The use of biochip immunofluorescence microscopy for the diagnosis of Pemphigus vulgaris

Russo, Irene; Saponeri, Andrea; Peserico, Andrea; Alaibac, Mauro

doi:10.1016/j.acthis.2013.12.012

Cited by 29 publications

(18 citation statements)

References 13 publications

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“…This is because it is believed that there may be a small risk of prilocaine fetal methemoglobinemia and uterine blood flow disruption that could happen secondary to local anesthetics use during the skin biopsy (Hrgovic, 1990, Nau, 1985, Voorbrood et al, 1982). Meanwhile, newer and safer diagnostic methods, including enzyme-linked immunosorbent assay (ELISA) of BP180 and desmoglein 1 and 3 autoantibodies, indirect immunofluorescence, and biochip immunofluorescence microscopy, should be considered as potential alternatives to biopsying (Russo et al, 2014, Sardy et al, 2013, Tampoia et al, 2012). …”

Section: Management Issuesmentioning

confidence: 99%

Autoimmune blistering diseases in females: a review

Zhao

Murrell

2015

International Journal of Women's Dermatology

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The autoimmune blistering diseases (AIBDs) are a group of heterogeneous skin diseases with autoantibodies directed against structural proteins in the skin. A new interest in the female bias towards autoimmune diseases in general has led to our attention to focus on how and why this female bias manifests in AIBD. The authors aim to review and explore the various aspects of AIBD affecting females more than males, including the higher prevalence, worse quality of life, and complex management issues such as pregnancy and lactation.

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Section: Management Issuesmentioning

confidence: 99%

Autoimmune blistering diseases in females: a review

Zhao

Murrell

2015

International Journal of Women's Dermatology

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“…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).…”

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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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“…ALBIA has several advantages, including requirement of small amount of serum, high‐throughput, short turnaround time and high sensitivity. In addition, a multiplex indirect IF BIOCHIP for pemphigus diseases has recently been developed for Dsg1, Dsg3, BP180 and BP230 .…”

Section: Discussionmentioning

confidence: 99%

Anti‐early endosome antigen 1 autoantibodies were detected in a pemphigus‐like patient but not in the majority of pemphigus diseases

Nishikawa

Takahashi

Matsuda

et al. 2016

Experimental Dermatology

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Although the major autoantigens in classic pemphigus are desmogleins, sera from various types of pemphigus react with a number of other molecules, including desmocollins and plakin proteins. However, other novel pemphigus-related autoantigens remain to be identified. In this study, immunoblotting for serum from an atypical autoimmune bullous disease patient identified an unknown 175 kDa protein. Subsequent studies using two-dimensional gel electrophoresis, immunoblotting and mass-spectrometry identified the 175 kDa protein as early endosome antigen 1 (EEA1). This finding was confirmed by subsequent immunological studies, including indirect immunofluorescence of skin and cultured keratinocytes, two-dimensional gel electrophoresis and immunoblotting with anti-EEA1 polyclonal antibody, and preabsorption with EEA1 recombinant protein. Finally, we developed a novel BIOCHIP assay using full-length EEA1 recombinant protein to detect anti-EEA1 antibodies. However, none of 35 sera from various types of pemphigus showed anti-EEA1 antibodies in the BIOCHIP assay, with the exception of the serum from the index case. In addition, various findings in the index case did not suggest pathogenic role of anti-EEA1 autoantibodies. Therefore, although we successfully identified the 175 kDa protein reacted by a serum of an atypical pemphigus-like patient as EEA1, novel BIOCHIP study for other pemphigus sera indicated that EEA1 is not a common and pathogenic autoantigen in pemphigus.

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The use of biochip immunofluorescence microscopy for the diagnosis of Pemphigus vulgaris

Cited by 29 publications

References 13 publications

Autoimmune blistering diseases in females: a review

Autoimmune blistering diseases in females: a review

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

Anti‐early endosome antigen 1 autoantibodies were detected in a pemphigus‐like patient but not in the majority of pemphigus diseases

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