Anti-Skin Specific Autoantibodies Detected by a New Immunofluorescence Multiplex Biochip Method in Patients with Autoimmune Bullous Diseases

Tampoia, Marilina; Zucano, Antonietta; Villalta, Danilo; Antico, Antonio; Bizzaro, Nicola

doi:10.1159/000339776

Cited by 40 publications

(49 citation statements)

References 71 publications

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“…Although our study was based on a relatively small group of patients, our results were comparable with previous investigations [10, 12]. Taken together, these cumulative findings indicate that, in the determination of autoantibodies to BP180, the diagnostic specificity of the BIOCHIP method was almost comparable to the ELISA [10, 12], whereas in the detection of anti-BP230-gC the diagnostic sensibility for BP230 was slightly reduced [10, 12].…”

Section: Discussionsupporting

confidence: 89%

“…The BIOCHIP technique has proved to be a specific and sensitive diagnostic alternative to the ELISA diagnosis of BP [10, 12]. A BIOCHIP mosaic, prepared using different tissue sections, cell substrates or antigenic molecules, requires only a simple antibody incubation to obtain a detailed profile, so it is possible to search simultaneously different antibodies directed against different organs or infectious agents.…”

Section: Discussionmentioning

confidence: 99%

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Biochip Technology for the Serological Diagnosis of Bullous Pemphigoid

et al. 2012

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Bullous pemphigoid is an autoimmune blistering skin disease characterized by the presence of circulating autoantibodies which recognize specific proteins of the epidermis and dermoepidermal junction. Diagnosis is based on clinical criteria and laboratory investigations, notably histology, direct and indirect immunofluorescence, and ELISA. This study describes a new immunofluorescence assay for parallel determination of anti-BP180 and anti-BP230 based on recombinant antigenic substrates. The aim of the study was to detect BP180 and BP230 autoantibodies by BIOCHIP technology using both a specially designed recombinant BP180-NC16A protein and cells expressing the BP230-gc antigen fragment. 18 patients with bullous pemphigoid were included in the study. Autoantibodies to BP180 were detected by the BIOCHIP technique in 83.33% of patients with clinical, serological, and immunohistological confirmed bullous pemphigoid while autoantibodies against BP230-gC were detected only in 39% of patients. The detection of anti-BP180-NC16A and anti-BP230-gC by a new biochip-based immunoassay is a suitable alternative to indirect immunofluorescence and ELISA. This method has the advantage of easily discriminating the different autoantibody specificities. The BIOCHIP method is faster, cheaper, and easy to use when compared with the ELISA approach. For this reason, the new method could be used as an initial screening test to identify patients with bullous pemphigoid, and doubtful results could then be confirmed by ELISA.

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Section: Discussionsupporting

confidence: 89%

Section: Discussionmentioning

confidence: 99%

Biochip Technology for the Serological Diagnosis of Bullous Pemphigoid

et al. 2012

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“…For the detection of autoantibodies by BIOCHIP Technology recombinant DSG1 and DSG3 expressing human cells (HEK293 cells) have been used as substrates (3). They are applied to thin glass slides, mechanically cut into millimeter-sized fragments (BIOCHIPs) and then applied automatically onto microscopes slides (3,6,10). The Titerplane Technique, consisting of two consecutive incubation steps (in the first the substrates are incubated with diluted (1/10) patient serum samples, in the second the attached autoantibodies are stained with fluorescein-labeled anti-human antibodies), was the procedure used.…”

Section: Methodsmentioning

confidence: 99%

“…In recent years, a BIOCHIP-based indirect immunofluorescence technique for the determination of anti-DSG3 and anti-DSG1 autoantibodies has been described (3,4). Today, serum anti-DSG3 and anti-DSG1 ELISA is regularly used for the diagnosis and follow up of pemphigus (5,6). Saliva may be also used for a non-invasive diagnosis of this autoimmune skin condition.…”

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confidence: 99%

Salivary Samples for the Diagnosis of Pemphigus vulgaris Using the BIOCHIP Approach: a Pilot Study

