Objective::Pemphigus vulgaris (PV) is a severe autoimmune skin disease, and the role of acetylcholine receptor (AChR) in PV is still unclear. This study aimed to investigate the effects of M3 AChR and α9 AChR interference on acantholysis of keratinocytes, which may provide a deeper understanding of PV pathogenesis.Methods::This was an in vitro experimental study designed to explore the roles of M3 AChR and α9 AChR in PV pathogenesis. Sera from patients with PV and controls were collected from Hospital for Skin Diseases (Institute of Dermatology), Chinese Academy of Medical Sciences and Peking Union Medical College from January 1, 2016, to December 30, 2016, and were used to extract PV-immunoglobulin G (IgG). RNA interference was used to silence the M3 AChR gene or α9 AChR gene in HaCaT cells, after which the cells were treated with IgG from PV patients or controls. Changes in the adhesion of HaCaT cells were observed using a cell dissociation assay. Immunofluorescence assay was used to detect the staining patterns of desmosome proteins. Western blot analysis was performed to detect cytoskeleton- or noncytoskeleton-associated desmosome protein levels. The interactions of desmoglein-3 and plakoglobin were qualitatively detected using co-immunoprecipitation, and the p38 MAPK and epidermal growth factor receptor (EGFR) phosphorylation levels were examined. Two-tailed Student’s t-test was used to compare the data of the two groups. Results::With M3 AChR silencing, the HaCaT cells treated with PV-IgG-showed severe acantholysis compared to those with controls’ treatment ( P < 0.001); levels of cytoskeleton-associated desmoglein and plakoglobin were significantly reduced (both P < 0.001), whereas the phosphorylation of EGFR increased ( P = 0.001). In contrast, α9 AChR-silenced HaCaT cells exhibited fewer cell fragments than control HaCaT cells without α9 AChR silencing co-cultured with PV-IgG ( P < 0.001). The co-culture of α9 AChR-silenced cells with PV-IgG suppressed the internalization of desmoglein, and increased levels of cytoskeleton-associated desmoglein-3 and PG ( P < 0.001 and P= 0.002), and a close interaction between desmoglein-3 and plakoglobin was observed by immunoprecipitation ( P = 0.002). The phosphorylation levels of p38 MAPK and EGFR were suppressed by α9 AChR silencing (both P < 0.001). Conclusion::The findings of this study provided evidence that M3 AChR and α9 AChR may play roles in the pathogenesis of PV, demonstrating that M3 AChR exerts a protective role, whereas α9 AChR plays a pro-pathogenic effect in PV-IgG-induced acantholysis.

Despite the recognized association between bullous pemphigoid (BP) and psoriasis, the clinical and immunological profiles, and inflammation patterns of coexistence remain undefined. We therefore conducted a retrospective cohort study of 140 BP patients without psoriasis (BP alone group) and 24 BP patients with comorbid psoriasis (BP-PsO group) to characterise these features. Average age of BP onset was significantly lower in the BP-PsO group, compared with the BP alone group (median [IQR]: 67.00 [58.00-75.75] years vs. 75.00 [64.00-82.00] years) (p = 0.021). In the BP-PsO group, 21 patients (87.5%) had coexisting active psoriatic plaques and BP lesions. The overall disease activity of BP was comparable between the BP-PsO and BP alone groups, both showing typical BP immunological features. Serum IL-17A level was significantly elevated in the BP-PsO group (median [IQR]: 18.29 [10.39-43.50] pg/mL), compared with the BP alone group (median [IQR]: 10.36 [9.16-12.11] pg/mL) and psoriasis alone groups (median [IQR]: 11.65 [10.10-12.78] pg/mL), and correlated with disease activity. Flow cytometry confirmed enhanced IL-17A response in peripheral blood mononuclear cells from the BP-PsO group, with an elevated proportion of CD4+ IL-17A+ cells and IL-17A+ T follicular helper cells versus corresponding controls. IL-13 was comparably elevated in both the BP-PsO and BP alone groups, relative to psoriasis alone group and healthy controls. In a representative case, inhibition of IL-17A led to concurrent remission of both diseases. BP with psoriasis shares foundational features with idiopathic BP, but represents a distinct clinical entity characterised by a predominant IL-17A signature, highlighting its potential as a therapeutic target.

