A Dual-Fixed Neutrophil Substrate Improves Interpretation of Antineutrophil Cytoplasmic Antibodies by Indirect Immunofluorescence

Lin, Ming‐Wei; Silvestrini, Roger; Culican, Suzanne; Campbell, D. S.; Fulcher, David A.

doi:10.1309/ajcpg02fgqvausiu

Cited by 12 publications

(6 citation statements)

References 19 publications

(22 reference statements)

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“…On the other hand, although limited, the misinterpretation of A-ANCA, as P-ANCA, cannot be excluded since our detection was based on immunofluorescence detection of only ethanol-fixed neutrophils. The parallel use of ethanol and formaldehyde-fixed neutrophils for the detection of P-ANCA has been reported to discriminate false P-, atypical ANCA [33]. However, this approach applies mainly for recognizing P-ANCA targeting MPO, whereas it has been reported to be non-beneficial for the study of SARDs patients other than those with vasculitides [33][34][35].…”

Section: Discussionmentioning

confidence: 99%

“…The parallel use of ethanol and formaldehyde-fixed neutrophils for the detection of P-ANCA has been reported to discriminate false P-, atypical ANCA [33]. However, this approach applies mainly for recognizing P-ANCA targeting MPO, whereas it has been reported to be non-beneficial for the study of SARDs patients other than those with vasculitides [33][34][35]. Furthermore, the likelihood of the detection of P-ANCA-related pattern in the setting of concurrent high ANA titers cannot be excluded as ANA may produce indistinguishable immunofluorescent staining patterns on ethanol-fixed neutrophils.…”

Section: Discussionmentioning

confidence: 99%

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Occurrence and Antigenic Specificity of Perinuclear Anti-Neutrophil Cytoplasmic Antibodies (P-ANCA) in Systemic Autoimmune Diseases

Argyropoulou

Goules

Boutzios

et al. 2021

Cells

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Perinuclear anti-neutrophilic cytoplasmic antibodies (P-ANCA) recognize heterogeneous antigens, including myeloperoxidase (MPO), lactoferrin, elastase, cathepsin-G and bactericidal/permeability-increasing protein. Although P-ANCA have diagnostic utility in vasculitides, they may also be found in patients with various other systemic autoimmune rheumatic diseases (SARDs). Nevertheless, the clinical significance and the targets recognized by P-ANCA in such patients remain unclear. For this purpose, herein we investigated the occurrence of ANCA-related antigenic specificities in 82 P-ANCA-positive sera by multiplex ELISA, as well as their association with other autoantibodies. The P-ANCA-positive sera corresponded to patients with vasculitides (n = 24), systemic lupus erythematosus (n = 28), antiphospholipid syndrome (n = 5), Sjögren’s syndrome (n = 7), rheumatoid arthritis (n = 3), systemic scleroderma (n = 1), sarcoidosis (n = 1) and Hashimoto′s thyroiditis (n = 13). In most P-ANCA-positive patients studied (51/82, 62.3%), these autoantibodies occurred in high titers (>1:160). The analysis of P-ANCA-positive sera revealed reactivity to MPO in only 50% of patients with vasculitides, whereas it was infrequent in the other disease groups studied. Reactivity to other P-ANCA-related autoantigens was also rarely detected. Our findings support that high P-ANCA titers occur in SARD. The P-ANCA-positive staining pattern is associated with MPO specificity in vasculitides, while in other autoimmune diseases, it mostly involves unknown autoantigens.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Occurrence and Antigenic Specificity of Perinuclear Anti-Neutrophil Cytoplasmic Antibodies (P-ANCA) in Systemic Autoimmune Diseases

Argyropoulou

Goules

Boutzios

et al. 2021

Cells

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“…ANCAs are antibodies directed against proteins located within cytoplasmic granules of neutrophils and monocytes [8,10]. Two key ANCA patterns are described with indirect immunofluorescence (IIF)cytoplasmic staining (C-ANCA), characterised by granular cytoplasmic staining; and peri-nuclear staining (P-ANCA), characterised by homogenous peri-nuclear staining [11]. Although autoantibodies against various different cytoplasmic proteins can produce these patterns, proteinase-3 (PR3) and myeloperoxidase (MPO) have been identified as the clinically relevant autoantigens most strongly associated with systemic vasculitides [8,[11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