Russo

Saponeri

Michelotto

et al. 2017

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Abstract. Pemphigus vulgaris (PV) isPemphigus vulgaris (PV) is a rare autoimmune intraepithelial blistering skin disease characterized by the presence of circulating autoantibodies directed against desmoglein 3 (DSG3) and desmoglein 1 (DSG1), resulting in loss of the normal epithelial cell-to-cell adhesion, through a process called acantholysis (1). The diagnosis of PV is generally based on clinical features, histology and immunological tests, notably direct and indirect immunofluorescence and enzyme linked immunoassorbent assay (ELISA) (2). In recent years, a BIOCHIP-based indirect immunofluorescence technique for the determination of anti-DSG3 and anti-DSG1 autoantibodies has been described (3, 4). Today, serum anti-DSG3 and anti-DSG1 ELISA is regularly used for the diagnosis and follow up of pemphigus (5, 6). Saliva may be also used for a non-invasive diagnosis of this autoimmune skin condition. The use of saliva anti-DSG3 and anti-DSG1 ELISA for the diagnosis of PV has been already reported (7,8). In the present pilot study, we performed both ELISA and BIOCHIP using salivary and serum samples from the same patients to investigate if the detection of anti-desmogleins autoantibodies in salivary samples by BIOCHIP could be used as test for the diagnosis of PV. Patients and MethodsThe study comprised 8 caucasian patients with PV, 3 males and 5 females ( Table I). The diagnosis of PV was based on clinical features, histology and immunopathological findings, notably positive direct immunofluorescence, serum detection of anti-DSG3 and anti-DSG1 autoantibodies by ELISA and serum BIOCHIP. Serum and salivary samples from 2 normal healthy individuals, 1 patient with epidermolysis bullosa acquisita and 2 patients with bullous pemphigoid were used for negative controls (Table II).For the detection of autoantibodies, an ELISA assay was employed using recombinant proteins DSG1 and DSG3 (MBL, Nagoya, Japan), made of the entire extracellular domain of DSG1 and DSG3 respectively and produced by Amagai et al. (9) with cut off values of 20 U/ml for DSG1 and DSG3. For the detection of autoantibodies by BIOCHIP Technology recombinant DSG1 and DSG3 expressing human cells (HEK293 cells) have been used as substrates (3). They are applied to thin glass slides, mechanically cut into millimeter-sized fragments (BIOCHIPs) and then applied 97

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“…In autoimmune blistering diseases, a BIOCHIP ® mosaic containing six substrates comprising monkey esophagus, salt-split skin, recombinant BP180 NC16A, and HEK293 cells transfected for expression of desmoglein 1, desmoglein 3, and BP230 is widely used in the dermatological community [136,[153][154][155][156].…”

Section: Biochip ® Mosaicmentioning

confidence: 99%

Clinical features and diagnosis of epidermolysis bullosa acquisita

Vorobyev

Ludwig

Schmidt

2016

Expert Review of Clinical Immunology

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Introduction: Epidermolysis bullosa acquisita (EBA) is a rare autoimmune blistering disease of skin and mucous membranes. EBA is caused by autoantibodies against type VII collagen, which is a major component of anchoring fibrils, attaching epidermis to dermis. Binding of autoantibodies to type VII collagen leads to skin fragility and, finally, blister formation. The clinical picture of EBA is polymorphic, with several distinct phenotypes being described. Despite recent progress in understanding the pathophysiology of EBA, its diagnosis is still challenging. Areas covered: This review provides an update on the clinical manifestations and diagnostic methods of EBA. We searched PubMed using the terms 'epidermolysis bullosa acquisita' covering articles in English between 1 January 2005 and 31 May 2016. Relevant older publications were retrieved form cited literature. Expert commentary: While the clinical picture is highly variable, diagnosis relies on direct immunofluorescence (IF) microscopy of a perilesional skin biopsy. Linear deposits of IgG, IgA and/or C3 along the dermal-epidermal junction with an u-serrated pattern are diagnostic for EBA alike the detection of serum autoantibodies against type VII collagen. Several test systems for the serological diagnosis of EBA have recently become widely available. In some patients, sophisticated diagnostic approaches only available in specialized centers are required. ARTICLE HISTORY

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Anti-Skin Specific Autoantibodies Detected by a New Immunofluorescence Multiplex Biochip Method in Patients with Autoimmune Bullous Diseases

Cited by 40 publications

References 71 publications

Biochip Technology for the Serological Diagnosis of Bullous Pemphigoid

Biochip Technology for the Serological Diagnosis of Bullous Pemphigoid

Salivary Samples for the Diagnosis of Pemphigus vulgaris Using the BIOCHIP Approach: a Pilot Study

Clinical features and diagnosis of epidermolysis bullosa acquisita

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