BACKGROUND:Anti-p200 pemphigoid is a rare autoimmune blistering disorder with limited data from China. This retrospective study aimed to investigate the clinical and serological characteristics of anti-p200 pemphigoid to enhance disease understanding. METHODS:We analysed 86 confirmed anti-p200 pemphigoid patients, evaluating their clinical manifestations, histopathological findings, and immunoserological profiles. RESULTS:The patients had a mean onset age of 56.92 ± 18.59 years and a male-to-female ratio of 2.31:1. The clinical presentation is highly heterogeneous, mimicking classic BP (57/86, 66.28%), and four cases (4/52, 7.69%) had concurrent psoriasis. Subepidermal blistering with variable dermal infiltrates was observed in 68 cases (68/70, 97.14%): neutrophil-predominant (24.89%), mixed neutrophilic/eosinophilic (31.43%), or eosinophil-predominant (27.14%). The positive rates of DIF, ss-IIF, IB with dermal extract, and IB with laminin γ1 C-terminal domain (LNγ1C) were 94.29% (66/70), 80.23% (69/86), 100% (86/86), and 64.29% (45/70), respectively. CONCLUSIONS:Anti-p200 pemphigoid closely resembles BP clinically but exhibits distinct immunopathological features. The partial reactivity to laminin γ1 (LNγ1) implies antigenic heterogeneity, warranting further investigation.

BACKGROUND:Anti-p200 pemphigoid is a rare autoimmune subepidermal blistering disease. Although the phenomenon of epitope spreading has been reported to be common in anti-p200 pemphigoid, the association between its clinical and immunoserological features has yet to be elucidated. OBJECTIVES:Our aim was to compare the clinical and immunoserological characteristics of anti-p200 pemphigoid patients with and without epitope spreading. METHODS:We performed a retrospective cohort study encompassing 30 patients with anti-p200 pemphigoid between January 2015 and December 2022. The clinical and immunoserological characteristics of anti-p200 pemphigoid were analyzed using combined immunoserological assays. RESULTS:Epitope spreading was observed in 11 of 30 patients (36.7%) with anti-p200 pemphigoid. Compared with patients in the non-epitope spreading group, patients in the epitope spreading group showed more heterogeneous clinical presentations (P = 0.018), a higher proportion of mucosal involvement (P = 0.003), higher Bullous Pemphigoid Disease Area Index (BPDAI) scores for skin erosions/blisters (P = 0.018), mucosal erosions/blisters (P = 0.001), activity (P = 0.017) and total scores (P = 0.022), and required a higher initial dose of prednisone for disease control (P = 0.040). CONCLUSIONS:This study supported the idea that anti-p200 pemphigoid was prone to epitope spreading. Anti-p200 pemphigoid patients with epitope spreading are more likely to present heterogeneous clinical phenotypes, frequent mucosal involvement, and a more severe and recalcitrant disease course.

Bullous pemphigoid (BP) is a complex inflammatory process with elevated levels of autoantibodies, eosinophils, neutrophils, and various cytokines. Hematological inflammatory biomarkers can reflect inflammatory state in various diseases. Up to now, the correlations of hematological inflammatory biomarkers and disease activity of BP remain unknown. The purpose of this study was to clarify the associations between hematological inflammatory biomarkers and disease activity of BP. The levels of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), platelet-to-neutrophil ratio (PNR) and mean platelet volume (MPV) of 36 untreated BP patients and 45 age and gender matched healthy controls were detected by routine blood tests. The correlations between hematological inflammatory markers and clinical characteristics of BP were statistically analyzed. The Bullous Pemphigoid Disease Area Index (BPDAI) was used to measure disease activity of BP. The mean levels of NLR, PLR, PNR and MPV in 36 untreated BP patients were 3.9, 157.9, 45.7 and 9.4 fl, respectively. Increased NLR (p < 0.001), PLR (p < 0.01), and MPV (p < 0.001) but decreased PNR (p < 0.001) were observed in BP patients when compared with healthy controls. In BP patients, the levels of NLR were positively correlated to BPDAI Erosion/Blister Scores (p < 0.01); and the levels of NLR and PLR were both positively correlated to BPDAI without Damage Score (both p < 0.05) and BPDAI Total Score (both p < 0.05). No correlation was found in other statistical analyses between hematological inflammatory markers and clinical characteristics in BP patients involved in the present study. Therefore, NLR and PLR are positively correlated with disease activity of BP.

