Associations of anti-desmoglein antibodies with clinical phenotypes and disease activity in pemphigus patients and their change patterns / 中华皮肤科杂志

Objective To investigate associations of anti-desmoglein (Dsg1 and Dsg3) antibodies detected by enzyme-linked immunosorbent assay (ELISA) with clinical phenotypes and disease activity in pemphigus patients,and to explore their change patterns.Methods A total of 111 patients with pemphigus were enrolled from Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College between January 2015 and January 2018.ELISA was performed to detect serum levels of anti-Dsg1 and anti-Dsg3 antibodies in these patients with different clinical types of pemphigus at different stages,including onset stage,control stage (no new erythema or vesicles occurred in the last 2 or more weeks,and primary lesions began to regress),maintenance stage (the condition had been stable for ≥ 1 month,and treatment was maintained with a low dose of glucocorticoids [prednisone equivalent of ＜ 15 mg/d]),and recurrence stage,and the change patterns of serum levels of anti-Dsg1 and anti-Dsg3 antibodies were analyzed.Statistical analysis was carried out with SPSS 22 software by using oneway analysis of variance for the comparison among groups,and least significant difference (LSD)-t test for multiple comparisons.Results At the disease onset stage,control stage,maintenance stage and recurrence stage,92,53,33,and 9 patients respectively completed the detection.Among the 92 patients with initial onset of pemphigus,the positive rates of anti-Dsg1 and anti-Dsg3 antibodies were 100％ and 2.77％ respectively in 36 patients with pemphigus foliaceus,20％ and 80％ respectively in 10 with mucosaldominant pemphigus vulgaris,and 97.82％,95.65％ respectively in 46 with mucocutaneous pemphigus vulgaris.The serum levels of anti-Dsg1 antibodies in the patients with pemphigus foliaceus significantly differed among the disease onset stage,control stage,maintenance stage and recurrence stage (137.43 ±77.74,13.94 ± 14.81,21.50 ± 58.33,121.13 ± 86.89 U/ml,respectively),the serum levels of anti-Dsg3 antibodies in the patients with mucosal-dominant pemphigus vulgaris also significantly differed among the above clinical stages (125.61 ± 94.81,34.5 ± 16.26,0.6,258 U/ml,respectively),and the serum levels of anti-Dsg1 and anti-Dsg3 antibodies in patients with mucocutaneous pemphigus vulgaris both significantly differed among the above clinical stages(anti-Dsg1 antibody:115.39 ± 70.62,15.74 ± 25.10,3.62 ± 12.09,78.60 ± 92.25 U/ml,respectively;anti-Dsg3 antibody:137.98 ± 81.25,58.14 ± 63.46,29.26 ± 64.70,136.9 ± 101.47 U/ml,respectively).Additionally,the serum levels of anti-Dsg1 antibodies in the patients with pemphigus foliaceus,as well as the serum levels of anti-Dsg3 antibodies in the patients with mucosaldominant pemphigus vulgaris and those with mucocutaneous pemphigus vulgaris,were both significantly lower at the disease control stage and maintenance stage than at the disease onset stage and recurrence stage (all P ＜ 0.05).During the treatment,epitope spreading occurred in 2 patients,and high-titer anti-Dsg antibodies were observed in 4 patients at the stable stage.Conclusion Anti-Dsg antibody spectrum is associated with clinical phenotypes of pemphigus,and its serum levels measured by ELISA can be applied to disease activity monitoring and evaluation of therapeutic efficacy.

A study of prevalence of autoantibodies in patients with lichen planus from Mumbai, India

