Characterization of bullous pemphigoid antigen: A unique basement membrane protein of stratified squamous epithelia

Stanley, John R.; Hawley‐Nelson, Pamela; Yuspa, Stuart H.; Shevach, Ethan M.; Katz, Stephen I.

doi:10.1016/0092-8674(81)90115-x

Cited by 615 publications

(269 citation statements)

References 26 publications

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“…After adding BSA to a final concent.ration of 0.1%, these extracts were preabsorbed with normal human and mouse sera and protein G sepharose (Sigma Chemical Co., St. Louis, MO). Immunoprecipitation was performed with 10/xl of serum or 10 #g of mAb incubated overnight at 4°C, then 25 #1 of protein G incubated for 1 h. Immunoprecipitates were washed 5 times with 0.5 % Triton X-100 in TBS, and precipitated complexes were eluted in SDS gel sample buffer and electrophoresed as previously described (32). In some experiments, the labeled cells were extracted with 1% SDS, 8 M urea in TBS.…”

Section: Immunoblotting and Immunoprecipitationmentioning

confidence: 99%

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Pemphigus vulgaris antigen, a desmoglein type of cadherin, is localized within keratinocyte desmosomes

Kárpáti

Amagai

Prussick

et al. 1993

The Journal of Cell Biology

109

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Abstract. Pemphigus vulgaris antigen (PVA) is a member of the desmoglein subfamily of cadherin cell adhesion molecules. Because autoantibodies in this disease cause blisters due to loss of epidermal cell adhesion, and because desmoglein is found in the desmosome cell adhesion junction, we wanted to determine if PVA is also found in desmosomes. By immunofluorescence, PV IgG bound, in a dotted pattern, to the cell surface of cultured human keratinocytes induced to differentiate with calcium, suggesting junctional staining. However, by preembedding, immunogold electron microscopic studies only slight labeling could be detected in desmosomes, presumably because of difficulty in gold penetration of intact desmosomes. We therefore treated the keratinocytes with 0.01% trypsin in 1 mM calcium, conditions known to preserve cadherin antigenicity but that caused slight separation of desmosomes, before immunogold staining. In this case there was extensive labeling of the extracellular part of desmosomes but not of the interdesmosomal cell membrane which was stained with anti-/32-microglobulin antibodies. To confirm the specificity of this binding we showed that antibodies raised in rabbits against the extracellular portions of PVA also bound desmosomes in these cultures. In intact mouse epidermis we could also show slight, but specific, immunogold desmosomal labeling with PV IgG. Furthermore, neonatal mice injected with PV IgG affinity purified on PVA showed desmosomal separation with the IgG localized to desmosomal cores. These results indicate that PVA is organized and concentrated within the desmosome where it presumably functions to maintain the integrity of stratifying epithelia. p EMPHIGUS vulgaris (PV) ~ is an autoimmune diseasein which blisters of skin and mucous membranes result from loss of epithelial cell-to-cell adhesion, a pathologic process called acantholysis (31). Immunofluorescence studies demonstrate that PV patients have IgG bound on their epidermal cell surfaces in vivo and also have circulating IgG that binds to the cell surface of normal stratified squamous epithelia (5, 6). These PV autoantibodies have been shown to be pathogenic by both clinical and experimental observations (for review see reference 29).The antigen (PVA) recognized by PV autoantibodies has been characterized by immunochemical methods. Immunoprecipitation of cultured keratinocyte and normal human epidermal extracts demonstrates that PV sera bind a 130-kD glycoprotein which forms a stoichiometric complex with plakoglobin (22), a plaque protein of intercellular adhering junctions (i.e., desmosomes and adherens junctions) (8).Recent eDNA cloning has demonstrated that the PVA is a member of the cadherin supergene family (3). The origi-

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Section: Immunoblotting and Immunoprecipitationmentioning

confidence: 99%

“…In some experiments, the labeled cells were extracted with 1% SDS, 8 M urea in TBS. These extracts were dialyzed against 0.3% Nonidet P-40, 2 mM CaC12 in TBS to remove the denaturants, then immunoprecipitation was performed with protein A as previously described (32).…”

Section: Immunoblotting and Immunoprecipitationmentioning

confidence: 99%

Pemphigus vulgaris antigen, a desmoglein type of cadherin, is localized within keratinocyte desmosomes

Kárpáti

Amagai

Prussick

et al. 1993

The Journal of Cell Biology

109

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“…Proteins that might be involved in this linkage are BP230 (Stanley et al, 1981), plectin (Wiche et al, 1984), (Skalli et al, 1994), HD1 (Hieda et al, 1992), and the 200-kDa 6A5 antigen . Plectin and IFAP-300 bind IF in vitro (Foisner et al, 1988;Skalli et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

Integrin alpha 6 beta 4 forms a complex with the cytoskeletal protein HD1 and induces its redistribution in transfected COS-7 cells.

