T细胞免疫球蛋白和黏蛋白结构域蛋白3抑制银屑病患者CD14^+单核细胞分泌白细胞介素12

T cell immunoglobulin and mucin domain-containing protein 3 inhibits interleukin-12 secretion by CD14^+ monocytes from patients with psoriasis vulgaris

目的评估T细胞免疫球蛋白和黏蛋白结构域蛋白3(TIM-3)对银屑病患者外周血CD14^+单核细胞分泌白细胞介素12(IL-12)的调控。方法收集2017年11月至2018年3月在山西省人民医院就诊的寻常型银屑病患者47例(银屑病组)和健康志愿者19例(对照组),分离外周血单个核细胞,使用Toll样受体(TLR)配体刺激培养后,流式细胞仪检测CD14^+单核细胞中TIM-3的表达和IL-12的分泌情况。使用抗TIM-3中和抗体封闭TIM-3通路后观察CD14^+单核细胞下游信号通路及分泌IL-12的变化。两组间数据比较采用t检验,相关性分析采用Pearson检验。结果在未刺激条件下,CD14^+单核细胞分泌IL-12的水平较低,银屑病组CD14^+TIM-3^+细胞比例(12.20%±2.83%)显著高于对照组(9.91%±1.77%,t=3.270,P=0.0017)。TLR配体刺激后,银屑病组CD14^+IL-12^+细胞比例(13.49%±2.80%)显著低于对照组(28.97%±8.97%,t=10.71,P<0.0001),但CD14^+TIM-3^+细胞比例(6.80%±1.11%)仍显著高于对照组(4.85%±1.37%,t=6.064,P<0.0001)。对照组和银屑病组CD14^+TIM-3^+与CD14^+IL-12^+细胞比例呈显著负相关(r=-0.473和r=-0.371,P<0.05)。封闭TIM-3通路可诱导CD14^+单核细胞中细胞因子信号抑制物1表达降低,信号传导及转录激活因子1磷酸化增加,促进皮损部位分离的角质形成细胞所诱导的CD14^+单核细胞分泌IL-12增加。结论在银屑病中,TIM-3在CD14^+单核细胞介导的固有免疫应答中发挥重要的负性调控作用。

Objective To assess the role of T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) in regulation of interleukin(IL)-12 secretion by CD14^+ peripheral blood monocytes from patients with psoriasis vulgaris. Methods From November 2017 to March 2018, a total of 47 patients with psoriasis vulgaris (psoriasis group) and 19 healthy volunteers (control group) were enrolled from Shanxi Provincial People′s Hospital. Peripheral blood mononuclear cells (PBMC) were isolated, and stimulated with Toll-like receptor (TLR) ligands. TIM-3 expression and IL-12 secretion by CD14^+ monocytes were measured by flow cytometry. After blockade of TIM-3 pathway by anti-TIM-3 neutralizing antibody, changes in the downstream signaling pathway molecules in and IL-12 secretion by CD14^+ monocytes were investigated. Two independent samples t-test was used for comparison between two groups, and Pearson correlation test for correlation analysis. Results Under the unstimulated condition, the level of IL-12 secreted by CD14^+ monocytes was very low, and the proportion of CD14^+TIM-3^+ cells was significantly higher in the psoriasis group (12.20%± 2.83%) than in the control group (9.91%± 1.77%, t = 3.270, P = 0.001 7). After the stimulation with TLR ligands, the proportion of CD14^+IL-12^+ cells was significantly lower in the psoriasis group (13.49%± 2.80%) than in the control group (28.97%± 8.97%, t = 10.71, P < 0.000 1), but the proportion of CD14^+TIM-3^+ cells was still higher in the psoriasis group (6.80%± 1.11%) than in the control group (4.85%± 1.37%, t = 6.064, P < 0.000 1). The proportion of CD14^+TIM-3^+ cells was negatively correlated with that of CD14^+IL-12^+ cells in both the control group and psoriasis group (r =-0.473,-0.371 respectively, both P < 0.05). The TIM-3 pathway blockade could induce the decrease of suppressor of cytokine signaling 1 in CD14^+ monocytes, increase the phosphorylation of signaling transducers and activators of transcription 1, and promote IL-12 secretion by CD14^+ monocytes induced by keratinocytes isolated from psoriatic skin lesions. Conclusion TIM-3 plays a crucial role in the negative regulation of CD14^+ monocyte-induced innate immune response in psoriasis.