The correlation between IgE anti-BP180 NC16A autoantibody and disease activity of bullous pemphigoid (BP) remains disputable. To determine the levels of IgE anti-BP180 NC16A autoantibody and its clinical significance in untreated BP patients. IgG and IgE anti-BP180 NC16A autoantibody in serum and blister fluid samples of 34 untreated BP patients was detected by enzyme-linked immunosorbent assay (ELISA), and correlation with clinical and pathological features of BP were statistically analysed. The Bullous Pemphigoid Disease Area Index (BPDAI) was used to measure disease activity of BP. The mean baseline level of IgG anti-BP180 NC16A autoantibody in serum and blister fluid samples of untreated BP patients was 75.3 U/mL and 1.54 U/mL, respectively (A450, cutoff: 0.126). IgE anti-BP180 NC16A autoantibody was positive in 21.9% serum and 14.7% blister fluid samples of untreated BP patients. IgE anti-BP180 NC16A autoantibody levels in serum samples positively correlated with those from blister fluid samples (r = 0.983, p < 0.05). However, IgE anti-BP180 NC16A autoantibody level in both serum and blister fluid samples of untreated BP patients did not correlate with IgG anti-BP180 NC16A autoantibody, age, extent of elevated peripheral blood eosinophils, BPDAI erosion/blister score, BPDAI urticaria/erythema score, BPDAI pruritus score, BPDAI without damage score, or BPDAI total score (all p > 0.05). No significant correlation was identified between disease activity and positive or negative anti-BP180 NC16A IgE autoantibody. Conclusion: IgE anti-BP180 NC16A autoantibody in both serum and blister fluid samples does not appear to correlate with disease activity of BP.

Objective:To analyze clinical and immunoserological features of patients with anti-p200 pemphigoid.Methods:Clinical data were collected from patients with confirmed anti-p200 pemphigoid in Hospital of Dermatology, Chinese Academy of Medical Sciences from January 2015 to October 2021, and their clinical and immunoserological characteristics were retrospectively analyzed.Results:Seven patients with anti-p200 pemphigoid were included. Indirect immunofluorescence on salt-split skin (IIF-SSS) showed that serum IgG antibodies of the 7 patients were located in the dermis of the salt-split skin, and Western blot analysis with dermal extracts as substrates revealed a protein band with a relative molecular mass of 200 000. Four patients presented with classic bullous pemphigoid-like skin lesions, 2 initially presented with eczematous lesions, and 1 presented with linear IgA bullous dermatosis-like skin lesions. Circulating IgG antibodies could recognize the recombinant laminin γ1 C-terminal region in 6 cases. Four patients received different doses of systemic glucocorticoids, 1 of whom was resistant to high-dose systemic glucocorticoids (equivalent to 1.4 mg·kg -1·d -1 prednisone) ; 2 responded well to minocycline and dapsone; 1 was lost to follow-up. Four patients achieved complete remission and discontinued the treatment at a mean follow-up of 22.5 months; 2 received complete remissiona on minimal therapy at a mean follow-up of 8 months. Conclusion:Patients with anti-p200 pemphigoid presented with heterogeneous clinical manifestations, and the recombinant C-terminal fragment of laminin γ1 can serve as a reliable antigen substrate for the detection of autoantibodies in patients with anti-p200 pemphigoid; some patients can eventually achieve complete remission off treatment.