Background: Lichen planus is a common chronically relapsing autoimmune skin condition with poorly understood etiology. Apart from cellular immunity, presence of various antibodies has been hypothesized. Various studies have found the presence of serum anti-nuclear antibody, anti-mitochondrial antibody, anti-desmoglein 1 and 3 antibodies, anti-keratinocyte antibody and anti-thyroglobulin antibody in patients of cutaneous and oral lichen planus. Aim: To study the prevalence of autoantibodies and the clinical spectrum of disease in an Indian patient subpopulation with lichen planus. Methods: A cross-sectional epidemiological study comprising 100 lichen planus patients was conducted in the dermatology outpatient department of Seth G.S Medical College and King Edward Memorial Hospital, Mumbai, Maharashtra, India. Serum concentrations of circulating anti-nuclear antibodies, anti-desmoglein 1 antibody, anti-desmoglein 3 antibody, anti-keratinocyte antibodies, anti-mitochondrial antibodies and anti-thyroglobulin antibodies were determined by indirect immunofluorescence. Pairs of groups were compared using “Student's t-test” for normally distributed continuous data. The “χ2-test” was used for the categorical variables as needed. Statistical significance was set at P < 0.05. Results: It was found that 65 (65%) patients showed the presence of at least one of the six autoantibodies that we studied, while 35 (35%) tested negative for all six of them. Positivity of anti-keratinocyte antibody in 26 (26%), anti-nuclear antibody in 22 (22%), anti-desmoglein 1 antibody in 19 (19%), anti-desmoglein 3 antibody in 16 (16%), anti-mitochondrial antibody in 9 (9%) and anti-thyroglobulin antibody in 6 (6%) patients was detected. It was observed that 55 (71.4%) patients of cutaneous lichen planus, 6 (46.1%) patients of mucosal lichen planus and 4 (40%) patients of cutaneous and mucosal lichen planus overlap showed presence of at least one autoantibody. Conclusion: This study provides the serological parameters of a population of lichen planus from western India. Presence of autoantibodies in lichen planus suggests the possible role of humoral immunity in lichen planus. Identifying antibodies linked to lichen planus may help in identifying suitable diagnostic tests and therapeutic targets. Well-controlled studies with larger sample size are the need of the hour to confirm the role of humoral immunity in lichen planus. Limitations: Studies with a larger number of patients as well as controls should be undertaken to further evaluate the role of autoantibodies in lichen planus.

Expression and specific antibody-producing capacity of B lymphocytes infiltrating in lesions of patients with pemphigus vulgaris / 中华皮肤科杂志

Objective To evaluate the specific antibody-producing capacity of locally infiltrating B lymphocytes in lesions of patients with pemphigus vulgaris (PV).Methods Totally,35 patients with PV and 22 healthy controls were enrolled into this study.Skin tissues were resected from blisters or erosions of the patients with PV,and from normal skin of healthy controls.Then,mononuclear cells were isolated from these skin tissues.Flow cytometry was performed to determine the percentages of lymphocytes,CD 19+ B lymphocytes,and desmoglein (Dsg)1-and Dsg3-specific CD19+ B lymphocytes.B lymphocytes isolated from the lesional skin of patients with PV were cultured in vitro.Enzyme-linked immunosorbent assay (ELISA) was conducted to determine titers of anti-Dsg1 and anti-Dsg3 antibodies in the cell culture supernatant.Receiver operating characteristic (ROC) curve analysis was conducted to calculate positive rates of anti-Dsg1 and anti-Dsg3 antibodies.Results The percentages of lymphocytes (17.95％ ± 3.85％) and CD19+ B lymphocytes (4.27％ ± 1.13％) were significantly higher in the lesional skin of PV patients than in the normal skin of healthy controls (7.83％ ± 1.29％,0.61％ ± 0.31％ respectively;t =2.49,U =13.00 respectively,both P ＜ 0.05).Among the CD19+ B lymphocytes in the lesional skin of PV patients,the percentage of CD19qgG+ B cells was (38.33 ± 5.56)％,and percentages of Dsg1-and Dsg3-specific CD19+ B lymphocytes were 12.87％ ± 1.267％ and 10.42％ ± 1.243％ respectively.After the in vitro culture for 6 days,the titers of anti-Dsg1 and anti-Dsg3 antibodies in the cell culture supematant were (4.89 ± 1.56) U/ml and (35.45 ± 13.03) U/ml respectively,with their positive rates being 85％ (17/20)and 95％ (19/20) respectively.Conclusion There are Dsg1-and Dsg3-specific B lymphocytes aggregating in the lesional skin of patients with PV,which can produce anti-Dsg1 and anti-Dsg3 antibodies after in vitro culture.

