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Objective:To investigate the local infiltration of tissue-resident memory CD4 + T (CD4 + T RM) cells in lesions of patients with pemphigus and its clinical implications. Methods:From September 2017 to December 2018, 20 patients with pemphigus and 15 healthy human controls were collected from Department of Dermatology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine. Flow cytometry was performed to determine the proportion of CD4 + T RM cells in skin lesions of pemphigus patients and normal skin of healthy controls. The degree of CD4 + T RM cell infiltration in skin lesions was compared among different body sites of the patients with pemphigus, and the correlation between the proportion of CD4 + T RM cells and the time to disease control was analyzed. Normally distributed data were analyzed by using t test, and non-normally distributed data by using non-parametric test; the Pearson correlation coefficient was used to analyze correlations of the proportion of CD4 + T RM cells with pemphigus disease area index (PDAI) scores and circulating anti-desmoglein (Dsg) antibody titers. Results:Among the 20 patients, there were 16 with pemphigus vulgaris and 4 with pemphigus foliaceus. All the patients had skin involvement, 14 lesional tissue samples were taken from the trunk, and 6 from the limbs. There was no significant difference between the healthy control group and pemphigus group in terms of age, gender or biopsy sites (all P > 0.05) . The proportions of CD3 + T cells (72.75% ± 8.22%) and CD4 + T RM cells (44.05% ± 14.27%) in the skin lesions of patients with pemphigus were significantly higher than those in the skin tissues of the healthy controls (31.33% ± 8.72%, 12.60% ± 5.12%, t = 14.24, 9.10, respectively, both P < 0.001) . Among the patients with pemphigus, the proportion of CD4 + T RM cells was significantly higher in the skin lesions on the trunk (49.57% ± 12.32%) than in those on the limbs (31.17% ± 9.75%, t = 3.23, P < 0.05) . The proportion of CD4 + T RM cells in the skin lesions was positively correlated with the PDAI scores ( r2 = 0.246, P = 0.026) , but not correlated with serum titers of circulating anti-Dsg1 ( r2 = 0.137, P > 0.05) or anti-Dsg3 ( r2 = 0.162, P > 0.05) antibodies in the patients. During the treatment with systemic glucocorticoids, the proportion of CD4 + T RM cells in the skin lesions was significantly higher in the patients whose lesions could not be controlled within 4 weeks than in those whose lesions could be controlled within 4 weeks ( t = 3.22, P < 0.05) . Conclusion:The proportion of CD4 + T RM cells markedly increased in the skin lesions of patients with pemphigus, which may be related to the severity of the disease and response to treatment.
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Objective To assess the association between early adiposity rebound (AR) and indices of obesity and metabolic risk in 5-year-old children. Methods Based on Ma’anshan Birth Cohort Study (MABC), single live births born in Ma'anshan of Anhui province from October 2013 to April 2015 were followed for up to 5 years consecutively. As of August 2019, 720 children with continuous measurements (≥8 times) and metabolic indicators were obtained. Physical examination and laboratory tests were used to obtain information on the birth status, length/height, weight, waist circumference, body composition and metabolic indicators of children. The 2 test, F test, t-test, non-parametric test, general linear model and logistic regression model were used for statistical analysis. Results 43.5% of the children had AR≤4 years. After controlling for gender, it was found that earlier AR was associated with overweight/obesity (OR=2.71, 95%CI: 1.81~4.05), larger waist circumference (OR=1.88, 95%CI: 1.25~2.82), and body fat percentage ≥90th percentile (OR=2.09, 95%CI: 1.26~3.48). In the earlier AR group, the insulin resistance and metabolic score were higher, but the difference was not statistically significant. At 5 years of age, the prevalence of obesity and overweight was 6.0% and 12.8%, respectively. Children with overweight/obesity, larger waist circumference, higher waist-to-weight ratio and body fat percentage ≥ 90th percentile were associated with higher insulin resistance and metabolic score, and all the differences were statistically significant (all P<0.001). Conclusion Earlier AR increased the risk of overweight/obesity, larger waist circumference, and body fat percentage ≥90th percentile at age of 5 years. Each index of the commonly used measures of childhood obesity was closely related with insulin resistance and metabolic risk factors at 5 years old.
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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.
