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目的:探讨Toll受体1/2(Toll like receptor1/2,TLR1/2)信号对荷瘤小鼠来源的CD8~+T细胞功能的影响及其可能机制。方法:利用小鼠Lewis肺癌细胞株3LL建立小鼠肺癌荷瘤模型,MACS分选小鼠脾CD8~+T细胞;体外经PBS或TLR1/2激动剂BLP刺激后,Real-time PCR和流式细胞术分别从基因和蛋白水平检测CD8~+T细胞的TLR分子表达;用ELISA和流式细胞术检测经PBS或BLP刺激后的CD8~+T细胞分泌细胞因子和增殖的能力,并用关键信号分子抑制剂分析可能的分子机制。结果:与PBS对照组相比,TLR1/2激动剂BLP不但有效上调荷瘤机体CD8~+T细胞TLR1和TLR2分子的基因水平[TLR1:(0.353±0.015)vs(0.101±0.017),P<0.01;TLR2:(0.232±0.031)vs(0.080±0.004),P<0.05]及蛋白水平(P<0.05),而且显著促进CD8~+T细胞分泌功能性细胞因子[IFN-γ:(2 375±305)vs(850±50),P<0.05;IL-2:(1 600±200)vs(350±50),P<0.05]和增殖的能力(P<0.05),这一效应依赖于NF-KB和P38通路。结论:TLR1/2信号直接作用于荷瘤小鼠的CD8~+T细胞并促进其功能,该研究既丰富了TLR的作用范围,也为基于TLR激动剂的肿瘤生物治疗提供了实验依据。
Objective: To investigate the effect of Toll Receptor 1/2 (TLR1 / 2) signaling on the function of CD8 + T cells derived from tumor-bearing mice and its possible mechanism. Methods: Mouse lung cancer tumor model was established by 3LL mouse Lewis lung carcinoma cell line, and splenic CD8 ~ + T cells were isolated by MACS. After stimulated by PBS or TLR1 / 2 agonist BLP, Real-time PCR and flow cytometry Cytokines were used to detect the expression of TLRs on CD8 ~ + T cells, respectively. The ability of CD8 + T cells stimulated by PBS or BLP to secrete cytokines and proliferation was detected by ELISA and flow cytometry. Molecular Inhibitors Analysis of Possible Molecular Mechanisms. Results: Compared with PBS control group, TLR1 / 2 agonist BLP not only effectively upregulated the gene level of TLR1 and TLR2 in TLR1: (0.353 ± 0.015) vs (0.101 ± 0.017), P < 0.01); TLR2: (0.232 ± 0.031) vs (0.080 ± 0.004), P <0.05] and protein level (P <0.05), and significantly promoted the secretion of functional cytokines [ ± 305) vs (850 ± 50), P <0.05; IL-2: (1 600 ± 200) vs (350 ± 50), P <0.05] and proliferative ability (P < NF-KB and P38 pathways. CONCLUSION: TLR1 / 2 signal acts directly on CD8 + T cells of tumor-bearing mice and promotes its function. This study not only enriches the scope of TLR but also provides an experimental basis for tumor biological therapy based on TLR agonists.