Objective:To optimize indirect immunofluorescence on salt-split skin (IIF-SSS), and to evaluate its performance in detection of bullous pemphigoid (BP) antibodies.Methods:Normal human foreskin and non-foreskin skin tissues were used to prepare salt-split substrates under 3 different experimental conditions: traditional group rotated at 4 ℃ for 48 - 72 hours, low-temperature immersion group soaked at 4 ℃ for 48 - 72 hours, room-temperature immersion group soaked at 25 ℃ (range: 23 - 27 ℃) for 24 hours. Serum samples were obtained from 20 patients with bullous pemphigoid (BP) in Hospital of Dermatology, Chinese Academy of Medical Sciences between August 2019 and August 2020, and subjected to IIF on the intact skin or salt-split substrates by using a multiple dilution method. Paired-sample t test was used for comparisons of means between two paired samples. Results:No dermal-epidermal separation was observed in the substrates prepared in the low-temperature immersion group at 48 - 72 hours, while dermal-epidermal separation occurred in the lower lamina lucida of the foreskin and non-foreskin substrates in the room-temperature immersion group and the traditional group. For the 20 patients with BP, the reciprocal end-point titers ( M[ Q1, Q3]) detected with the salt-split non-foreskin skin and salt-split foreskin in the room-temperature immersion group, and with the salt-split non-foreskin skin in the traditional group were 5 120 (2 560, 17 920), 1 280 (640, 2 560), 1 280 (640, 2 560), respectively. Moreover, 19 (95%) patients with BP showed that the reciprocal end-point titers detected with the substrates in the room-temperature immersion group were 1 - 5 times those in the traditional group ( t = 8.04, P<0.001), suggesting that the performance of salt-split skin in the room-temperature immersion group was superior to that in the traditional group in the detection of BP antibodies; however, there was no significant difference in the reciprocal end-point titers of BP antibodies between the salt-split foreskin in the room-temperature immersion group and salt-split non-foreskin skin in the traditional group ( t<0.001, P>0.05). The reciprocal end-point titers in 20 BP sera detected by conventional IIF on the intact non-foreskin skin and foreskin were 320 (160, 640) and 480 (160, 1 120), respectively; the reciprocal end-point titers detected by IIF on the salt-split foreskin and non-foreskin skin in the room-temperature immersion group, as well as on the salt-split non-foreskin skin in the traditional group, were all consistent with or 1 - 7 times higher than those detected by conventional IIF ( t = 6.47, 14.83, 5.26, respectively, all P<0.001) . Conclusion:The soaking method at room temperature 25 ℃ (23 - 27 ℃) for preparing salt-split substrates has advantages of short duration and simple procedure, and the sensitivity of IIF-SSS using the substrates prepared by this method is equal or superior to the traditional salt-split method for detecting BP antibodies.

Objective:To evaluate the value of indirect immunofluorescence on salt-split skin (IIF-SSS) in the diagnosis of bullous pemphigoid (BP) .Methods:A single-center clinical retrospective study was conducted. Totally, 163 patients with newly diagnosed BP were collected from Hospital of Dermatology, Chinese Academy of Medical Sciences from January 2013 to January 2019, so were 404 controls, including 161 with pemphigus, 67 with eczema, 26 with drug eruption, 23 with erythema multiforme, 18 with prurigo nodularis, etc. Blood samples were collected before the treatment, and IIF-SSS, BP180 NC16A enzyme-linked immunosorbent assay (ELISA) and direct immunofluorescence (DIF) assay were performed to evaluate the value of IIF-SSS in the diagnosis of BP. Measurement data were compared by using t test and Mann-Whitney test, and enumeration data were compared by using chi-square test and Fisher′s exact test or McNemar test. Results:The number of cases positive for IIF-SSS, BP180 NC16A ELISA and DIF assay was 160, 153 and 127 respectively in the BP group, and 0, 18 and 26 respectively in the control group. The sensitivities of IIF-SSS, BP180 NC16A ELISA and DIF assay for the diagnosis of BP were 98.15%, 93.86% and 77.91% respectively, and their specificities were 100%, 95.54% and 93.56% respectively. There was strong consistency in the diagnosis of BP between IIF-SSS and DIF (Kappa coefficient= 0.767, P < 0.001) . Conclusion:IIF-SSS has relatively high sensitivity and specificity for the diagnosis of BP, and can serve as a routine method for diagnosing BP.

