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

Abstract

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

Saliva and Oral Diseases

“…The study of De et al perfectly reproduced this finding and the authors perfectly agreed with the explanation concerning higher Dsg1 levels in severe disease [106]. In contrast to the previously discussed research, one Italian study was designed to assess the use of a BIOCHIP approach compared with ELISA in PV [107]. In fact, the authors considered saliva an unsuitable substrate for autoantibody detection because of the discordance between techniques found when using saliva samples.…”

Comparative Analysis of BIOCHIP Mosaic-based Indirect Immunofluorescence with Direct Immunofluorescence in Diagnosis of Autoimmune Bullous Diseases

“…in their study used the serum and salivary samples to detect anti-Dsg autoantibodies for diagnosis of pemphigus and concluded that saliva is not a suitable sample for BIOCHIP. [ 8 ] Gornowicz-Porowska et al . from Poland compared the original BIOCHIP method with modified BIOCHIP method using monoclonal IgG instead of routine IgG and concluded that modified BIOCHIP has as higher sensitivity and specificity.…”

“…Similar to the previous studies, the current study also revealed that Dsg 3 and BP 180 were the commonly detected antigens using BIOCHIP in pemphigus and pemphigoid groups, respectively [ Table 3 ]. [ 1 4 5 6 8 9 10 11 12 13 ]…”

miRNA Expression Profile of Saliva in Subjects of Yang Deficiency Constitution and Yin Deficiency Constitution

“…Compared with other samples, collection of saliva is convenient, non-invasive, economical and not influenced by the menstrual cycle. For this reason, some studies have focused on using saliva to detect diseases and have showed some clinical benefits [10,11]. Similarly, saliva could be used to improve the comprehensiveness of studies on TCM constitution.…”