天疱疮患者皮损中CD4^+定居记忆T细胞的浸润及其临床相关性分析

Local infiltration of tissue-resident memory CD4^+T cells in lesions of patients with pemphigus and its clinical implications

目的探讨天疱疮皮损局部CD4+定居记忆T(TRM)细胞的浸润及其临床相关性。方法2017年9月至2018年12月在上海交通大学医学院附属瑞金医院皮肤科收集天疱疮患者20例,健康对照15例。采用流式细胞仪检测天疱疮患者皮损和健康对照皮肤CD4^+TRM细胞的表达。比较天疱疮患者不同部位皮损处CD4+TRM细胞的浸润情况,分析CD4^+TRM细胞比例与病情控制所需时间的关系。符合正态分布的资料采用t检验,不符合正态分布者采用非参数检验;采用Pearson相关系数分析CD4^+TRM细胞比例和天疱疮疾病面积指数(PDAI)评分、循环中抗桥粒芯糖蛋白(Dsg)抗体滴度的相关性。结果20例天疱疮患者中16例为寻常型天疱疮,4例为落叶型天疱疮。所有患者均有皮肤累及,14例皮损组织取材于躯干部位,6例取材于四肢部位。健康对照组年龄、性别和取材部位与天疱疮组差异无统计学意义(均P>0.05)。天疱疮患者皮损局部CD3^+T细胞(72.75%±8.22%)显著高于健康对照组(31.33%±8.72%,t=14.24,P<0.001),CD4^+TRM细胞比例(44.05%±14.27%)亦显著高于健康对照组(12.60%±5.12%,t=9.10,P<0.001)。天疱疮患者躯干部CD4+TRM细胞比例(49.57%±12.32%)显著高于四肢(31.17%±9.75%,t=3.23,P<0.05)。皮损局部CD4+TRM细胞比例与PDAI呈正相关(r2=0.246,P=0.026);而与血清中循环抗Dsg1(r^2=0.137,P>0.05)和抗Dsg3(r2=0.162,P>0.05)抗体滴度无显著相关性。系统糖皮质激素治疗过程中4周内不能控制的患者皮损处CD4^+TRM细胞比例显著高于4周内可以控制的患者(t=3.22,P<0.05)。结论天疱疮患者皮损局部CD4+TRM细胞比例显著增高,可能与病情严重程度和疾病治疗反应相关。

Objective To investigate the local infiltration of tissue-resident memory CD4^+T(CD4^+TRM)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+TRM cells in skin lesions of pemphigus patients and normal skin of healthy controls.The degree of CD4^+TRM cell infiltration in skin lesions was compared among different body sites of the patients with pemphigus,and the correlation between the proportion of CD4^+TRM 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^+TRM 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^+TRM 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^+TRM 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+TRM cells in the skin lesions was positively correlated with the PDAI scores(r^2=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^+TRM 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+TRM 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.