Ca signalling is critical for autoantibody-induced blistering of human epidermis in pemphigus.

BACKGROUND

Pemphigus is a severe bullous autoimmune skin disease. Pemphigus foliaceus (PF) is characterized by antidesmoglein (Dsg) 1 IgG causing epidermal blistering; mucosal pemphigus vulgaris (mPV) by anti-Dsg3 IgG inducing erosions in the mucosa; and mucocutaneous pemphigus vulgaris (PV) by affecting both, with autoantibodies targeting Dsg1 and Dsg3.

OBJECTIVES

To characterize the Ca flux pathway and delineate its importance in pemphigus pathogenesis and clinical phenotypes caused by different antibody profiles.

METHODS

Immunoprecipitation, Ca flux analysis, Western blotting, immunofluorescence staining, dissociation assays and a human skin ex vivo model were used.

RESULTS

PV IgG and PF IgG, but neither Dsg3-specific monoclonal antibody (AK23) nor mPV IgG, caused Ca influx in primary human keratinocytes. Phosphatidylinositol 4-kinase α interacts with Dsg1 but not with Dsg3. Its downstream target - phospholipase-C-γ1 (PLC) - was activated by PV IgG and PF IgG but not AK23 or mPV IgG. PLC releases inositol 1,4,5-trisphosphate (IP3) causing IP3 receptor (IP3R) activation and Ca flux from the endoplasmic reticulum into the cytosol, which stimulates Ca release-activated channels (CRAC)-mediated Ca influx. Inhibitors against PLC, IP3R and CRAC effectively blocked PV IgG and PF IgG-induced Ca influx; ameliorated alterations of Dsg1 and Dsg3 localization, and reorganization of keratin and actin filaments; and inhibited loss of cell adhesion in vitro. Finally, inhibiting PLC or IP3R was protective against PV IgG-induced blister formation and redistribution of Dsg1 and Dsg3 in human skin ex vivo.

CONCLUSIONS

Ca -mediated signalling is important for epidermal blistering and dependent on the autoantibody profile, which indicates different roles for signalling complexes organized by Dsg1 and Dsg3. Interfering with PLC and Ca signalling may be a promising approach to treat epidermal manifestations of pemphigus.