Pathological features of ectopic lymphoid structure in skin lesions of patients with pemphigus / 中华皮肤科杂志

Objective To investigate pathological features of infiltrating lymphocytes in skin lesions of patients with pemphigus,and to analyze their correlation with titers of anti-desmoglein (Dsg) 1 and anti-Dsg3 antibodies in peripheral blood.Methods A retrospective pathological analysis was performed in 93 patients with pemphigus vulgaris or pemphigus foliaceus,who visited the Department of Dermatology of Ruijin Hospital affiliated to Shanghai Jiao Tong University School of Medicine between 2014 and 2016.For each HE-stained section,the total number of lymphocytes per × 50 microscopic field was counted,and defined as lymphocyte density index.Enzyme-linked immunosorbent assay (ELISA) was conducted to determine the serum titers of anti-Dsg1 and anti-Dsg3 antibodies in the patients with pemphigus.The correlations between the lymphocyte density index and titers of anti-Dsg1 and anti-Dsg3 antibodies were analyzed.Immunohistochemical staining was performed in lesional skin samples from 8 patients with pemphigus vulgaris and 8 patients with pemphigus foliaceus,so as to analyze the distribution of CD3+ T cells,CD20+ B cells and CD138+ plasma cells.Results Of the 93 pathological sections,93 (100.00％) showed Grade1 lymphocyte aggregates,64 (68.09％) showed Grade 2 lymphocyte aggregates,and 10 (10.64％) showed Grade 3 lymphocyte aggregates,and the 56 cases of pemphigus vulgaris and 37 of pemphigus foliaceus showed the similar proportion of grade 1,2 and 3 lymphocyte aggregates.There was also no significant difference in the lymphocyte density index between patients with pemphigus vulgaris and pemphigus foliaceus (P ＞ 0.05),and the lymphocyte density index was uncorrelated with the serum titers of anti-Dsg1 and anti-Dsg3 antibodies in patients with pemphigus.Of the 16 cases of pemphigus,CD3+ T cells were found in all cases,CD20+ B cells in 15,and CD138+ plasma cells in 12.Of 16 sections,all showed a large amount of CD3+ T cells in Grade 1-3 lymphocyte aggregates,while lymphocyte aggregates containing CD20+ B cells and CD138+ plasma cells were found in 52.80％ ± 5.78％ and 34.59％ ± 7.42％ of sections respectively.No significant differences in the distribution of CD3+ T cells,CD20+ B cells,CD138+ plasma cells were found between the 8 cases of pemphigus vulgaris and 8 cases of pemphigus foliaceus.Conclusion Different degrees of lymphocyte infiltration generally exist in skin lesions of patients with pemphigus,which may form ectopic lymphoid structures and contribute to the development and aggravation of pemphigus skin lesions.

Locations of acantholysis in 71 cases of pemphigus / 中华皮肤科杂志

Objective To analyze locations of acantholysis in pemphigus vulgaris (PV) and pemphigus foliaceus (PF),so as to explain why acantholysis in pemphigus occurs in different locations of the epidermis.Methods Clinical data,histopathological and immunopathological findings,and pemphigus antibody level values were collected from 43 patients with PV and 28 with PF,and retrospectively analyzed.Results Of the 43 patients with PV,35 showed acantholysis in the upper basal layer,8 in the middle-to-upper epidermis.Of the 28 patients with PF,25 showed acantholysis in the granular layer and the upper prickle cell layer,3 in the middle-to-lower epidermis.Patients with PF showed significantly higher levels of anti-Desmoglein 1 (Dsg1) antibody compared with patients with PV (P =0.047).However,there were no significant differences in the levels of anti-Dsg1 and anti-Dsg3 antibodies between PV patients who had acantholysis in the middle-to-upper epidermis and those in the upper basal layer.Conclusion Histopathological examinations of PV and PF lesions show that acantholysis can occur in the middle-to-upper epidermis,as well as in the middle-to-lower epidermis,and locations of acantholysis may be associated with levels of anti-Dsg1 and anti-Dsg3 antibodies.

Analysis on relationship between serum anti-desmoglein antibodies and disease in patients with pemphigus / 重庆医学

