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PubMed1983

Ultrastructure of uninvolved oral mucosa in pemphigus patients.

The uninvolved oral mucosa of seven pemphigus patients was compared with that of age- and sex-matched controls. Three patients had pemphigus erythematodes, two had pemphigus vulgaris, one had pemphigus foliaceus, and one pemphigus vegetans. Five out of seven pemphigus patients demonstrated wider intercellular spaces than the controls. This difference was seen in both basal and spinous epithelial cell layers and was more pronounced in the basal cell layer. There were fewer desmosomes and microvilli than in the controls. It appears that ultrastructural changes in pemphigus also occur in the uninvolved oral mucosa of pemphigus patients.

Adult↗
PubMed1998

Pemphigus in Pakistan, a study of 108 cases [corrected].

Pemphigus has an average age of onset in the fifth and sixth decades of life, with a mean age of onset of about 50 to 60 years. In this study of 108 patients conducted in Karachi, Pakistan, 82 had pemphigus vulgaris, 24 pemphigus foliaceus and 2 pemphigus vegetans. Other types of pemphigus were not seen. Seventy cases were under 41 years with the mean age at the time of diagnosis being 33.8 years, which is considerably low compared to that mentioned in standard western literature. Considerable variation in the age incidence of the disease exists in different geographic locations.

Adolescent↗
PubMed1993

Autoimmune pemphigus. A distinct staining pattern with an anti-desmoglein antibody.

BACKGROUND AND DESIGN: Diagnosis of autoimmune pemphigus is based on the immunodetection of IgG deposits in the epidermal intercellular spaces. Desmoglein is a desmosomal component that has been assumed to be the antigen of pemphigus foliaceus. We investigated the use of one monoclonal antibody (32-2B) to desmoglein 1 on paraffin sections. Twenty-nine pemphigus samples were studied (22 pemphigus vulgaris, seven pemphigus foliaceus). RESULTS: In 29 patients suffering from autoimmune pemphigus, the staining for desmoglein was represented by coarse granules along the cytoplasmic borders of epithelial cells in lesional and perilesional skin and in mucous membranes. This peculiar staining is different from the fine dots along the cytoplasmic membrane observed in normal skin. This abnormal staining seems to be specific for pemphigus. Hereditary acantholytic diseases and other diseases such as bullous pemphigoid, intraepidermal IgA dermatosis, eczema, herpes, and transient acantholytic dermatosis show a similar pattern to normal skin. CONCLUSION: The use of 32-2B on biopsy specimens in cases of autoimmune pemphigus demonstrated a specific staining pattern. It could be used on paraffin sections as a diagnostic test of autoimmune pemphigus. It can even be done retrospectively.

Antibodies, Monoclonal↗
PubMed1991

Immunoblot assay as an aid to the diagnoses of unclassified cases of pemphigus.

With use of an immunoblot assay for both the normal human epidermal extracts and the bovine desmosome preparation, we investigated sera from 18 patients with pemphigus, who exhibited atypical clinical or histologic features and whose diagnoses were difficult to identify. Four sera yielded a 130-kd protein band in the epidermal extracts, which is characteristic of pemphigus vulgaris. A 150-kd protein was identified by nine sera in the epidermal extracts and/or the desmosome preparation, which is known as a pemphigus foliaceus antigen. Five sera demonstrated neither antigen. These results suggest that the immunoblot technique is useful to distinguish pemphigus vulgaris from pemphigus foliaceus and can be a good tool for the diagnosis of pemphigus, especially unclassified cases.

Humans↗
PubMedDate not supplied

The tryptic cleavage product of the mature form of the bovine desmoglein 1 ectodomain is one of the antigen moieties immunoprecipitated by all sera from symptomatic patients affected by a new variant of endemic pemphigus.

