Journal: Current Molecular Medicine

Article Title: Molecular Diagnosis in Autoimmune Skin Blistering Conditions

doi: 10.2174/15665240113136660079

Figure Lengend Snippet: Quantitative Immunoassays for the Detection of Autoantibodies in Autoimmune Blistering Skin Diseases

Article Snippet: ELISA systems using recombinant BP180 and collagen VII have been developed for measuring IgG and IgA autoantibodies (Fig. ) [ ].

Techniques:

Journal: Current Molecular Medicine

Article Title: Molecular Diagnosis in Autoimmune Skin Blistering Conditions

doi: 10.2174/15665240113136660079

Figure Lengend Snippet: Characteristic findings in paraneoplastic pemphigus. (a) Hemorrhagic erosions with crusts on the lips and oral cavity of a patient with non-Hodgkin lymphoma. (b) Interface dermatitis by histopathology (H&E staining). (c) Immunofluorescence (IF) microscopy analysis of a perilesional skin biopsy reveals deposits of IgG at the dermal-epidermal junction and at the intercellular spaces of keratinocytes. (d) Further serological testing by indirect IF microscopy shows binding of IgG autoantibodies to rat bladder urothelium (magnification, 200x). (e) Extract of cultured keratinocytes fractionated by 7.5% SDS-PAGE was transferred to nitrocellulose, and incubated with sera from patients with pemphigus vulgaris (lane 1), bullous pemphigoid (lane 2), paraneoplastic pemphigus serum (lane 3), and normal human serum (lane 4). PNP serum reacts specifically with envoplakin (210 kD, upper arrow) and periplakin (190 kD, lower arrow). The control serum shows no specific reactivity. Migration position for BP230, BP180, Desmoglein (Dsg) 1 and 3 are depicted on the left side of the panel.

Article Snippet: ELISA systems using recombinant BP180 and collagen VII have been developed for measuring IgG and IgA autoantibodies (Fig. ) [ ].

Techniques: Histopathology, Staining, Immunofluorescence, Microscopy, Binding Assay, Cell Culture, SDS Page, Incubation, Control, Migration

Journal: Current Molecular Medicine

Article Title: Molecular Diagnosis in Autoimmune Skin Blistering Conditions

doi: 10.2174/15665240113136660079

Figure Lengend Snippet: Diagnostic Criteria for Bullous Pemphigoid

Article Snippet: ELISA systems using recombinant BP180 and collagen VII have been developed for measuring IgG and IgA autoantibodies (Fig. ) [ ].

Techniques: Diagnostic Assay, Immunofluorescence, Microscopy, Binding Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Current Molecular Medicine

Article Title: Molecular Diagnosis in Autoimmune Skin Blistering Conditions

doi: 10.2174/15665240113136660079

Figure Lengend Snippet: Molecular specificity of autoantibodies in bullous pemphigoid (BP). (a) Sera from patients with BP were tested by ELISA using a recombinant form of the 16th noncollagenous (NC16) A domain of the bullous pemphigoid (BP) antigen 180 and with recombinant BP230 as described [92, 93]. The dotted line represents the cut-off of the assay. (b) An extract of cultured keratinocytes was fractionated by 7.5% SDS-PAGE, transferred to nitrocellulose, and incubated with a BP180-specific monoclonal antibody (lane 1), serum from patients with BP (lanes 2–3), and serum from a healthy control (NHS; lane 4). The BP serum in lane 2 reacts with bullous pemphigoid antigen 180 (BP180, 180kDa, lower arrow) only. The BP serum depicted in lane 3 reacts with both bullous pemphigoid antigen 230 (BP230, 230 kDa, upper arrow) and BP180. Control serum shows no specific reactivity (lane 4).

Article Snippet: ELISA systems using recombinant BP180 and collagen VII have been developed for measuring IgG and IgA autoantibodies (Fig. ) [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Cell Culture, SDS Page, Incubation, Control

Journal: Current Molecular Medicine

Article Title: Molecular Diagnosis in Autoimmune Skin Blistering Conditions

doi: 10.2174/15665240113136660079

Figure Lengend Snippet: Diagnostic Criteria for Mucous Membrane Pemphigoid

Article Snippet: ELISA systems using recombinant BP180 and collagen VII have been developed for measuring IgG and IgA autoantibodies (Fig. ) [ ].

