No abstract available.
Antibodies ; Desmoglein 1 ; Desmogleins ; Pemphigus

OBJECTIVETo investigate the relationship between the levels of antidesmoglein (DSG) 1, 3 antibodies in the sera of patients with paraneoplastic pemphigus (PNP) and alopecia.

METHODSSera from PNP patients, bullous pemphigoid patients, and normal healthy subjects were collected and 2 tissue samples from 2 healthy scalps were resected. Anti-DSG 1, 3 antibodies in the sera of PNP patients were detected by enzyme-linked immunosorbent assay (ELISA). Indirect immunofluorescent assay was used to detect whether the antibodies in the sera of PNP patients binds with the follicular epithelium of normal healthy scalp.

RESULTSAnti-DSG3 autoantibody was strongly positive and anti-DSG1 weakly positive in one patient, while both two antibodies were negative in the other patient. Their sera could bind to keratinocytes and follicular epithelium in human scalp. Immunofluorescent signals were found on the intercellular epidermal cell surface and outer root sheath of the follicular epithelium. However, the immunofluorescent signals in the section incubating with serum of bullous pemphigoid were only found on basal membrane zone. No signals were found in the section incubating with normal healthy serum.

CONCLUSIONAlopecia in PNP patients are correlated with the anti-DSG3.

Adult ; Alopecia ; etiology ; immunology ; Autoantibodies ; blood ; Desmoglein 1 ; immunology ; Desmoglein 3 ; immunology ; Female ; Humans ; Male ; Paraneoplastic Syndromes ; complications ; immunology ; Pemphigus ; complications ; immunology

BACKGROUND: Pemphigus are chronic autoimmune blistering disorcers characterized by acantholysis. In addition to pemphigus vulgaris(PV), the major clinical variarts are pemphigus foliaceus(PF), paraneoplastic pemphigus(PNP) and drug-induced pemphigus(DP). Detection of pemphigus antigen is important for differential diagnosis as well as research work. Most investigators have identified pemphigus antigens by means of immunoprecipitation using metabolically radiolabeled cultured keratinocytes. However, immunorepitation is generally more expensive, hazardous and time-consuming than immunoblotting. Therefore, establishment of the immunoblotting as a standard technique for the detection of the pemphig us antigens is desirable. OBJECTIVE: To characterized pemphigus antigens by an immunobloting analysis of human epidermal extract and by indirect itnmunofluroscence study using human of cultured keratinocytes as a substraie. METHOD: We performed imrnunoblotting analysis af sera from patieiits with PV, PF, PNP and DP with human epidermal extract as a source of antigen. Indirect immunof uorescence study was also performed using human keratinocytes cultured in high or low calcium media for detection of pemphigus antigens. RESULTS: In an immunoblotting analysis, all(9/9) PV sera showed secific reactivities with a 130-KD protein and all(5/5) FF sera showed reactivities with a 150-KK protein, which is most likely desmoglein 1. Furthermore, one of nine PV serum also reacted with a 150-KD protein, which seems to be the identical antigen detected in PF. All PNP(3/3) sera showed reactivities with two protein bands, 210KD and 190KD. In our indirect imrnunofluorescence study using culltured human keratinocytes as a substrate, when keratinocytes were grown in low calcium media, no pimphigus antigens could be detected. However, when grown in high calciurn media, pemphigus vulga ris and paraneoplastic pernphigus antigens were present t the cell-cell contact areas with a puncta;e pattern, whereas pemphigus foliaceus antigen was not, presint in keratinocytes even when cultured in high calcium media. CONCLUSION: Our results suggests (1) immunoblotting analysis is a reliable technique for defining pemphigus antigen and could be a valuable tool for the differentiation of PV, PF and PNP and(2) PF antigen rnay not be expresseden cultured keratinocytes.
Acantholysis ; Blister ; Calcium ; Desmoglein 1 ; Diagnosis, Differential ; Fluorescent Antibody Technique, Indirect* ; Humans ; Immunoblotting ; Immunoprecipitation ; Keratinocytes* ; Pemphigus* ; Research Personnel

