上海交通大学学报（医学版）

上海交通大学学报(医学版) ›› 2020, Vol. 40 ›› Issue (05): 626-632.doi: 10.3969/j.issn.1674-8115.2020.05.010

• 论著·临床研究 • 上一篇    下一篇

舌下含服免疫治疗对效应性T细胞和调节性T细胞功能的影响

戴 飞，魏瑾瑾，汤欣玥，陈 峥，蔺 林   

  1. 复旦大学附属华山医院北院耳鼻咽喉头颈外科，上海201907
  • 出版日期:2020-05-28 发布日期:2020-05-28
  • 通讯作者: 蔺 林，电子信箱：linlinhsn@aliyun.com。
  • 作者简介:戴 飞（1987—），男，主治医师，硕士；电子信箱：hsdaifei@aliyun.com。
  • 基金资助:
    国家自然科学基金（81371076）；上海申康医院发展中心郊区医院临床能力建设项目（SHDC12015905）。

Effect of sublingual immunotherapy on functions of effector T cells and regulatory T cells

DAI Fei, WEI Jin-jin, TANG Xin-yue, CHEN Zheng, LIN Lin   

  1. Department of Otorhinolaryngology-Head and Neck Surgery, Huashan Hospital North of Fudan University, Shanghai 201907, China
  • Online:2020-05-28 Published:2020-05-28
  • Supported by:
    National Natural Science Foundation of China (81371076); Shanghai Suburb Tertiary Hospital Clinical Capacity Building Project (SHDC12015905).

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摘要： 目的·研究舌下含服免疫治疗（sublingual immunotherapy，SLIT）对变应性鼻炎（allergic rhinitis，AR）炎症反应中效应性T细胞（effector T cell，Teff）和调节性T细胞（regulatory T cell，Treg）功能的影响。方法·选择复旦大学附属华山医院北院耳鼻咽喉头颈外科招募的20例AR患者，分别在治疗前及治疗24个月和30个月时获取其外周血单个核细胞（peripheral blood mononuclear cell，PBMC），检测PBMC中Teff和Treg所占CD4+ T细胞百分比，并检测PBMC中2种细胞钙释放激活钙通道调节分子1（calcium release-activated calcium channel modulator 1，ORAI1）蛋白的浓度和Ca2+平均荧光强度（mean fluorescence intensity，MFI）。然后将治疗30个月时的2种细胞进行体外培养，检测2种细胞中ORAI1蛋白的表达以及培养基中白细胞介素-4（interleukin-4，IL-4）和IL-10浓度。制备含人ORAI1短发夹RNA的慢病毒并将其转染至2种细胞，再次检测2种细胞中Ca2+ MFI和培养基中IL-4和IL-10的浓度。结果·在治疗24个月和30个月时PBMC中Teff所占CD4+ T细胞百分比逐渐减少（P=0.000，P=0.027），而Treg所占CD4+ T细胞百分比逐渐增多（P=0.000，P=0.008）；Teff中ORAI1蛋白表达量（P=0.000，P=0.003）和Ca2+ MFI（P=0.000，P=0.000）也逐渐减少，而Treg中ORAI1蛋白表达量（P=0.000，P=0.007）和Ca2+ MFI（P=0.000，P=0.000）随着治疗时间延长逐渐增多。体外培养的2种细胞均表达ORAI1蛋白，而且在慢病毒转染后，细胞中Ca2+ MFI降低（P=0.004，P=0.000），培养基中IL-4（P=0.009）和IL-10（P=0.000）的浓度均显著降低。结论·SLIT可以通过调节ORAI1蛋白的表达控制Teff和Treg细胞的功能。

关键词: 免疫治疗, 变应性鼻炎, T细胞, 钙释放激活钙通道调节分子1, 细胞因子

Abstract:

Objective · To investigate the effect of sublingual immunotherapy (SLIT) on the functions of effector T cells (Teff) and regulatory T cells (Treg) under allergic rhinitis (AR) condition. Methods · Peripheral blood mononuclear cells (PBMC) were obtained from 20 AR patients who were recruited from Department of Otorhinolaryngology-Head and Neck Surgery, Huashan Hospital North of Fudan University before and after the 24- and 30-month treatments of SLIT. The percentages of Teff and Treg in CD4+ T cells in PBMC were examined and the contents of calcium release-activated calcium channel modulator 1 (ORAI1) protein and Ca2+ mean fluorescence intensity (MFI) in PBMC were measured. Then, Teff and Treg after the 30-month treatment were cultured in vitro, and were evaluated for the expression of ORAI1 protein and concentrations of interleukin-4 (IL-4) and IL-10 in the cultures. Lentivirus vector (lenti) that encoded short hairpin RNA against ORAI1 (lenti-ORAI1) was prepared and transfected into Teff and Treg, and then Ca2+ MFI in Teff and Treg and concentrations of IL-4 and IL-10 in the cultures were assessed again. Results · The percentage of Teff in CD4+ T cells in PBMC reduced gradually after the 24- and 30-month treatments (P=0.000, P=0.027); however, the percentage of Treg was increased (P=0.000, P=0.008). The expression of ORAI1 protein (P=0.000, P=0.003) and Ca2+ MFI (P=0.000, P=0.000) in Teff were also decreased gradually after the 24- and 30-month treatments; however, the expression of ORAI1 protein (P=0.000, P=0.007) and Ca2+ MFI in Treg were enhanced (P=0.000, P=0.000). ORAI1 protein was expressed in these two types of cells cultured in vitro. Furthermore, Ca2+ MFI in them (P=0.004, P=0.000) and IL-4 (P=0.009) and IL-10 (P=0.000) in the cultures were all decreased after the transfection of lenti-ORAI1. Conclusion · SLIT can control functions of Teff and Treg through regulating the expression of ORAI1 protein.

Key words: immunotherapy, allergic rhinitis, T cell, calcium release-activated calcium channel modulator 1 (ORAI1), cytokine

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