Niessen

Hulsman

Rots

et al. 1997

MBoC

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The integrin a6134 is a major component of hemidesmosomes, in which it is linked to intermediate filaments. Its Subsequent immunoblot analysis revealed that the 500-kDa protein is in fact HD1. In COS-7 cells, which do not express a6f34 or the hemidesmosomal components BP230 and BP180, HD1 is associated with the cytoskeleton, but after transfecting the cells with cDNAs for human a6 and 134, it was, instead, colocalized with a6,64 at the basal side of the cells. The organization of the vimentin, keratin, actin, and tubulin cytoskeletal networks was not affected by the expression of a6f34 in COS-7 cells. The localization of HD1 at the basal side of the cells depends on the same region of 134 that forms a complex containing HD1 in vitro, since the expression of a6 with a mutant 134 subunit that lacks the four fibronectin type III repeats and the connecting segment did not alter the distribution of HD1. The results indicate that for association of a6134 with HD1, the cytoplasmic domain of 134 is essential. We suggest that this association may be crucial for hemidesmosome assembly.

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“…Clinical features of BP include generalized, tense blisters on normal and erythematous skin and, in some cases, blisters and erosions of the mucous membranes (1,2). A hallmark of BP is the presence of circulating autoantibodies against BP180 and BP230, two components of junctional adhesion complexes called hemidesmosomes, critically involved in the maintenance of dermoepidermal adhesion (1,(3)(4)(5)(6)(7)(8)(9)(10).…”

mentioning

confidence: 99%

“…B ullous pemphigoid (BP) 3 is the most frequent autoimmune subepidermal blistering disease of the skin. Clinical features of BP include generalized, tense blisters on normal and erythematous skin and, in some cases, blisters and erosions of the mucous membranes (1,2).…”

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confidence: 99%

Autoreactive T and B Cells from Bullous Pemphigoid (BP) Patients Recognize Epitopes Clustered in Distinct Regions of BP180 and BP230

Thoma-Uszynski

Uter

Schwietzke

et al. 2006

The Journal of Immunology

109

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Bullous pemphigoid (BP) is a well-characterized model of autoantibody-mediated autoimmunity, which presumably depends on autoreactive Th cells that promote the activation of autoreactive B cells. The two major autoantigens of BP are BP180 and BP230, two components of dermoepidermal adhesion complexes. Both, autoreactive Th cell responses and autoantibody profiles were characterized in 35 patients with acute onset BP using BP180 and BP230 proteins. Our findings indicate the following: 1) autoreactive Th cells recognized epitopes within the NH2-terminal (77.1%), COOH-terminal (65.7%), and central portion (57.1%) of the BP180 ectodomain; 2) IgG autoantibodies were found to exhibit similar or identical reactivity against the NH2-terminal (82.8%), COOH-terminal (77.1%), and central portion (37.1%) of the BP180 ectodomain; 3) T and B cell reactivity with the NH2-terminal portion of the BP180 ectodomain was associated with extensive BP, whereas the central portion was more frequently recognized in limited BP; 4) only 7 of 16 (43.7%) and 6 of 16 (37.5%) BP patients showed a Th cellular response against the COOH- and NH2-terminal regions of BP230, respectively, whereas 5) IgG reactivity against the COOH- and NH2-termini of BP230 was detected in 5 of 16 (31.3%) and 6 of 16 (37.5%) patients, respectively. These results demonstrate that Th and B cell reactivities against BP180, are, in contrast to BP230 reactivity, almost constantly detectable in BP patients, and differential epitope recognition of BP180 seems to be associated with distinct clinical severity. These observations support the concept that BP180, but not BP230, is the primary autoantigen of BP critical for disease development.

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Characterization of bullous pemphigoid antigen: A unique basement membrane protein of stratified squamous epithelia

Cited by 615 publications

References 26 publications

Pemphigus vulgaris antigen, a desmoglein type of cadherin, is localized within keratinocyte desmosomes

Pemphigus vulgaris antigen, a desmoglein type of cadherin, is localized within keratinocyte desmosomes

Integrin alpha 6 beta 4 forms a complex with the cytoskeletal protein HD1 and induces its redistribution in transfected COS-7 cells.

Autoreactive T and B Cells from Bullous Pemphigoid (BP) Patients Recognize Epitopes Clustered in Distinct Regions of BP180 and BP230

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