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Objective To summarize clinical and histopathological features of trauma-triggered autoimmune bullous skin diseases,and to explore its possible pathogenesis.Methods Clinical manifestations,histopathological features and treatment of 3 cases of trauma-triggered pemphigus or pemphigoid were analyzed,and summarized according to related domestic and overseas literature.Results Of the 3 cases,1 was a female aged 62 years,and 2 were males aged 60 and 71 years respectively.They all had a history of skin trauma or surgery before the onset of the diseases,and time intervals from trauma to diseases were 5,5 weeks and 3 days respectively.The 3 cases were diagnosed as bullous pemphigoid (anti -BP180 antibody 109 U/ml,anti-BP230 antibody negative),pemphigus vulgaris (anti-Dsg1 antibody 68.8 U/ml,anti-Dsg3 antibody 219 U/ml) and pemphigus foliaceus (anti-Dsg1 antibody 143 U/ml,anti-Dsg3 antibody negative) respectively.Their lesions were relieved dramatically after oral and (or) topical glucocorticoid treatment.Conclusions Trauma may be a triggering factor for autoimmune bullous skin diseases.For patients with post-traumatic poor wound healing or skin erythema,blisters and erosion,which can hardly be attributed to trauma or surgery itself,autoimmune bullous skin diseases should be considered,and histopathological or immunopathological examinations should be performed timely.
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Objective To study the changes of antigen-specific T helper type 1 (Th1) cells and Th2 cells in patients with pemphigus vulgaris (PV) at different stages,so as to better understand the roles of autoreactive T cells in PV.Methods The DG3 (96-112) peptide was synthesized.Twenty-four patients with PV and 10 health checkup examinees were included in this study.Peripheral blood mononuclear cells (PBMCs) were obtained from the health checkup examinees and all the patients before treatment and seven patients about one month after treatment,and stimulated with the DG3 peptide of 10 mg/L for different durations.Then,enzyme-linked immunospot (ELISPOT) assay was performed to count the number of DG3-specific IFN-γ+ Th1 cells,IL-4+ Th2 cells and memory B cells.One-way analysis of variance (ANOVA) and t test were done to compare the number of cells between different groups,and Pearson correlation coefficient was used to evaluate the correlation among Th cells,memory B cells and anti-desmoglein 3 (Dsg3) antibody titers.Results In these patients,the male to female ratio was 1.67 ∶ 1,and the average age was (56.59 ± 14.66) years.Compared with the health check-up examinees,the patients with PV showed a higher absolute number of DG3-specific IFN-γ+ Th1 cells (420.18 ± 350.29 vs.145.12 ± 86.56,t =3.25,P ＜ 0.05),but a similar absolute number of specific IL-4+ Th2 cells (366.76 ± 192.44 vs.335.88 ± 164.96,P＞ 0.05) per 5 × 105 PBMCs.The percentage of DG3-specific IL-4+ Th2 cells in Th2 cells was 37.03％ ± 23.44％,and the percentage of IFN-γ+ Th1 cells was 23.62％ ± 16.77％ in peripheral blood of patients with PV.The number of DG3-specific IL-4+ Th2 cells per 5 × 105 PBMCs significantly decreased from 452.82 ± 199.29 before treatment to 241.68 ± 160.60 after treatment in seven patients (t =2.48,P ＜ 0.05).There was no significant correlation between specific Th cells,memory B cells and anti-Dsg3 antibody titers (all P ＞ 0.05).Conclusions The peptide DG3 (96-112) has pathogenic epitopes which could be recognized by specific Th cells of patients with PV.Both antigen-specific IFN-γ+ Th1 cells and IL-4+ Th2 cells play certain roles in the pathogenesis of pemphigns vulgaris,and IL-4+ Th2 cells appear to be more important.
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Objective To explore the mechanisms underlying the mediating effect of regulatory B (Breg)cells in the pathogenesis of pemphigus.Methods Peripheral blood specimens were collected from 48 patients with pemphigus at different degrees of severity and 20 healthy controls.Flow cytometry was conducted to analyze the frequency of Breg(CD19 +CD24hiCD38hi)cells in peripheral CD19+ B cells,enzyme linked immunosorbent assay(ELISA)to determine the titer and subtypes of anti-desmoglein(Dsg)1 and 3 antibodies in sera from these subjects.The relationship of anti-Dsg 1 and 3 antibodies with the frequency of Breg cells was assessed by Spearman's rank correlation test.Results Significant differences were observed in the proportion of Breg cells in CD19+ B cells between patients with pemphigus and healthy controls(11.54％ ± 0.97％ vs.8.19％ ± 0.85％,P =0.04),as well as between patients with acute mild pemphigus and those with moderate and severe pemphigus(5.17％ ± 2.14％ vs.11.38％ ± 5.30％ and 11.17％ ± 5.31％,P =0.042,0.046,respectively).The frequency of Breg cells was positively correlated with the absorbance value for anti-Dsg1 antibodies of IgG4 subclass and IgG4/IgG1 ratio(r =0.527,0.565,respectively,both P ＜ 0.01).Conclusions Breg cells are mainly associated with the production of antibodies in and disease severity of pemphigus,while the actual mechanism of action remains unknown.
