Pemphigus Vulgaris IgG Directly Inhibit Desmoglein 3-Mediated Transinteraction

Heupel, Wolfgang-Moritz; Zillikens, Detlef; Drenckhahn, Detlev; Waschke, Jens

doi:10.4049/jimmunol.181.3.1825

Cited by 124 publications

(152 citation statements)

References 48 publications

(71 reference statements)

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“…1 indicates that Dsg3 endocytosis does not require p38 activation and suggests that p38 MAPK activation is secondary to the loss of intercellular adhesion. Taken together, these findings suggest that PV mAbs directly inhibit Dsg3-mediated adhesion, a model that has previously been supported by in vitro assays (14).…”

Section: Discussionsupporting

confidence: 79%

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p38 MAPK Activation Is Downstream of the Loss of Intercellular Adhesion in Pemphigus Vulgaris

Mao

Sano

Park

et al. 2011

Journal of Biological Chemistry

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Pemphigus vulgaris (PV) is a potentially fatal blistering disease characterized by autoantibodies against the desmosomal adhesion protein desmoglein (Dsg) 3. Whether autoantibody steric hindrance or signaling through pathways such as p38 MAPK is primary in disease pathogenesis is controversial. PV mAbs that cause endocytosis of Dsg3 but do not dissociate keratinocytes because of compensatory adhesion by Dsg1 do not activate p38. The same mAbs plus exfoliative toxin to inactivate Dsg1 but not exfoliative toxin alone activate p38, suggesting that p38 activation is secondary to loss of adhesion. Mice with epidermal p38␣ deficiency blister after passive transfer of PV mAbs; however, acantholytic cells retain cell surface Dsg3 compared with wild-type mice. In cultured keratinocytes, p38 knockdown prevents loss of desmosomal Dsg3 by PV mAbs, and exogenous p38 activation causes internalization of Dsg3, desmocollin 3, and desmoplakin. p38␣ MAPK is therefore not required for the loss of intercellular adhesion in PV, but may function downstream to augment blistering via Dsg3 endocytosis. Treatments aimed at increasing keratinocyte adhesion could be used in conjunction with immunosuppressive agents, potentially leading to safer and more effective combination therapy regimens. Pemphigus vulgaris (PV)2 is a potentially fatal autoimmune blistering disorder caused by autoantibodies to keratinocyte cell adhesion proteins known as desmogleins (1). The pathognomonic histologic finding in PV is suprabasal acantholysis, or the detachment of intact keratinocytes from each other because of loss of intercellular adhesion. A characteristic clinical finding in PV is Nikolsky's sign, in which blisters can be induced in otherwise normal-appearing skin by applying pressure or mechanical shear force, reflecting the loss of intercellular adhesion even in skin without overt blisters (2).PV autoantibodies primarily target desmoglein (Dsg) 3, a transmembrane adhesion molecule of desmosomes (3). Passive transfer experiments using neonatal mice have established the pathogenicity of the anti-Dsg3 autoantibodies in PV (4, 5). The anatomic site of blister formation is thought to be due to the tissue-specific expression patterns of different Dsg isoforms, also known as the desmoglein compensation theory. Dsg3 is expressed by basal keratinocytes of mucosa and skin, whereas Dsg1 is expressed by basal keratinocytes in skin but not mucosa (6, 7). Therefore, patients with Dsg3 autoantibodies demonstrate blistering in the mucosa, where compensatory adhesion by Dsg1 is not present (mucosal-dominant PV). In some patients who progress to develop Dsg1 in addition to Dsg3 autoantibodies, suprabasal blisters appear in both the mucosa and skin (mucocutaneous PV) (8 -10).Epitope mapping studies have shown that pathogenic PV autoantibodies preferentially bind the amino-terminal domain of Dsg3 that is predicted to form the transadhesive interface between cells, based on analogy to ultrastructural models for the classical cadherins (11-13). PV IgG has also been shown...

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Section: Discussionsupporting

confidence: 79%

“…PV IgG has also been shown to directly inhibit Dsg3 homophilic interactions in vitro (14). These data have supported the "steric hindrance" model of disease, in which direct interference with Dsg3 interactions causes the loss of intercellular adhesion.…”

Section: Pemphigus Vulgaris (Pv)mentioning

confidence: 59%

p38 MAPK Activation Is Downstream of the Loss of Intercellular Adhesion in Pemphigus Vulgaris

Mao

Sano

Park

et al. 2011

Journal of Biological Chemistry

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“…Cell Culture and Test Reagents-The immortalized human keratinocyte cell line HaCaT was cultivated as described (3). PV antibody AK 23 was purchased from Biozol (Eching, Germany) and used at 75 g/ml for experiments.…”

Section: Methodsmentioning

confidence: 99%

“…Purification and Preparation of Pemphigus IgG and Recombinant Dsg-Fc-Purification of the IgG fractions of a patient with a mucocutaneous PV (PV-IgG 1, both Dsg1 and Dsg3 antibodies) and one with mucous PV (PV-IgG 2, only Dsg3 antibodies), was performed as described previously (3). Final IgG concentrations were adjusted to 150 -500 g/ml for experiments.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, we provided direct evidence that PV autoantibodies directly block Dsg3 transinteraction (3). In contrast, Dsg1 autoantibodies in pemphigus foliaceus (PF) were found to disrupt Dsg1 transinteraction most likely via cellular signaling events rather than by direct inhibition (3,4). However, it still remained unanswered whether direct inhibition of Dsg3 transinteraction is the sole pathogenic mechanism in mucosaldominant PV where autoantibodies against Dsg3 but not against Dsg1 are present (1).…”