Objective To investigate the application value of ELISA for detecting the serum anti desmoglein (Dsg) 1 and Dsg 3 in the diagnosis and treatment of pemphigus .Methods Forty‐seven patients with pemphigus in our hospital from January to De‐cember 2014 were selected as the observation group and contemporaneous 52 patients with excluding pemphigus were selected as the control group .The Dsg antibodies were detected by using indirect immunofluorescence method and Dsg 1 and Dsg3 were deter‐mined by ELISA ;their correlation with pemphigus characteristics was analyzed .Results The sensitivity and specificity of ELISA for detecting anti‐Dsg antibodies were 95 .74% and 92 .31% respectively ,while which of IIF were 93 .62% and 86 .54% respective‐ly ,showing no statistically significant difference between the two test methods (P>0 .05) .In 30 cases of pemphigus vulgaris ,16 ca‐ses (16/30) were positive Dsg1 and Dsg 3 ,8 cases of pemphigus erythematosus and 5 cases pemphigus foliaceus were positive Dsg1 only ,and 2 cases of pemphigus vegetans were both positive Dsgl and Dsg3 .The Dsgl and Dsg3 titers of pemphigus vulgaris and pemphigus vegetans were 130 .85 ± 86 and 112 .30 ± 85 .05 ,respectively ,and the disease activity score was (5 .10 ± 1 .86) points ,the correlation coefficient(r)=0 .476(P=0 .008) ,r=0 .816(P=0 .001) ,respectively .The Dsgl titer of pemphigus erythematosus and pemphigus foliaceus were 142 .59 ± 78 .52 ,and the disease activity score was (2 .77 ± 0 .92) points(r=0 .800 ,P=0 .001) .Conclu‐sion ELISA for detecting Dsg1 and Dsg3 has high sensitivity and specificity ,and is conducive to the diagnosis of pemphigus and e‐valuation of disease severity .

Mechanisms underlying the reversal of acantholysis in pemphigus by a cholinergic receptor agonist / 中华皮肤科杂志

Objective To evaluate the reversal effect of a cholinergic receptor agonist on acantholysis in pemphigus,and to investigate its mechanism.Methods Human HaCaT keratinocytes were co-cultured with pemphigus vulgaris immunoglobulin G (PV-IgG) to establish a cell model of pemphigus,then classified into two groups to be incubated with the cholinergic receptor agonist carbachol for 12 hours (test group) or remain untreated (control group).Cell dissociation assay was performed to quantitatively estimate the reversal effect of carbachol on acantholysis,and immunofluorescence assay to qualitatively assess the changes of desmosomal proteins.Radio-immunoprecipitation assay (RIPA) lysis buffer and Triton X-100 were used to lyse HaCaT cells to obtain total proteins and cytoplasmic proteins,and Western blot was conducted to determine the expression levels of adhesion-related proteins desmoglein 3 (Dsg3) and plakoglobin (PG) on the surface of HaCaT cells,as well as the phosphorylation levels of p38 mitogen activated protein kinase (p38 MAPK) and epidermal growth factor receptor (EGFR) at different time points.Quantitative polymerase chain reaction (qPCR) was performed to detect the mRNA expressions of the above surface proteins,and coimmunoprecipitation assay to qualitatively evaluate the interaction between Dsg3 and PG.Results The number of cell debris was significantly lower in the test group than in the control group (18.67 ± 2.52 vs.46.67 ± 2.03,t =11.22,P＜0.01).Immunofluorescence assay showed that carbachol could reverse the internalization of desmosomal molecules induced by PV-IgG.In the pemphigus cell model,the levels of total Dsg3 and PG as well as non-desmosomal Dsg3 were decreased,while the level of non-desmosomal PG increased,and the interaction between Dsg3 and PG was attenuated.When the pemphigus cell model was co-cultured with carbachol,these above changes were reversed.Carbachol also increased the mRNA levels (expressed as 2-△△Ct) of Dsg3 and PG from 1.428 ± 0.215 and 1.563 ± 0.247 in the control group to 4.974 ± 0.948 (t =3.65,P =0.01) and 13.420 ± 1.715 (t =6.85,P ＜ 0.01) in the test group respectively.In phosphorylation assay,carbachol inhibited the phosphorylation of EGFR,but had no significant effect on that of p38 MAPK.Conclusions The cholinergic receptor agonist carbachol can reverse acantholysis in pemphigus,likely by inhibiting the internalization of Dsg3 and PG,enhancing their expressions and interaction,and suppressing the phosphorylation of the key signaling molecule for acantholysis,EGFR.