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Objective To assess the correlation between the subclasses of antibodies against desmoglein (Dsg) 1 and Dsg3 and disease activity in patients with pemphigus. Methods Sera were collected from 47 patients with pemphigus, and ELISA was performed to determine the titers and subclasses of antibodies against Dsg1 and Dsg3. The correlation of antibody titers and subclasses with disease activity was assessed.Results Clinical phenotype was associated with antibody profiles in these patients with pemphigus. Of 17 patients with mucocutaneous involvement, 14 (82.4%) had both anti-Dsg3 and anti-Dsg1 antibodies; of 16 patients with cutaneous involvement, 15 (93.7%) had anti-Dsg1 antibody, only 1 (6.3%) developed anti-Dsg3 antibody; of 6 patients with mucosal involvement, all ( 100% ) had only anti-Dsg3 antibody. The serum levels of antibodies against Dsg1 and Dsg3 were increased, but not in parallel with the disease severity in these patients.Moreover, the subclasses of antibodies were correlated with disease severity. IgG4 subclass antibodies against Dsg1 and Dsg3 predominated in patients with pemphigus at active stage, whereas IgG1 subclass in those at remission stage. The serum levels (expressed as absorbance value) of IgG4 and IgG1 subclass antibodies were 1.92 ± 1.21 and 0.60 ± 0.61 respectively, with the ratio of IgG4 to IgG1 more than 1, in patients with antiDsg1 antibodies at active stage, 0.03 ± 0.02 and 0.22 ± 0.11 respectively with the ratio of IgG4 to IgG1 less than 1, in those at remission stage, 2.35 ± 2.17 and 1.84 ± 1.16 respectively with the ratio of IgG4 to IgG1 more than 1 in patients with anti-Dsg3 antibodies at active stage, 0.15 ± 0.16 and 1.05 ± 0.77 respectively with the ratio of IgG4 to IgG1 less than 1 in those at remission stage. Conclusions The subclasses of pemphigus-specific antibodies are closely correlated with the disease severity in patients with pemphigus. The detection of subclasses and titers of anti-Dsg1 and anti-Dsg3 antibodies may aid in the diagnosis of and monitoring of disease severity in pemphigus.
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Objective To study the efficacy of rituximab in the treatment of pemphigus vulgaris (PV) and its underlying mechanism. Methods A 38-year-old female with PV presented with refractory, painful oral ulcers and erosions. Since she was poorly responsive to prednisone 80 mg daily, intravenous ritu-ximab of 500 mg once a week was given for successive 3 weeks followed by 5 successive days of intra-venous gamma globulin at a dose of 400 mg/kg per day, and a total of two treatment sessions were conducted. ELISA was used to detect the serum titer of anti-Dsg3 antibodies and their IgG subtypes (IgG1 and IgG4) as well as serum level of interferon-gamma (IFN-gamma), interleukin-4 (IL-4), interleukin-10 (IL-10). Results Three months after the end of two treatment sessions, the symptom was obviously improved, and lesions subsided; alter another 1 month, clinical symptom fully disappeared. During the 1-year follow-up, no lesions recurred. The anti-Dsg3 antibody titer was 253.33 U/mL before treatment, plateaued at 250 U/mL within 4 weeks after the initial infusion, decreased till 3 months after the withdrawal, and reached 26.06 U/mL 7 months after the withdrawal, and remained within normal range till the time of this writing. The serum titer of IgGl subclass of anti-Dsg3 antibodies dropped dramatically fight after the first infusion, but that of IgG4 subclass remained at a high level at early stage of medication, began to decline until 3 months after the with-drawal, and finally reached the normal range following clinical remission. Also, serum level of IFN-garnma and IL-10 correlated with the severity of PV. Conclusions Combined rituximab is effective for the control of PV, likely by eliminating Dsg3-specific antibodies, especially IgG4 subclass of them.