Techniques: Diagnostic Assay, Membrane, Immunofluorescence, Microscopy, Binding Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Current Molecular Medicine

Article Title: Molecular Diagnosis in Autoimmune Skin Blistering Conditions

doi: 10.2174/15665240113136660079

Figure Lengend Snippet: Diagnostic Criteria for Linear IgA Disease

Article Snippet: ELISA systems using recombinant BP180 and collagen VII have been developed for measuring IgG and IgA autoantibodies (Fig. ) [ ].

Techniques: Diagnostic Assay, Immunofluorescence, Microscopy, Binding Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Current Molecular Medicine

Article Title: Molecular Diagnosis in Autoimmune Skin Blistering Conditions

doi: 10.2174/15665240113136660079

Figure Lengend Snippet: Molecular specificity of IgA autoantibodies in pemphigoid diseases. (a) Spent medium of cultured keratinocytes was concentrated by ammonium sulfate precipitation, electrophoretically separated by 8% SDSPAGE, transferred on nitrocellulose and immunoblotted with serum from linear IgA disease (LAD) patients (lanes 1-3) and control serum (NHS) (lane 4). Bound autoantibodies were visualized using a peroxidase-labeled secondary antibody specific for human IgA. The shed ectodomain of BP180 of 120 kDa (LAD-1) and its 97 kDa degradation product (LABD97) are indicated by arrow and arrow head, respectively. (b) ELISA reactivity with the recombinant BP180 ectodomain of IgA autoantibodies from LAD, bullous pemphigoid (BP), dermatitis herpetiformis (DH) patients and age-matched, healthy donors (NHS). The cut-off of the assay is represented by a dashed line. Scatter plot adapted from [138].

Article Snippet: ELISA systems using recombinant BP180 and collagen VII have been developed for measuring IgG and IgA autoantibodies (Fig. ) [ ].

Techniques: Cell Culture, Control, Labeling, Enzyme-linked Immunosorbent Assay, Recombinant

Journal: Current Molecular Medicine

Article Title: Molecular Diagnosis in Autoimmune Skin Blistering Conditions

doi: 10.2174/15665240113136660079

Figure Lengend Snippet: Major Autoantigens in Bullous Diseases

Article Snippet: ELISA systems using recombinant BP180 and collagen VII have been developed for measuring IgG and IgA autoantibodies (Fig. ) [ ].

Techniques: Immunopeptidomics, Membrane

Journal: JCI Insight

Article Title: Implicating bites from a leishmaniasis sand fly vector in the loss of tolerance in pemphigus

doi: 10.1172/jci.insight.123861

Figure Lengend Snippet: The ELISA plates were coated with the recombinant proteins Dsg1 and Dsg3 or other epidermal proteins; BP180, desmocollin 3 and desmoplakin and then incubated with a decreasing concentration of rPpSP32, the protein of interest or CFP32, a control protein. The plates were then incubated in the presence of human sera with anti-PpSP32 or -CFP32 antibodies. The results are representative of 3 independent experiments. Significant differences were found between the different PpSP32 concentrations for Dsg1 and Dsg3 ( P = 0.006 and P = 0.008 between the condition 2 μg/mL and 0.0002 μg/mL for Dsg1 and Dsg3, respectively). For the control proteins, no significant difference was obtained while decreasing the concentration of PpSP32 ( P > 0.05).

Article Snippet: The 96-well microtiter plates (Nunc, Maxisorp) were coated after optimization with 0.1 μg recombinant human proteins, Dsg1 (MyBioSource MBS2010418), Dsg3 (MyBioSource MBS2010322), BP180 (MyBioSource MBS2012222), desmocollin 3 (MyBioSource MBS637447), desmoplakin (MyBioSource MBS2031176), rPpSP32, or a control protein (recombinant form of the CFP32 protein of Mycobacterium tuberculosis ) ( ) in a carbonate-bicarbonate buffer (0.1 M, pH 9.6) at 4°C overnight.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Incubation, Concentration Assay

Journal: Journal of the American Academy of Dermatology

Article Title: Comparison of C3d immunohistochemical staining to enzyme-linked immunosorbent assay and immunofluorescence for diagnosis of bullous pemphigoid

doi: 10.1016/j.jaad.2020.02.020

Figure Lengend Snippet: BP180 and BP230 levels via enzyme-linked immunosorbent assay testing compared with C3d, direct immunofluorescent ( DIF ), and indirect immunofluorescent ( IIF ) results

Article Snippet: Patients with at least 1 positive BP180 or BP230 ELISA, or both, in the setting of clinical concern for BP (based on clinical documentation) and a skin biopsy specimen within 1 year of ELISA testing, between 2010 and 2014, were identified from our dermatopathology database NewPath.

Techniques: Enzyme-linked Immunosorbent Assay