寻常型天疱疮( pemphigus vulgaris, PV)是以全身皮肤黏膜糜烂、松弛性水疱为临床表现的慢性易复发性自身免疫性皮肤病[1].目前主要通过抑制免疫系统,减少相关性特异性抗体的产生来治疗 PV 患者,临床常用糖皮质激素、环磷酰胺、硫唑嘌呤、静注人免疫球蛋白等治疗[2].但是长期的免疫抑制会增加发生感染的风险和继发性肿瘤等严重的并发症[3].联合使用利妥昔单抗(美罗华),既可减少糖皮质激素与其他免疫抑制剂的用量,又可避免长期使用糖皮质激素及免疫抑制剂带来的严重并发症,并可在大多数患者中得到持续的缓解[4].

A 55-year-old male patient presented with tense bullae on the extremities and trunk.Histological examination revealed subepidermal vesicles and superficial dermal infiltration of eosinophils and lymphocytes.The patient was primarily diagnosed with bullous pemphigoid.However,serum autoantibodies of the patient bound to the dermal side of salt-split skin,and no serum antibodies against BP180,BP230 or type Ⅶ collagen were detected by enzyme-linked immunosorbent assay.Hence,the diagnoses of bullous pemphigoid and epidermolysis bullosa acquisita were excluded.As Western blot and immunoprecipitation analysis showed,there existed antibodies capable of binding to a dermal antigen with a relative molecular mass of 200 000 in the serum of the patient.Based on the above findings,the patient was diagnosed as anti-laminin γ1 (p200) pemphigoid.

Objective:To investigate the correlation of related antibody titers in serum of patients with pemphigus vulgaris (PV) with disease severity and activity.Methods:A total of 24 patients with active PV were collected, who firstly visited Hospital for Skin Diseases, Chinese Academy of Medical Sciences from 2012 to 2015. Pemphigus disease area index (PDAI) was evaluated in the patients with PV at active and stable stages, and serum samples were collected. Enzyme-linked immunosorbent assay (ELISA) was conducted to determine titers of pathogenic anti-desmoglein (Dsg) conformational epitope antibodies, total anti-Dsg antibodies and anti-acetylcholine receptor (AChR) antibody in serum samples. Measurement data were compared by using t test, enumeration data were compared by using Fisher′s exact test, and correlations were analyzed by using Pearson correlation analysis. Results:Among the patients with active PV, there was no significant difference between the anti-Dsg1 antibody titers (611.4 ± 136.8) and anti-Dsg1 conformational epitope antibody titers (585.5 ± 134.7, t = 0.13, P = 0.89) , but the anti-Dsg3 antibody titers (708.6 ± 130.7) were significantly higher than the anti-Dsg3 conformational epitope antibody titers (297.2 ± 54.4, t = 2.90, P < 0.01) . In addition, both the anti-Dsg1 antibody titers and anti-Dsg1 conformational epitope antibody titers were positively correlated with PDAI scores in the patients with active PV (both r = 0.54, P < 0.01) ; PDAI scores were not correlated with the anti-Dsg3 antibody titers ( r = 0.11, P = 0.62) , but positively correlated with the anti-Dsg3 conformational epitope antibody titers ( r = 0.53, P < 0.01) . Among the 20 patients with stable PV, the serum titers of anti-Dsg1 antibodies and anti-Dsg1 conformational epitope antibodies significantly decreased compared with those at their first visit; anti-Dsg3 antibody titers significantly decreased in only 7 patients, and 13 patients still had high titers of anti-Dsg3 antibodies, including 6 with declined anti-Dsg3 conformational epitope antibody titers, and 5 converted from anti-AChR antibody-positive to anti-AChR antibody-negative. Conclusions:Both the anti-Dsg1 antibody and anti-Dsg1 conformational epitope antibody titers can reflect the disease activity of PV. The disease activity was not consistent with anti-Dsg3 antibody titers in some patients, and the anti-Dsg3 conformational epitope antibody or anti-AChR antibody may facilitate evaluating the disease activity of PV.

