containing anti-double-stranded DNA. Diffusion of soluble and freely diffusible molecules such as MPO is symbolized by the circles around the PMN and the monocyte (MO). LY, lymphocyte; EO, eosinophil. Note that all cell nuclei in the field are positive.

Citation: Wiik A. 2006. Antineutrophil Cytoplasm Antibodies (ANCAs) and Strategy for Diagnosing ANCA-Associated Vasculitides, p 1053-1058. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch118
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References

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1. Goldschmeding, R.,, E. C. van der Schoot,, D. Ten Bokkel Huinink,, C. E. Hack,, M. E. van den Ende,, C. G. M. Kallenberg, and , A. E. G. K. von dem Borne. 1989. Wegener’s granulomatosis autoantibodies identify a novel diisopropyl-fluorophosphate binding protein in the lyso-somes of normal human neutrophils. J. Clin. Investig. 84: 1577– 1587.
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5. Jennette, J. C.,, R. J. Falk,, K. Andrassy,, P. A. Bacon,, J. Churg,, W L. Gross,, E. C. Hagen,, G. S. Hoffman,, G. G. Hunder,, C. G. M. Kallenberg,, R. T. McCluskey,, R. A. Sinico,, A. J. Rees,, L. A. van Es,, R. Waldherr, and , A. Wiik. 1994. Nomenclature of systemic vasculitides: the proposal of an international consensus conference. Arthritis Rheum. 37: 187– 192.
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8. Savige, J.,, D. Gillis,, E. Benson,, D. Davies,, V. Esnault,, R. J. Falk,, E. C. Hagen,, D. Jayne,, J. C. Jennette,, B. Paspaliaris,, W Pollock,, C. Pusey,, C. O. Savage,, R. Silvestrini,, E. van der Woude,, J. Wieslander, and A. Wiik for the International Group for Consensus Statement on Testing and Reporting of Antineutrophil Cytoplasmic Antibodies. 1999. International consensus statement on testing and reporting of antineutrophil cytoplasmic antibodies (ANCA). Am. J. Clin. Pathol. 111: 507– 513.
9. Segelmark, M.,, B. Baslund, and , J. Wieslander. 1994. Some patients with anti-myeloperoxidase antibodies have a c-ANCA pattern. Clin. Exp. Immunol. 96: 458– 465.
10. Specks, D.,, D. N. Fass,, M. P. Fautsch,, A. M. Himmel, and , M. A. Viss. 1996. Recombinant human proteinase 3, the Wegener’s autoantigen, expressed in HMC-l cells is enzymatically active and recognized by c-ANCA. FEBS Lett. 390: 265– 270.
11. Sun, J.,, D. N. Fass,, J. A. Hudson,, M. A. Viss,, J. Wieslander,, H. A. Homburger, and , D. Specks. 1998. Capture-ELISA based on recombinant proteinase 3 (PR3) is sensitive for PR3-ANCA testing and allows detection of PR3 and PR3-ANCA/PR3 immune complexes. J. Immunol. Methods 211: 111– 123.
12. Sun, J.,, D. N. Fass,, M. A. Viss,, A. M. Hummel,, H. Tang,, H. A. Homburger, and , D. Specks. 1998. A proportion of proteinase 3-specific anti-neutrophil cytoplasmic antibodies only react with proteinase 3 after cleavage of its N-terminal activation peptide. Clin. Exp. Immunol. 114: 320– 326.
13. Westman, K. W. A.,, D. Selga,, P. Bygren,, M. Segelmark,, B. Baslund,, A. Wiik, and , J. Wieslander. 1998. Clinical evaluation of a capture ELISA for detection of proteinase 3 antineutrophil cytoplasmic antibody. Kidney Int. 53: 1230– 1236.
14. Wiik, A. 1989. Delineation of a standard procedure for indirect immunofluorescence detection of ANCA. Acta Pathol. Microbiol. Immunol. Scand. 97(Suppl. 6): 12– 13.

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