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confidence: 99%

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Peptides Targeting the Desmoglein 3 Adhesive Interface Prevent Autoantibody-induced Acantholysis in Pemphigus

Heupel

Müller

Efthymiadis

et al. 2009

Journal of Biological Chemistry

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Pemphigus vulgaris (PV) autoantibodies directly inhibit desmoglein (Dsg) 3-mediated transinteraction. Because cellular signaling also seems to be required for PV pathogenesis, it is important to characterize the role of direct inhibition in pemphigus acantholysis to allow establishment of new therapeutic approaches. Therefore, we modeled the Dsg1 and Dsg3 sequences into resolved cadherin structures and predicted peptides targeting the adhesive interface of both Dsg3 and Dsg1. In atomic force microscopy single molecule experiments, the selfdesigned cyclic single peptide specifically blocked homophilic Dsg3 and Dsg1 transinteraction, whereas a tandem peptide (TP) consisting of two combined single peptides did not. TP did not directly block binding of pemphigus IgG to their target Dsg antigens but prevented PV-IgG-induced inhibition of Dsg3 transinteraction in cell-free (atomic force microscopy) and cell-based (laser tweezer) experiments, indicating stabilization of Dsg3 bonds. Similarly, PV-IgG-mediated acantholysis and disruption of Dsg3 localization in HaCaT keratinocytes was partially blocked by TP. This is the first evidence that direct inhibition of Dsg3 binding is important for PV pathogenesis and that peptidomimetics stabilizing Dsg transinteraction may provide a novel approach for PV treatment. Autoantibodies in pemphigus vulgaris (PV)4 are mainly directed against the adhesion molecules desmoglein (Dsg) 3 and Dsg1 (1, 2). Recently, we provided direct evidence that PV autoantibodies directly block Dsg3 transinteraction (3). In contrast, Dsg1 autoantibodies in pemphigus foliaceus (PF) were found to disrupt Dsg1 transinteraction most likely via cellular signaling events rather than by direct inhibition (3, 4). However, it still remained unanswered whether direct inhibition of Dsg3 transinteraction is the sole pathogenic mechanism in mucosaldominant PV where autoantibodies against Dsg3 but not against Dsg1 are present (1). Alternatively, direct inhibition could act in concert with autoantibody-induced cellular signaling or may just be an epiphenomenon secondary to skin blistering (1, 5-7).The idea of direct interference with Dsg transinteraction by pemphigus autoantibodies has been proposed when Dsg3 was discovered to be a cadherin-type cell adhesion molecule (8, 9). From structural and mutational analyses, it was concluded that classical cadherins are able to form adhesive dimers through their N-terminal cadherin extracellular domain (EC1) via different interaction schemes (10 -13). Investigations of desmosomes using electron tomography from tissue sections also indicated various interaction models for desmosomal cadherins (14). Some of these interaction models resembled the interaction schemes found in the high resolution crystal structure analyses of N-and E-cadherin.This study was designed to modulate Dsg3 and Dsg1 transinteraction by using peptides fitting the most likely Dsg adhesive interface revealed by three-dimensional modeling. In an earlier study, it was reported that peptides targeting the putativ...

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Standardized Production of Anti‐Desmoglein 3 Antibody AK23 for Translational Pemphigus Vulgaris Research

Mueller,

Rahimi,

Sauta

et al. 2024

Current Protocols

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Antibody‐mediated receptor activation is successfully used to develop medical treatments. If the activation induces a pathological response, such antibodies are also excellent tools for defining molecular mechanisms of target receptor malfunction and designing rescue therapies. Prominent examples are naturally occurring autoantibodies inducing the severe blistering disease pemphigus vulgaris (PV). In the great majority of patients, the antibodies bind to the adhesion receptor desmoglein 3 (Dsg3) and interfere with cell signaling to provoke severe blistering in the mucous membranes and/or skin. The identification of a comprehensive causative signaling network downstream of antibody‐targeted Dsg3 receptors (e.g., shown by pharmacological activators or inhibitors) is currently being discussed as a basis to develop urgently needed first‐line treatments for PV patients. Although polyclonal PV IgG antibodies have been used as proof of principle for pathological signal activation, monospecific anti‐Dsg3 antibodies are necessary and have been developed to identify pathological Dsg3 receptor–mediated signal transduction. The experimental monospecific PV antibody AK23, produced from hybridoma cells, was extensively tested in our laboratory in both in vitro and in vivo models for PV and proved to recapitulate the clinicopathological features of PV when generated using the standardized production and purification protocols described herein. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC.Basic Protocol 1: Bovine IgG stripping from FBS and quality controlBasic Protocol 2: AK23 hybridoma expansion and IgG productionBasic Protocol 3: AK23 IgG purificationBasic Protocol 4: AK23 IgG quality controlSupport Protocol 1: Detection of endotoxin levelsSupport Protocol 2: Detection and removal of mycoplasma

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Pemphigus Vulgaris IgG Directly Inhibit Desmoglein 3-Mediated Transinteraction

Cited by 124 publications

References 48 publications

p38 MAPK Activation Is Downstream of the Loss of Intercellular Adhesion in Pemphigus Vulgaris

p38 MAPK Activation Is Downstream of the Loss of Intercellular Adhesion in Pemphigus Vulgaris

Peptides Targeting the Desmoglein 3 Adhesive Interface Prevent Autoantibody-induced Acantholysis in Pemphigus

Standardized Production of Anti‐Desmoglein 3 Antibody AK23 for Translational Pemphigus Vulgaris Research

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