Pemphigus is an autoimmune bullous disease, and although several diagnostic methods are now in use indirect immunofluorescence (IIF) is still considered an important tool for diagnosing pemphigus because of its convenience, repeatability, and reduced pain for patients. The goal of the present study was to evaluate the diagnostic value of IIF on normal human skin (NS), monkey esophagus (ME), and salt-split skin (SS) for better diagnosis of pemphigus. Clinical data of 70 patients with pemphigus and 56 control were collected. IIF on NS, ME, and SS were assessed separately by observing fluorescein deposition and comparing its differentiation to different kinds of pemphigus and its sensitivities and specificities to different substrates. Intercellular deposition of IgG was visible when IIF on NS, ME, and SS were positive in patients with pemphigus. Their corresponding sensitivities and specificities were 30.0%, 84.3%, and 70.0% and 96.4%, 96.4%, and 94.6%, respectively. The differences in sensitivity were statistically significant between NS and ME and between NS and SS (P<0.001) and the specificities among the three substrates were not statistically significantly different (P>0.05). As for different types of pemphigus, the sensitivities between NS and ME and between NS and SS were statistically significantly different in both Dsg1- and Dsg3-positive and only Dsg1-positive patients with pemphigus (P<0.01); the sensitivities between NS and ME were statistically significantly different only in Dsg3-positive patients with pemphigus (P<0.001); there were no statistically significant differences between ME and SS. We therefore propose that ME is a good substrate for pemphigus diagnosis with higher sensitivity and superior to NS, particularly for patients with anti-Dsg3 antibodies. SS is a good alternative substrate to ME with almost identical higher sensitivities and specificities for diagnosis of pemphigus.

Skin diseases manifesting as desquamative gingivitis (DG) can be divided into recurrent DG-and chronic DG-related skin diseases,including oral lichen planus,mucosal pemphigoid,pemphigus vulgaris and so on.A thorough medical history,detailed oral and histopathological examinations and serum immunological tests can be helpful for correct diagnosis of DG-related skin diseases.The treatment of DG-related skin diseases includes topical and systemic therapies.It is necessary to individualize treatment protocols due to treatment response.During the treatment of DG,oral hygiene should be strengthened,secondary fungal and bacterial infections should be avoided,and attention should be paid to the protection of oral cavity and periodontal tissues.

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.

Objective To evaluate the value of indirect immunofluorescence (IIF) on three different substrates including normal human skin (NS),monkey esophagus (ME) and salt-split human skin (SS) in the diagnosis of autoimmune subepidermal bullous diseases.Methods A total of 56 patients with autoimmune subepidermal bullous diseases,including 47 with bullous pemphigoid (BP),6 with epidermolysis bullosa acquisita (EBA),2 with linear IgA bullous dermatosis,and 1 with anti-P200 pemphigoid,were diagnosed in and enrolled from Department of Dermatology,Institute of Dermatology,Chinese Academy of Medical Sciences between January 2015 and December 2016.Seventy patients with pemphigus,15 patients with chronic eczema and 15 healthy adults served as controls.Blood samples collected from these patients and controls were subjected to IIF on three different substrates including NS,ME and SS,and the fluorescence deposition was observed.The sensitivities and specificities of IIF in the diagnosis of different subepidermal bullous diseases were compared.Statistical analysis was carried out with SPSS 13.0 software by using chi-square test for the comparison of enumeration data.Results IIF on NS or ME in the serum of patients with BP showed linear deposition of fluorescent material along the basement membrane zone.IIF on SS showed linear deposition of fluorescent material in the epidermis in the patients with BP,but in the dermis in the patients with EBA and anti-P200 pemphigoid.The sensitivities of IIF on NS,ME or SS in the diagnosis of subepidermal bullous diseases were 73.2％,60.7％ and 94.6％ respectively,and the specificities were 98.0％,100％ and 97.1％ respectively.There were significant differences among the sensitivities (x2 =18.2,P ＜ 0.05),but no significant difference was observed among the specificities (P ＞ 0.05).The diagnostic sensitivity of IIF on SS was significantly higher than that of IIF on NS or ME(x2 =8.0,16.7,both P ＜ 0.05).Conclusion In the diagnosis of autoimmune subepidermal bullous diseases,IIF on SS is superior to IIF on ME or NS.

