Abstract
Pemphigus Vulgaris is an autoimmune disease that results in blister formation in the epidermis and in mucosal tissues due to antibodies recognizing desmosomal cadherins, mainly desmoglein-3 and -1. Studies on the molecular mechanisms of Pemphigus have mainly been carried out using the spontaneously immortalized human keratinocyte cell line HaCaT or in primary keratinocytes. However, both cell systems have suboptimal features, with HaCaT cells exhibiting a large number of chromosomal aberrations and mutated p53 tumor suppressor, whereas primary keratinocytes are short-lived, heterogeneous and not susceptible to genetic modifications due to their restricted life-span. We have here tested the suitability of the commercially available human keratinocyte cell line hTert/KER-CT as a model system for research on epidermal cell adhesion and Pemphigus pathomechanisms. We here show that hTert cells exhibit a calcium dependent expression of desmosomal cadherins and are well suitable for typical assays used for studies on Pemphigus, such as sequential detergent extraction and Dispase-based dissociation assay. Treatment with Pemphigus auto-antibodies results in loss of monolayer integrity and altered localization of desmoglein-3, as well as loss of colocalization with flotillin-2. Our findings demonstrate that hTert cells are well suitable for studies on epidermal cell adhesion and Pemphigus pathomechanisms.
Highlights
Pemphigus vulgaris (PV) and Pemphigus foliaceus (PF) are blistering autoimmune diseases of the skin and/or oral mucosa that result from loss of cell-cell adhesion of the epidermal/mucosal cells
We have here studied the suitability of the immortalized human keratinocyte cell line hTert/KER-CT as a model system for studies on desmosomal adhesion and Pemphigus pathogenesis. We show that these cells express desmogleins in a calcium dependent manner, whereas the expression of flotillins, which we have shown to interact with Dsg3 [9], is calcium independent
We here aimed at testing the suitability of the recently generated, commercially available human keratinocyte cell line hTert (KER-CT, ATCC CRL-4048) as a model system to study the mechanisms of loss of desmosomal cell adhesion in Pemphigus
Summary
Pemphigus vulgaris (PV) and Pemphigus foliaceus (PF) are blistering autoimmune diseases of the skin and/or oral mucosa that result from loss of cell-cell adhesion of the epidermal/mucosal cells. In Pemphigus, autoantibodies against desmosomal components, mainly desmoglein-3 (Dsg3) and -1 (Dsg1) cause disruption of desmosomal cell adhesion, resulting in blistering of the epidermis and/or mucosa (for a review, please see [1]). The molecular events in keratinocytes that result in the loss of desmosomal cell adhesion have been actively studied for a number years. It has been shown that pathogenic antibodies against desmogleins induce active intracellular signaling events, which may indirectly result in loss of desmosomal cell adhesion. The molecular mechanisms of desmosomal adhesion and signaling in Pemphigus have recently been reviewed by Spindler and Waschke et al, and we would like to refer the readers to these reviews for a more detailed description [2,3]
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