Multiple antigens are recognized by sera from patients with pemphigus foliaceus (PF). Several have been identified including keratin 59, desmocollins, envoplakin, periplakin, and desmogleins 1 and 3 (Dsg1 and Dsg3). In addition, an 80 kDa antigen was identified as the N-terminal fragment of Dsg1 using as antigen source an insoluble epidermal cell envelope preparation. However, still unsolved was the identity of the most important antigenic moiety, a 45 kDa tryptic fragment which is recognized by all sera from patients with fogo selvagem, pemphigus foliaceus, by half of pemphigus vulgaris sera and by a new variant of endemic pemphigus in E1 Bagre, Colombia that resembles Senear-Usher syndrome. Here, we report the identification of the 45 kDa conformational epitope of a soluble tryptic cleavage product from viable bovine epidermis. To elucidate the nature of this peptide, viable bovine epidermis was trypsin-digested, and glycosylated peptides were partially purified on a concanavalin A (Con-A) affinity column. This column fraction was then used as an antigen source for further immunoaffinity purification. A PF patient's serum covalently coupled to a Staphylococcus aureus protein A column was incubated with the Con-A eluted products and the immuno-isolated antigen was separated by SDS-PAGE, transferred to a membrane, and visualized with Coomassie blue, silver and amido black stains. The 45 kD band was subjected to amino acid sequence analysis revealing the sequence, EXIKFAAAXREGED, which matched the mature form of the extracellular domain of bovine Dsg1. This study confirms the biological importance of the ectodomain of Dsg1 as well as the relevance of conformational epitopes in various types of pemphigus.

Animals↗
PubMed2003

Detection of pemphigus autoantibodies by IIF and ELISA tests in patients with pemphigus vulgaris and foliaceus and in healthy relatives.

BACKGROUND: Pemphigus is a life-threatening, autoimmune blistering disease, mediated by IgG autoantibodies. The aim of our study was to assess the usefulness of a new enzyme-linked immunosorbent assay (ELISA) in detecting circulating pemphigus autoantibodies, and to compare its sensitivity and specificity with the indirect immunofluorescence (IIF) test. We also established the frequency of occurrence of pemphigus autoantibodies in relatives of our patients. MATERIAL/METHODS: IIF and ELISA tests were performed in 24 patients with pemphigus vulgaris, 13 with pemphigus foliaceus, 56 healthy relatives, and 50 controls, selected according to sex and age. RESULTS: The obtained results revealed high specificity and sensitivity of ELISA, comparable to the IIF test, especially in patients who were in the active stage of the disease. We also showed that the profile of anti-Dsg 1 and/or anti-Dsg 3 autoantibodies is associated with the clinical variant of pemphigus. The frequency of occurrence of pemphigus autoantibodies in the relatives (24/55) performed by IIF was significantly higher (p<0.001) than in the controls (0/50). The same antibodies detected by ELISA (11/56) were less frequent. CONCLUSIONS: In clinically doubtful cases, in which autoantibodies titres by IIF are equal on both substrates (monkey and guinea pig esophagus), the assessment of the exact antibody profile (anti-Dsg 1 and/or anti-Dsg 3) is important to establish the subtype of pemphigus. The frequency of pemphigus antibody occurrence in healthy relatives seems not to be incidental, but further studies should be performed to explain this phenomenon.

Adolescent↗
PubMed1990

Ultrastructural localization of pemphigus vulgaris antigen on canine keratinocytes in vivo and in vitro.

Pemphigus antigens were localized, by use fo immunoelectron microscopy, on canine keratinocytes in vivo on esophageal mucosa and in vitro on established cultured keratinocytes. Convalescent sera from a human being with pemphigus vulgaris and a human being with pemphigus foliaceus reacted with the interdesmosomal cytoplasmic keratinocyte membrane of canine esophagus. Cultured canine keratinocytes expressed the pemphigus vulgaris antigen in a similar pattern, but did not carry the pemphigus foliaceus antigen. The differential presence of cell surface antigens and its relation to various forms of the disease are discussed.

Animals↗
PubMed1997

Postpemphigus acanthomata: a sign of clinical activity?

BACKGROUND: Pemphigus is a group of vesiculobullous disorders in which the blisters usually heal with hyper or hypopigmentation. The appearance of acanthomata at sites of previous blisters has been noted in some cases. METHODS: All cases of pemphigus admitted to the Madras Medical College hospitals during a 2-year period from March 1993 to March 1995 were taken into the study and screened for the presence of acanthomata. RESULTS: Fifty-two cases of pemphigus were identified, 47 of pemphigus vulgaris and five of pemphigus foliaceus; and of these 13 developed acanthomata when the blisters healed. Ten of these cases were of pemphigus vulgaris and three were of pemphigus foliaceus; biopsy of these lesions showed hyperkeratosis, acanthosis, papillomatosis, and intraepidermal clefting. Immunofluorescence carried out in two of these acanthomata also showed intercellular fluorescence. CONCLUSIONS: The occurrence of acanthomata in healed lesions of pemphigus is not uncommon; because histopathologic and immunofluorescence evidence of disease activity is present, cases of this sort require careful follow-up.