Objective To prepare human epidermal extracts by thermal separation,and to evaluate the value of epidermal extract-based Western blot analysis in the diagnosis of bullous pemphigoid (BP).Methods Human epidermal extracts were prepared by thermal separation from circumcised foreskins of healthy males.Serum samples were obtained from 22 inpatients with BP and 25 inpatients without BP in Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College between January 2015 and August 2017.These serum samples were subjected to Western blot analysis with epidermal extracts as substrates,as well as to BP180-NC16A enzyme-linked immunosorbent assay (ELISA).Statistical analysis was carried out using chi-square test and Fisher's exact test with the SPSS22.0 software.Results The sensitivities of epidermal extract-based Western blot analysis and BP 180-NC16A ELISA in the diagnosis of BP were 86.36％ (95 ％ CI:64.03％-96.41％) and 95.45％ (95％ CI:75.11％-99.76％) respectively (~ =1.10,P =0.294),and the specificities were 100％ (95％ CI:83.42％-100％) and 92％ (95％ CI:75.11％-99.76％) respectively (x2 =20.8,P =0.149).Epidermal extract-based Western blot analysis in the 22 patients with BP showed a protein band with relative molecular mass (RMM) of 230 000 in 4 patients,a protein band with RMM of 180 000 in 18,a protein band with RMM of 120 000 in 1,and a protein band with RMM of 97 000 in 1.The BP180-NC16A ELISA showed that the antibody titers were more than 50 U/ml in the BP patients with protein bands of RMM of 180 000.Conclusions The epidermal extract-based Western blot analysis mainly showed the protein band with RMM of 180 000 in the patients with BP.The sensitivity of the epidermal extract-based Western blot analysis was lower than that of the BP180-NC16A ELISA,and the epidermal extract-based Western blot analysis tends to be negative when the titer of the autoantibody is low.

大疱性类天疱疮(bullous pemphigoid,BP)是老年人群当中最常见的自身免疫性水疱病,它的主要特点是血清中存在抗基底膜带抗体,与基底膜带相应抗原结合,通过炎症细胞的激活破坏基底膜带,从而引起病理或肉眼可见的水疱,可累及皮肤和粘膜.BP180是BP的最重要的抗原.随年龄增长,BP发病率逐渐提高.老年人发生BP后,有较高的死亡率.BP临床表现主要是紧张性的瘙痒性水疱,部分患者临床表现不典型,可不出现明显水疱.BP的诊断依赖于临床表现、病理表现、免疫病理表现和血清学检查.BP在治疗上,目前首选的治疗是局部或者系统用糖皮质激素,部分患者对非激素治疗也可能有效.

Immunoglobulin A (IgA) is the most common subtype of antibodies in mucosal surfaces. In most of autoimmune bullous diseases, however, immunoglobulin G (IgG) is the main pathogenic antibody that plays a role through complementation. The IgA antibody for epidermal connection protein can be found in the sera of some patients with blistering skin disease. Of these patients, some have the IgA antibody in their sera, while others have IgG and IgA antibodies. IgA-related autoimmune bullous diseases are less common in clinical practice. In the past, these diseases were not fully understood and their classifications were confusing. Recently, some progress has been made in the study of these diseases.

Objective To evaluate the value of indirect immunofluorescence on salt-split skin (IIF-SSS) and bullous pemphigoid 180 N C 16a enzyme-linked immunosorbent assay (BP 180 N C 16a-ELISA) in the diagnosis of bullous pemphigoid (BP).Methods Serum samples were collected from 174 BP patients and 129 controls,who were enrolled from Institute of Dermatology of Chinese Academy of Medical Sciences and Peking Union Medical College between January 2015 and August 2017,and subjected to IIF-SSS and BP180 NC16a-ELISA.Direct immunofluorescence (DIF) test was performed in 25 cases of BP,and its sensitivity for the diagnosis of BP was compared with that of IIF-SSS and BP180 NC16a-ELISA.Results The sensitivities for IIF-SSS and BP180 NC16a-ELISA were 93.67％ and 96.55％ respectively,and the specificities for IIF-SSS and BP180 NC16a-ELISA were 100％ and 96.12％ respectively.IIF-SSS was weakly correlated with BP180 NC16a-ELISA with a correlation coefficient of 0.147.There was no significant difference in the sensitivity between the serological diagnostic methods (IIF-SSS and BP180 NC 16a-ELISA) and DIF.Conclusion Serological diagnostic methods show high specificity and sensitivity in the diagnosis of BP,and are worthy of clinical promotion and application.