中华皮肤科杂志 ›› 2018, Vol. 51 ›› Issue (4): 309-312.doi: 10.3760/cma.j.issn.0412-4030.2018.04.015

• 研究报道 • 上一篇    下一篇

天疱疮患者皮损局部B淋巴细胞及其产生特异性抗体的功能研究

元慧杰1,周生儒1,刘芝翠1,朱海琴2,许人超3,郑捷1,潘萌1   

  1. 1. 上海交通大学医学院附属瑞金医院皮肤科
    2. 上海交通大学医学院附属瑞金医院
    3. 上海市嘉定区中心医院
  • 收稿日期:2017-03-16 修回日期:2017-11-05 发布日期:2018-03-29
  • 通讯作者: 潘萌 E-mail:panmeng2000@yahoo.com
  • 基金资助:
    国家自然科学基金;国家自然科学基金;国家自然科学基金;上海市科研计划项目;上海交通大学医学院-高峰高原计划-“研究型医师”项目

Expression and specific antibody-producing capacity of B lymphocytes infiltrating in lesions of patients with pemphigus vulgaris

  • Received:2017-03-16 Revised:2017-11-05 Published:2018-03-29
  • Supported by:
    National Natural Science Foundation of China;National Natural Science Foundation of China;National Natural Science Foundation of China;Plan of Shanghai Municipal Science and Technology Commission;Shanghai Jiao Tong University School of Medicine-Gaofeng Clinical Medicine Grant Support

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摘要： 目的 检测寻常型天疱疮患者皮损局部B淋巴细胞及其产生特异性抗体的功能。方法 寻常型天疱疮患者35例，健康对照22例。取健康对照皮肤和寻常型天疱疮患者初发水疱或糜烂皮损组织，分离获得单个核细胞。流式细胞仪检测患者皮损局部淋巴细胞、CD19+ B细胞比例，以及特异性识别桥粒芯糖蛋白1（Dsg1）和Dsg3的CD19+ B淋巴细胞比例。体外培养寻常型天疱疮患者皮损局部淋巴细胞，ELISA法检测培养上清液中抗Dsg1和Dsg3抗体滴度，采用受试者工作特征曲线（ROC）分析阳性率。结果 寻常型天疱疮患者皮损局部淋巴细胞、CD19+ B细胞比例分别为17.95% ± 3.85%、4.27% ± 1.13%，高于健康对照组（7.83% ± 1.29%、0.61% ± 0.31%），差异有统计学意义（t = 2.49，U = 13.00，均P < 0.05）。天疱疮患者皮损局部CD19+ B淋巴细胞中，表达IgG的细胞比例为（38.33 ± 5.56）%，表面识别Dsg1与Dsg3的细胞比例分别为12.87% ± 1.267%、10.42% ± 1.243%。局部淋巴细胞体外培养6 d后，培养上清液中抗Dsg1与Dsg3抗体滴度分别为（4.89 ± 1.56） U/ml、（35.45 ± 13.03） U/ml，阳性率分别为85%（17/20）和95%（19/20）。 结论 寻常型天疱疮患者皮损局部有可与Dsg1及Dsg3特异性结合的B淋巴细胞聚集，体外培养后可产生特异性抗Dsg1与抗Dsg3自身抗体。

关键词: 天疱疮, B淋巴细胞, 桥粒芯糖蛋白质1, 桥粒芯糖蛋白质3, 免疫球蛋白G

Abstract: Yuan Huijie, Zhou Shengru, Liu Zhicui, Zhu Haiqin, Xu Renchao, Zheng Jie, Pan Meng Department of Dermatology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China Corresponding author: Pan Meng, Email: panmeng@medmail.com.cn 【Abstract】 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, CD19+ 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 CD19+IgG+ 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 supernatant 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.

Key words: Pemphigus, B-lymphocytes, Desmoglein 1, Desmoglein 3, Immunoglobulin G

中图分类号: 

  • R758.66

引用本文

元慧杰 周生儒 刘芝翠 朱海琴 许人超 郑捷 潘萌. 天疱疮患者皮损局部B淋巴细胞及其产生特异性抗体的功能研究[J]. 中华皮肤科杂志, 2018,51(4):309-312. doi:10.3760/cma.j.issn.0412-4030.2018.04.015