The value of trichoscopy in the differential diagnosis of scalp lesions in pemphigus vulgaris and pemphigus foliaceus

BACKGROUND: Trichoscopy is becoming increasingly popular in diagnosing hair and scalp diseases. Scalp involvement in pemphigus is common. The scalp may be the first or only site of clinical manifestation of the disease. OBJECTIVE: The aim of this study was to analyze whether trichoscopy may be useful in aiding differential diagnosis of scalp lesions in patients with pemphigus vulgaris and pemphigus foliaceus. METHODS: Trichoscopy was performed in 19 patients with scalp lesions in the course of pemphigus (9 patients with pemphigus vulgaris and 10 with pemphigus foliaceus). In all patients, the diagnosis of scalp pemphigus was confirmed by histopathology. The working magnification was 20-fold and 70-fold. RESULTS: The most frequently observed trichoscopy features of pemphigus lesions were: extravasations (18/19; 94.7%) and yellow hemorrhagic crusts (11/19; 57.9%). Yellow dots with whitish halo were observed in 6/19 (31.6%) patients with pemphigus. White polygonal structures were observed in pemphigus foliaceus (6/10; 60%), but not in pemphigus vulgaris. Vascular abnormalities were more frequent in pemphigus vulgaris, when compared to pemphigus foliaceus, and were associated with a severe course of disease. Linear serpentine vessels were the most frequent vascular abnormality in patients with pemphigus vulgaris and pemphigus foliaceus (77.8% and 30%, respectively). CONCLUSION: Trichoscopy may serve as a useful supplementary method in the differential diagnosis of pemphigus, especially in cases of desquamative or exudative lesions limited to the scalp. Extravasations, yellow hemorrhagic crusts, yellow dots with whitish halo, white polygonal structures and linear serpentine vessels are trichoscopy features which may suggest the diagnosis of pemphigus. .

Expressions of desmoglein 1 and desmoglein 3 in primary keratinocytes and two keratinocyte cell lines HaCaT and A431 / 中华皮肤科杂志

Objective To detect the mRNA and protein expressions of desmoglein 1 (DSG1) and DSG3 in different types of keratinocytes (KCs).Methods Two keratinocyte cell lines HaCaT and A431,as well as primary keratinocytes from human abdomenal skin served as the object of this study.Direct immunofluorescence assay was performed to observe and quantify the expressions of DSG1 and DSG3,and quantitative PCR (qPCR) to determine the mRNA expressions of DSG1 and DSG3,in these cells.Results Both DSG1 and DSG3 were expressed in all the three types of keratinocytes,and the fluorescence intensity of DSG1 and DSG3 in HaCaT cells was higher than that in primary keratinocytes but lower than that in A431 cells.Similarly,all the keratinocytes expressed DSG1 and DSG3 mRNA,with the relative expression levels of DSG1 and DSG3 mRNA in primary keratinocytes being 291.7％ and 237.4％ of those in HaCaT cells respectively (both P ＜ 0.01),and those in A431 cells being 0.1％ and 18.8％ of those in HaCaT cells respectively (both P ＜ 0.05).Conclusions HaCaT cells,A431 cells and primary keratinocytes all can be used for the study of DSG1 and DSG3,of which,A431 cells show the strongest expressions of DSG1 and DSG3,and primary keratinocytes display the highest expressions of DSG1 and DSG3 mRNAs.

Proliferation of desmoglein 3-stimulated T lymphocytes from patients with pemphigus vulgaris / 中华皮肤科杂志

Objective To investigate the impact of desmoglein 3 (Dsg3) on the proliferation of peripheral T lymphocytes from patients with pemphigus vulgaris (PV).Methods Peripheral blood mononuclear cells (PBMCs) were obtained from 12 patients with PV and 22 normal human controls,cultured with or without the presence of Dsg3 or phytohemagglutinin for 3 days.Flow cytometry was performed to detect the changes of T-lymphocyte subsets in PBMCs stimulated with Dsg3 and proliferation of T-lymphocytes.Results In patients with PV,the percentage of Th2 and Th1 cells was 12.17%±5.32% and 4.08%±1.50%,respectively in Dsg3stimulated PBMCs,9.84%±5.41% and 3.91%±1.38%,respectively in non-stimulated PBMCs.Increased percentage of Th2 cells was observed in Dsg3-stimulated and non-stimulated PBMCs from patients with PV compared with those from normal human controls (both P<0.05).After stimulation with Dsg3,there was a significant proliferation of T cells from patients,and the proliferation rate of CD4+T cells was 4.65%±3.28%,which was significantly higher than that from normal controls(P<0.05).Conclusion Dsg3 can induce the specific proliferation of CD4+T cells,especially Th2 type CD+4 T cells,from patients with PV.