BACKGROUND: Desmogleins are transmembrane glycoproteins of the desmosome which provide mechanical strength to epithelial tissue. Desmogleins have so far, been implicated in several diseases such as pemphigus, striate palmoplantar keratoderma, 4S and squamous cell carcinomas. Skin cancer usually occurs in old age. And there are reports that the expression of desmogleins are increased in squamous cell carcinoma. However the role of desmogleins in skin aging has not yet been reported. OBJECTIVE: The purpose of this study was to investigate the expression of desmoglein 1 and 3 according to chronologic skin aging. METHODS: A total of 6 normal tissue samples from sun-protected skin of different age groups (from 34-year-old to an 84-year-old) and 1 squamous cell carcinoma tissue from a 72-year-old patient were taken. Western blotting and immunohistochemical staining were performed with anti desmoglein 1 and 3 antibodies. The expression of desmoglein 1 and 3 by Western blotting were calculated semiquantitatively by a densitometer. RESULTS: The expression of desmoglein 1 was 0.382 in the 34-year-old, 0.450 in the 45-year-old, 0.369 in the 56-year-old, 0.761 in the 65-year-old, 1.035 in the 77-year-old and 1.329 ODu/mm2 in the 84-year-old. The expression of desmoglein 3 was 0.830 in the 34-year-old, 0.984 in the 45-year-old, 1.029 in the 56-year-old, 1.534 in the 65-year-old, 1.714 in the 77-year-old and 1.878 ODu/mm2 in the 84-year-old. In immunohistochemical staining, the expression of Dsg1 increased from the basal layer to the granular layer and Dsg3 was expressed in the basal and suprabasal layers. CONCLUSION: The expression of desmoglein 1 and 3 were increased according to chronologic skin aging.
Adult ; Aged ; Aged, 80 and over ; Antibodies ; Blotting, Western ; Carcinoma, Squamous Cell ; Desmoglein 1* ; Desmoglein 3 ; Desmogleins* ; Desmosomes ; Glycoproteins ; Humans ; Keratoderma, Palmoplantar ; Middle Aged ; Pemphigus ; Skin Aging* ; Skin Neoplasms ; Skin*

BACKGROUND: Topical retinoids induce skin fragility. As corneodesmosomes are important adhesion structures in the epidermal cohesion, an effect of retinoids on corneodesmosomes has been suspected. OBJECTIVE: The aim of this study was to investigate the effect of retinoid on the expression of corneodesmosomal components including desmoglein (DSG) 1, desmocollin (DSC) 1, corneodesmosin (CDSN) and kallikrein (KLK)s. METHODS: 2% all-trans-retinol or ethanol was applied to the back of hairless mice for five days, and the structure of the stratum corneum was examined by transmission electron microscopy. The cultured human keratinocytes were treated with all-trans-retinoic acid (RA) in low or high calcium media for 24 hours. RESULTS: Topical retinol increased corneocyte detachment and degradation of corneodesmosomes. RA significantly decreased DSG1 and DSC1 expression at the mRNA and protein levels in keratinocytes that were cultured in both low- and high-calcium media. On the other hand, CDSN mRNA levels did not decrease in low-calcium media or increase in high-calcium media after RA treatment. KLK5 and KLK7 expression did not increase after RA treatment. CONCLUSION: Our results indicate that DSG1 and DSC1 downregulation by RA could be related to the increased degradation of corneodesmosomes and consequent desquamation induced by retinoids.
Animals ; Calcium ; Desmoglein 1 ; Desmogleins ; Down-Regulation ; Ethanol ; Hand ; Humans ; Kallikreins ; Keratinocytes ; Mice ; Mice, Hairless ; Microscopy, Electron, Transmission ; Retinoids ; RNA, Messenger ; Skin ; Tretinoin ; Vitamin A