“…Two key ANCA patterns are described with indirect immunofluorescence (IIF)cytoplasmic staining (C-ANCA), characterised by granular cytoplasmic staining; and peri-nuclear staining (P-ANCA), characterised by homogenous peri-nuclear staining [11]. Although autoantibodies against various different cytoplasmic proteins can produce these patterns, proteinase-3 (PR3) and myeloperoxidase (MPO) have been identified as the clinically relevant autoantigens most strongly associated with systemic vasculitides [8,[11][12][13][14]. In practice, ANCA are usually detected via initial screening with IIF using ethanol-fixed neutrophils, followed by testing of positive serum via enzymelinked immunosorbent assay (ELISA) to confirm the presence of PR3-ANCA and MPO-ANCA.…”

Section: Introductionmentioning

confidence: 99%

Pulmonary granulomas in a patient with positive ANCA and history of tuberculosis: case report

2020

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Background Granulomatous polyangiitis (GPA) is a rare multisystem autoimmune disease of unknown aetiology that is pathologically characterised by necrotising vasculitis, tissue necrosis and granulomatous inflammation, typically in the presence of anti-neutrophil cytoplasmic antibodies (ANCA). However infectious diseases may induce high titre ANCA and mimic vasculitis. Tuberculosis may share many clinical features with GPA including fever, arthralgia, granulomatous inflammation and pulmonary lesions and patients. Case presentation A 39 year old patient was admitted with ocular irritation and redness, arthralgia and multiple new pulmonary lesions. The past medical history was significant for two episodes of tuberculosis previously requiring prolonged treatment. ANCA antibodies were positive and CT showed multiple pulmonary lesions including cavitatory lesions. After extensive investigation, the patient was treated for GPA with high dose immune suppression with good clinical response. Conclusions Here we review the diagnostic considerations between differentiating GPA and tuberculosis in patients from endemic regions. It is recommended that biopsies of lung lesions, sputum microscopy and multidisciplinary team input are sought as part of the workup when these two differentials are being considered.

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“…Use of both ethanol-fixed neutrophils and formalin-fixed neutrophils improves interpretation of ANCA patterns. 21 In many cases, the atypical patterns are absent or decreased in formalinfixed neutrophils. However, some difficulties arise in testing formalin-fixed neutrophils, including autofluorescence of cells.…”

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confidence: 99%

Variability in Testing for Antineutrophil Cytoplasmic Antibodies: A Survey of Participants in the College of American Pathologists Proficiency Testing Program

Karlon

Naides

Crosson

et al. 2016

Archives of Pathology &Amp; Laboratory Medicine

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Context.-Variability in testing for antineutrophil cytoplasmic antibodies (ANCAs) contributes to confusion and controversy related to testing for vasculitis and other ANCA-associated diseases.Objectives.-To survey laboratory testing practices regarding ANCA testing and to investigate differences in testing algorithms.Design.-Supplemental questions were sent to the 333 laboratories participating in the College of American Pathologists proficiency testing program for ANCA as part of the Special Immunology S2 Survey.Results.-A total of 315 laboratories submitted responses to the supplemental questions. Only 88 of 315 participants (28%) reported using a combination of indirect immunofluorescence (IFA) and enzyme immunoassay (EIA) techniques as recommended by current guidelines, with a few additional labs using IFA and multiplex bead assay as an acceptable alternative to EIA. Other labs reported using only IFA, EIA, or multiplex bead assays.Conclusions.-A wide variety of testing algorithms are in use for ANCA testing despite evidence to suggest that a combination of IFA and EIA testing provides the most comprehensive information. Laboratories should inform clinicians clearly about testing practices and utility of testing in specific disease states.

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A Dual-Fixed Neutrophil Substrate Improves Interpretation of Antineutrophil Cytoplasmic Antibodies by Indirect Immunofluorescence

Abstract: Concomitant staining on formalin fixation of IIF-positive ethanol-fixed ANCA samples improves the interpretation of ANCA testing and is predictive of vasculitis autoantigens MPO and PR3.

Cited by 12 publications

References 19 publications

Occurrence and Antigenic Specificity of Perinuclear Anti-Neutrophil Cytoplasmic Antibodies (P-ANCA) in Systemic Autoimmune Diseases

Occurrence and Antigenic Specificity of Perinuclear Anti-Neutrophil Cytoplasmic Antibodies (P-ANCA) in Systemic Autoimmune Diseases

Pulmonary granulomas in a patient with positive ANCA and history of tuberculosis: case report

Variability in Testing for Antineutrophil Cytoplasmic Antibodies: A Survey of Participants in the College of American Pathologists Proficiency Testing Program

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