Adult↗
PubMed1995

Demonstration of antibodies to bovine desmocollin isoforms in certain pemphigus sera.

We have shown previously that IgG antibodies in certain pemphigus sera, particularly endemic Brazilian pemphigus foliaceus (BPF) sera, react with bovine desmocollins (Dsc), which are transmembranous glycoproteins of desmosome junctions. Desmocollins occur as three different isoforms (Dsc 1, 2 and 3), all of which are represented in the epidermis. In this study, we examined sera of various pemphigus types by immunoblotting purified bovine desmosomes and bovine Dsc 1, 2 and 3 fusion proteins, expressed in pGEX expression vectors. Six of 15 (40.0%) BPF sera, two of 18 (11.1%) non-endemic pemphigus foliaceus sera, eight of 39 (20.5%) pemphigus vulgaris (PV) sera, and two of 11 (18.2%) normal sera, showed reactivity with Dsc from desmosomes. Experiments with fusion proteins showed that no Dsc isoform was specifically recognized by sera of any individual pemphigus type. Our results indicate that the pathogenesis of pemphigus might be more complex than previously believed.

Animals↗
PubMed1997

T lymphocytes from a subset of patients with pemphigus vulgaris respond to both desmoglein-3 and desmoglein-1.

Pemphigus vulgaris and pemphigus foliaceus are cutaneous autoimmune diseases characterized by intraepithelial blisters and autoantibodies to desmosomal glycoproteins. The antigens recognized by pemphigus vulgaris and pemphigus foliaceus autoantibodies are desmoglein-3 (Dsg3) and desmoglein-1 (Dsg1), respectively. Dsg3 and Dsg1 are members of the desmoglein subfamily of the cadherin supergene family of cell adhesion molecules. It has been well documented that a subset of pemphigus vulgaris sera have IgG reactivity to both Dsg1 and Dsg3, suggesting that Dsg1 may also participate in the autoimmune response of these patients. The cellular mechanisms of T cell autoimmunity in these patients, however, are completely unknown. In this study, we tested the proliferative responses of T lymphocytes from eight pemphigus vulgaris patients after incubation with Dsg3 and Dsg1 fusion proteins. The sera of four of these PV patients showed reactivity with both Dsg1 and Dsg3, whereas the remaining four reacted only with Dsg3. We found that T cells obtained from those patients that exhibited the combined Dsg1/Dsg3 autoantibody reactivity showed a proliferative response after exposure to either Dsg1 or Dsg3 fusion proteins. The cellular responses to both of these recombinant proteins were highly specific and restricted to the CD4-positive T cell population. T cells from pemphigus vulgaris patients with no anti-Dsg1 serum reactivity showed a proliferative response to Dsg3, but not to Dsg1. The Dsg1 fusion protein used in this study has minimal sequence homology with Dsg3. Thus, this study provides the first evidence that T cells from a subset of pemphigus vulgaris patients respond to both Dsg1 and Dsg3.

Amino Acid Sequence↗
PubMed1996

Expression of major histocompatibility complex class II molecules by dermal inflammatory cells, epidermal Langerhans cells and keratinocytes in canine dermatological disease.

The distribution of Major Histocompatibility Complex (MHC) Class II molecules in skin biopsies from 42 dogs with primary skin disease and six control dogs was examined by indirect immunohistochemical methods. Canine epidermal Langerhans cells constitutively expressed MHC Class II and the number of such cells was increased in the epidermis of dogs with discoid lupus erythematosus (n = 10), hypersensitivity dermatitis (n = 10) or deep pyoderma (n = 13) but not in eight cases of pemphigus foliaceus. Expression of MHC Class II by epidermal and follicular keratinocytes was observed in a ninth dog with pemphigus foliaceus. Within dermal inflammatory infiltrates, Class II was expressed by macrophages, dendritic cells and fibroblasts but not by neutrophils, eosinophils, mast cells or plasma cells. Only occasional dermal lymphocytes expressed surface MHC Class II, but nodular aggregates of strongly Class II-positive lymphocytes were observed within the dermis of two dogs with discoid lupus erythematosus.

Animals↗
PubMed1985

The longevity of immunoglobulin preservation in canine skin utilizing Michel's fixative.

