Detection of Laminin 5-Specific Auto-antibodies in Mucous Membrane and Bullous Pemphigoid Sera by ELISA

Abstract

Mucous membrane pemphigoid (MMP) is an autoimmune bullous disease that primarily affects mucous membranes leading to a scarring phenotype. MMP patients produce auto-antibodies (auto-ab) that preferentially recognize two components of the dermoepidermal basement membrane zone (BMZ): bullous pemphigoid (BP)180 and laminin 5 (LN5). Since detection of disease-specific auto-ab may be critical for diagnosis of MMP, we developed an ELISA with affinity-purified native human LN5. A total of 24 MMP, 72 BP, and 51 control sera were analyzed for LN5-specific auto-ab: 18/24 (75.0%) MMP and 29/72 (40.3%) BP sera were LN5 reactive. Sensitivity and specificity of the LN5 ELISA for MMP were 75% and 84.3%, respectively, and 40.3% and 88.2% for BP, respectively. The LN5 ELISA was more sensitive than a dot blot assay with native LN5, which detected LN5-reactive IgG in 14/24 (58.3%) MMP and 16/72 (22.2%) BP sera. In MMP, but not BP, levels of LN5-reactive IgG correlated with disease severity. Furthermore, IgG reactivity to LN5 of the MMP and BP sera was not significantly associated with IgG reactivity against other autoantigens of the BMZ, such as BP180 or BP230. Thus, the established LN5 ELISA holds great promise as a novel diagnostic and prognostic parameter for MMP.

Prevalence and Clinical Significance of Anti–Laminin 332 Autoantibodies Detected by a Novel Enzyme-Linked Immunosorbent Assay in Mucous Membrane Pemphigoid

“…25 Traditional immunoprecipitation and immunoblot studies were initially used to detect IgG antiLam332 autoantibodies in patients with MMP. 12,14,21,26,27 More recently, enzyme-linked immunosorbent assays (ELISAs) have been developed for such purposes, and early studies 22,23 showed a variable but rather low specificity, since up to 40% of patients with BP may have IgG that is reactive with Lam332. In fact, the prevalence of anti-Lam332 among patients with MMP varied from 31% to 70% in 2 studies.…”

“…In fact, the prevalence of anti-Lam332 among patients with MMP varied from 31% to 70% in 2 studies. 22,28 Using a novel ELISA assay, we analyzed serum samples from 154 patients with MMP and 89 individuals serving as controls. We correlated the results of Lam332 ELISA with immunologic and clinical features to establish the prevalence and clinical significance of Lam332 autoantibody detection in MMP.…”

Brunsting-Perry Pemphigoid of the Scalp with Antibodies against Laminin 332

“…ELISA is a sensitive and specific tool for the detection of anti-laminin-332 antibodies. In the study of Bekou et al [13] , ELISA with native laminin 332 in MMP had a sensitivity of 75% and a specificity of 84.3%. Bekou et al [13] also detected anti-laminin-332 antibodies in 40% of patients with BP.…”

“…In the study of Bekou et al [13] , ELISA with native laminin 332 in MMP had a sensitivity of 75% and a specificity of 84.3%. Bekou et al [13] also detected anti-laminin-332 antibodies in 40% of patients with BP. Anti-laminin-332 antibodies can occur in BP and CP due to the epitope spreading phenomenon; laminin 332 is a ligand for BP180 [14] , and the BP180 COOH terminus is considered as a target antigen in CP.…”

Autoimmune bullous disorders

“…Therefore, dermal protein extracts for immunoblot analysis are used for proof of autoantibodies in EBA. Furthermore, specialised laboratories, including the Departments of Dermatology in Marburg, Freiburg, and Lü beck, carry out additional serological tests in the form of ELISAs, immunoblots, and immunoprecipitation with recombinant and native varieties of the autoantigens Dsg1/Dsg3, BP180/BP230 and laminin-5 (10)(11)(12).…”