Hu W Y, Huang Y Y, Liu T, Cai Y
Department of Periodontal Mucosa, Guizhou Medical University School of Stomatology, Guiyang 550004, China.
Department of Orthodontics, West China Hospital of Stomatology, Sichuan University & State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, Chengdu 610041, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2020 Feb 9;55(2):80-85. doi: 10.3760/cma.j.issn.1002-0098.2020.02.002.
To investigate the effects of exogenous interleukin (IL)-35 on the balance of helper T cell 17 (Th17) and regulatory T cell (Treg) in peripheral blood of patients with oral lichen planus (OLP). Totally 12 peripheral blood samples of OLP patients (OLP group, one male and 11 female, 26-68 years old; four cases of reticular OLP and eight cases of erosive OLP) were collected from patients of Department of Oral Mucosal Specialist of the Affiliated Hospital of Guizhou Medical University from October to December 2016. During the same period, thirteen normal peripheral blood samples were collected from the Physical Examination Center of the Affiliated Hospital of Guizhou Medical University (normal control group, one male and 12 female, 20-68 years old). The peripheral blood mononuclear cells (PBMC) were extracted in sterile condition and CD4+ T cells were sorted by flow cytometry (FCM). Quantitative real-time PCR (qPCR) technique was used to detect the mRNA expression levels of retinoid-related orphan nuclear γt (RORγt) and forkhead box3 (Foxp3). The CD4+ T cells were divided into experimental group and control group. The CD4+ T cells of experimental group were cultured by adding rhIL-35, and the CD4+ T cells of control group were cultured with the same volume of phosphate buffered saline (PBS). After the completion of the culture, the cells were collected. The expression levels of the same factors were detected by qPCR. The expression [((25), (75))] of Foxp3 [0.15 (0.09, 0.30)] and RORγt mRNA [1.04 (0.45, 2.15)] in the CD4+ T cells of OLP were significantly higher than those in normal control group [0.04 (0.02, 0.06), 0.10 (0.05, 0.11)] (-4.134, <0.01; 3.699, <0.01). The ratio of ROR γt/Foxp3 mRNA in OLP group [6.22(3.67, 15.34)] was higher than that in normal control group [2.50 (1.24, 5.23)] (-2.665, 0.007). In the CD4+ T cells of OLP patients, the expression of Foxp3 mRNA in the experiment group [0.40 (0.21, 1.22)] was higher than that in the control group [0.15 (0.11, 0.26)](-2.510, 0.012), and the expression of ROR γt mRNA between two groups showed no significant difference (>0.05). The ROR γt/Foxp3 mRNA ratio [3.44 (1.55, 8.16)] of the experiment group was lower than that in the control group [6.22 (4.43, 12.21)] (-2.746, 0.006). There was a Th17/Treg imbalance with predominated by Th17 cells in the peripheral blood of patients with OLP. Exogenous rhIL-35 had an immunomodulatory effect on the balance of Th17/Treg.
探讨外源性白细胞介素（IL）-35对口腔扁平苔藓（OLP）患者外周血中辅助性T细胞17（Th17）和调节性T细胞（Treg）平衡的影响。于2016年10月至12月，从贵州医科大学附属医院口腔黏膜科患者中收集12例OLP患者的外周血样本（OLP组，男1例，女11例，年龄26 - 68岁；网状OLP 4例，糜烂型OLP 8例）。同期，从贵州医科大学附属医院体检中心收集13例正常外周血样本（正常对照组，男1例，女12例，年龄20 - 68岁）。在无菌条件下提取外周血单个核细胞（PBMC），并通过流式细胞术（FCM）分选CD4⁺T细胞。采用定量实时聚合酶链反应（qPCR）技术检测维甲酸相关孤儿核γt（RORγt）和叉头框3（Foxp3）的mRNA表达水平。将CD4⁺T细胞分为实验组和对照组。实验组CD4⁺T细胞加入重组人IL-35（rhIL-35）进行培养，对照组CD4⁺T细胞用等体积的磷酸盐缓冲液（PBS）培养。培养结束后收集细胞，用qPCR检测相同因子的表达水平。OLP患者CD4⁺T细胞中Foxp3[0.15（0.09，0.30）]和RORγt mRNA[1.04（0.45，2.15）]的表达[（（25），（75））]显著高于正常对照组[0.04（0.02，0.06），0.10（0.05，0.11）]（-4.134，<0.01；3.699，<0.01）。OLP组RORγt/Foxp3 mRNA比值[6.22（3.67，15.34）]高于正常对照组[2.50（1.24，5.23）]（-2.665，0.007）。在OLP患者CD4⁺T细胞中，实验组Foxp3 mRNA表达[0.40（0.21，1.22）]高于对照组[0.15（0.11，0.26）]（-2.510，0.012），两组RORγt mRNA表达差异无统计学意义（>0.05）。实验组RORγt/Foxp3 mRNA比值[3.44（1.55，8.16）]低于对照组[6.22（4.43，12.21）]（-2.746，0.006）。OLP患者外周血中存在以Th17细胞为主导的Th17/Treg失衡。外源性rhIL-35对Th17/Treg平衡具有免疫调节作用。