Skin biopsy specimens from 7 dogs with immune-mediated skin diseases diagnosed by routine histology and 5 dogs with other skin diseases were placed in Michel's transport medium for 4 to 9 years. Direct immunofluorescence yielded positive results in tissue samples from 3 dogs with pemphigus foliaceus and 2 dogs with discoid lupus erythematosus. Direct immunofluorescence was not seen in tissue samples from 1 dog with pemphigus foliaceus and 5 dogs with non immune-mediated skin diseases. Direct immunofluorescence was seen in skin biopsy specimens maintained in Michel's medium for 4 to 8 years.

Animals↗
PubMed1994

Unusual acantholytic bullous dermatosis associated with neoplasia and IgG and IgA antibodies against bovine desmocollins I and II.

There are unusual cases of pemphigus that have antibodies nonreactive with either pemphigus vulgaris or pemphigus foliaceus antigens. We describe a patient with an acantholytic bullous dermatosis and lung cancer with intercellular IgG and IgA antibodies that differed in specificity from those of pemphigus vulgaris and pemphigus foliaceus and reacted with bovine desmocollins I and II and recombinant desmocollin protein.

Adult↗
PubMed2004

A sensitive and restricted enzyme-linked immunosorbent assay for detecting a heterogeneous antibody population in serum from people suffering from a new variant of endemic pemphigus.

We recently described a new variant of endemic pemphigus foliaceus (EPF) in El Bagre, Colombia, that resembles Senear-Usher syndrome and identified autoantibodies to desmoglein 1 (Dsg1), as well as to multiple known and unknown antigens including plectins, in the serum of these patients. Here, we developed a cost-effective ELISA assay capable of detecting the heterogeneous antibody population observed in these EPF patients, and useful for serum epidemiological studies. A protein extract obtained from trypsin-digested fresh bovine skin and further purified on a concanavalin A matrix was used as antigen. This extract contains an important conformational epitope (a 45 kDa tryptic fragment of the Dsg1 ectodomain), which is recognized by antibodies in serum from patients with all varieties of pemphigus foliaceus (PF), and from half of those with pemphigus vulgaris with active clinical disease. The cut-off and threshold values were normalized using human serum obtained from both endemic and non-endemic areas for PF. The efficiency of this ELISA was tested using 600 serum samples from controls and patients diagnosed with EPF, non-endemic PF and other bullous diseases. The overall sensitivity and specificity of the assay were determined to be 95% and 72%, respectively, with reproducibilities of 98% (intraassay) and 95% (interassay). Comparing the ELISA with other tests to detect EPF autoantibodies, this ELISA was the most sensitive, followed by direct immunofluorescence (DIF), indirect immunofluorescence using anti-IgG4 monoclonal antibodies and immunoprecipitation (IP), respectively. The most specific assay was IP, followed by DIF. Immunoblotting to Dsg1 exhibited both poor sensitivity and poor specificity, although plectins were well visualized. We conclude that this ELISA is an excellent tool for field serological studies, allowing testing of multiple serum samples simultaneously and for detecting, with appropriate restriction and sensitivity, the heterogeneous antibody population seen in patients with this variant of EPF. Finally, autoantibody serum levels obtained with this ELISA correlated well with the clinical activity and extent of disease in patients with El Bagre EPF.

Animals↗
PubMed1976

Pemphigus controlled by dapsone.

Three uncomplicated cases of pemphigus were clinically controlled by dapsone. Improvement was associated with decreasing titres of circulating intercellular antibodies. Interestingly, the sera from one case of pemphigus foliaceus contained intercellular antibodies found in the Malpighian and basal cell layers using the fluorescent technique and in the granular layer using the peroxidase technique. These findings suggest that the intercellular antibodies in pemphigus vulgaris and pemphigus foliaceus are similar but bind at different anatomical sites.

Adult↗
PubMed2005

Two cases of feline leishmaniosis in Switzerland.

Two cats with Leishmania species infections were investigated. The first had been imported from Spain with a non-healing, ulcerated nodule on a hindleg. The presence of Leishmania species was detected by histopathology and pcr on samples of skin. The lesion was unresponsive to treatment with allopurinol for three months but the cat was treated successfully by removing the lesion surgically. The second cat had lived in both Spain and Switzerland, and had a history of recurrent skin lesions on its head and neck. A diagnosis of pemphigus foliaceus was made on the basis of histopathology, but Leishmania species serology (elisa) and pcr of skin were positive, leading to a diagnosis of a Leishmania species infection combined with pemphigus foliaceus.

Animals↗