Immunohistochemical Study on the Expression of Desmoglein-3 in Fetal Skin Development / 대한피부과학회지

BACKGROUND: Desmogleins are calcium-dependent transmembrane glycoproteins of the desmosome that form an import component of the junction complexes of epithelial cells. Desmogleins are involved in maintaining the structural integrity of tissues. So far, four different desmogleins (Dsg1, Dsg2, Dsg3 and Dsg4) have been identified. OBJECTIVE: The purpose of this study was to observe the distribution pattern of desmoglein-3 in the fetal skin during development. METHODS: Skin was obtained from the sole, scalp and lip of 34 fetuses that ranged in age from 10 to 39 weeks of gestational age. Immunohistochemical staining was performed on the paraffin embedded tissue using anti-human monoclonal antibody against the desmoglein-3. RESULTS: The expression of desmoglein-3 in the epidermis appeared in the basal layer of the sole at the 10th week of gestation age. Thereafter, a stronger expression was noted in the middle layer of the sole and scalp epidermis. The basal layer had a stronger expression than did the other layers of the epidermis, followed by the middle and superficial layers. A stronger expression of desmoglein-3 in hair was noted in the outer root sheath, the bulge cells and the eccrine duct cells. The expression of desmoglein-3 in the lip mucosa was strong in both the basal and middle layers, while the skin side of the mucosa showed a stronger expression in basal layer. CONCLUSION: These results suggested that desmoglein-3 plays an important role in the development and differentiation of the epidermis and skin adnexa in the fetal stage, and especially in basal and suprabasal layers.

Expression of N-terminal truncated desmoglein 3 (Delta NDg3) in epidermis and its role in keratinocyte differentiation

During a search for keratinocyte differentiation-related genes, we obtained a cDNA fragment from the 5'-untranslated region of a previously identified splicing variant of desmoglein 3 (Dg3). This transcript encodes a protein of 282 amino acids, which corresponds to the N-terminal truncated intracellular domain of Dg3 (Delta NDg3). Northern blot analysis detected a 4.6-kb transcript matching the predicted size of Delta NDg3 mRNA, and Western blot analysis with an antibody raised against the Dg3 C-terminus (H-145) detected a 31-kDa protein. Increased Delta NDg3 expression was observed in differentiating keratinocytes by RT-PCR and Western blot analysis, suggesting that Delta NDg3 is indeed a differentiation-related gene product. In immunohistochemical studies of normal and pathologic tissues, H-145 antibody detected the protein in the cytoplasm of suprabasal layer cells, whereas an antibody directed against the N-terminal region of Dg3 (AF1720) reacted with a membrane protein in the basal layer. In addition, Delta NDg3 transcript and protein were upregulated in psoriatic epidermis, and protein expression appeared to increase in epidermal tumors including Bowen's disease and squamous cell carcinoma. Moreover, overexpression of Delta NDg3 led to increased migration and weakening of cell adhesion. These results suggest that Delta NDg3 have a role in keratinocyte differentiation, and that may be related with tumorigenesis of epithelial origin.

Selection of mimotopes of pemphigus vulgaris antigen from a phage-displayed random nonapeptide library / 中华皮肤科杂志

Objective To screen the mimotopes ofpemphigus vulgaris (PV) antigen, desmoglein3 (Dsg3) with a phage-displayed random nonapeptide library, so as to update the knowledge on the patho-genesis of PV. Methods Recombinant fusion protein of extracellular domain 1-2 (EC1-2) of Dsg3 and glutathione transferase was expressed by E.coli BL21, and used to purify polyclonal autoantibody binding to recombinant EC 1-2 from the sera of patients with PV. Then, selected autoantibody was applied as a ligand for biopanning of a phage-displayed linear random nonapeptide library and circular random nonapeptide library. Monoclonal phages were selected by immunoscreening and tested with ELISA and competitive ELISA. Results After two rounds ofbiopanning, a population ofpeptide-displaying phages binding to autoan- tidody were highly enriched. Sixty individual phage clones selected by immunosereening were further sub-jected to screening with ELISA and competitive ELISA. Finally, three positive phage clones were obtained. As shown by ELISA and competitive ELISA, they reacted with serum from patients with PV but not with that from normal human controls, and blocked the interaction between patients' sera and recombinant fusion protein of EC1-2. Conclusion Three mimotopes closely associated with PV antigen were successfully selected from a phage-displayed random nonapeptide library.

The Expressions of Desmoglein 1 and 3 according to Chronologic Skin Aging / 대한피부과학회지

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.