Paraneoplastic pemphigus is a rare, life-threatening autoimmune mucocutaneous blistering disease associated with underlying neoplasia, commonly lymphoproliferative tumors. Herein we report a case of paraneoplastic pemphigus with a unique autoantibody profile associated with a malignant thymoma. A 56-year-old female patient presented with relapsing oral ulcerations accompanied by erythematous papules and patches on her extremities for 2 months. Skin and mucosal biopsies identified interface dermatitis with lichenoid lymphocytic infiltration in the upper dermis. Immunoblotting and enzyme-linked immunosorbent assays revealed that the patient had multiple autoantibodies against desmoglein 1, desmocollin 1, 2, 3, laminin gamma-1, envoplakin, and periplakin. The skin lesions completely healed following thymectomy and systemic corticosteroid therapy, but the oral ulcerations persisted through a follow-up period of over 2 years.
Autoantibodies ; Biopsy ; Blister ; Dermatitis ; Dermis ; Desmoglein 1 ; Enzyme-Linked Immunosorbent Assay ; Extremities ; Female ; Follow-Up Studies ; Humans ; Immunoblotting ; Laminin ; Middle Aged ; Oral Ulcer* ; Paraneoplastic Syndromes ; Pemphigus* ; Skin ; Thymectomy* ; Thymoma*

OBJECTIVE: To examine the effect of HPV-16 E6 expression on the transcription of cellular genes, we used cDNA microarray in HPV-16 E6 transfected stable cancer cell lines. METHODS: Using cDNA microarray consisting of 1,024 genes, we have performed a systematic characterization of gene expression in A549E6 human lung adenocarcinoma and RC10.1 human colon adenocarcinoma cell lines stably expressing HPV-16 E6 gene. The up-regulated and down-regulated genes were classified into the different functional categories; oncogenes, apoptosis, cell cycle, signal transduction, gene regulation, immune response, cell adhesion, protein transport, metabolism, redox control and angiogenesis. RESULTS: Among 1,024 known genes and ESTs (expressed sequence tags) tested, we found 27 up- regulated and 43 down-regulated genes in A549E6 (HPV-16 E6) compared to A549. The major up-regulated genes were as follows. GTPase-activating protein Rho 4, transcription factor D2, IKAROS, integrin-alpha 6, cadherin 11, ephrin-beta 2, RAN binding protein 2, branched-chain amino transferase 2. The major down-regulated genes were as follows. K-ras 2, CDC (cell division cycle) 37, CDC16, CDC7L1, IRF3, interferon-gamma-inducible protein 30, cadherin 6, desmoglein 1, desmocollin 2, endothelin 2. Also, we found 48 up-regulated and 34 down-regulated genes in RC10.1 (HPV-16 E6) compared to RKO. The major up-regulated genes were as follows. Colon cancer familial nonpolyposis type 1 (COCA 1), Bcl 2, jagged 1, MAP2K6, E2F1, ephrin receptor-beta 2, ephrin-beta 2, desmoglein 1, transforming growth factor-beta 3. The major down-regulated genes were as follows. KIT, Rad51C, Bcl 2 antagonist killer 1, STAT 4, epidermal growth factor receptor, high mobility group protein 2, cadherin 11, cadherin 12, cadherin 3, integrin-alpha 1, intergrin-alpha 8, chromosome segregation 1-like. CONCLUSION: Various expression patterns of cellular genes by HPV-16 E6 could be wholy grasped and classified into different functional groups using both cell line system stably expressed HPV-16 E6 and cDNA microarray analysis. These analysis methods must be helpful to understand multiple effects of a specific gene on cellular genes in a short period.
Adenocarcinoma ; Apoptosis ; Cadherins ; Carrier Proteins ; Cell Adhesion ; Cell Cycle ; Cell Line ; Centers for Disease Control and Prevention (U.S.) ; Chromosome Segregation ; Colon ; Colonic Neoplasms ; Desmoglein 1 ; DNA, Complementary* ; Endothelin-2 ; Expressed Sequence Tags ; Gene Expression* ; GTPase-Activating Proteins ; Hand Strength ; Human papillomavirus 16* ; Humans ; Lung ; Metabolism ; Oligonucleotide Array Sequence Analysis* ; Oncogenes ; Oxidation-Reduction ; Protein Transport ; Receptor, Epidermal Growth Factor ; Signal Transduction ; Transcription Factors ; Transferases