TLR8激动剂对胃癌患者树突细胞表型及功能的调节作用

目的研究TLR8激动剂对胃癌患者树突细胞体外培养的影响。方法应用血细胞分离机无菌采集肿瘤患者外周血,实验室分离诱导为树突细胞和T细胞,加入TLR8激动剂处理未成熟的树突细胞刺激成熟。Western blotting检测树突细胞中pNF-κB的表达水平,流式检测树突细胞表面标记分子表达,ELISA检测对照组和激动剂组IL-12、TNF-α的表达水平,树突细胞与自体T淋巴细胞共培养后流式检测T细胞表面标记分子表达。结果 TLR8激动剂刺激后树突细胞中pNF-κB表达增加,树突细胞表面标记HLR-DR、CD11c、CD54、CD80、CD83、CD86分子水平上调,激动剂组IL-12、TNF-α的表达水平升高,共培养后T细胞表面标记CD3、CD28分子无明显变化。结论 TLR8激动剂体外刺激促进胃癌患者的树突细胞成熟,为TLR8激动剂作为树突细胞疫苗佐剂提供证据。

Regulatory effects of TLR8 agonist on the phenotype and function of dendritic cells in patients with gastric cancer

To investigate the effect of TLR8 agonist on the culture of dendritic cells form gastric cancer, the peripheral blood of tumor patients were aseptically collected by blood cell separator, then dendritic cells and T cells were induced by laboratory isolation. TLR8 agonists were added to treat immature dendritic cells for maturation. The expression level of pNF-κB in dendritic cells was detected by Western blotting;the expression of surface marker molecules on dendritic cells were detected by flow cytometry. ELISA was used to detect the expression levels of IL-12 and TNF-α in the control and agonist groups. The expression of T cell surface marker molecules were detected through co-culture of dendritic cells and autologous T lymphocytes. Data showed that the expression of pNF-κB was increased in dendritic cells after TLR8 agonist stimulation, furthermore, TLR8 agonist treatment also up-regulated the levels of HLR-DR, CD11 c, CD54, CD80, CD83 and CD86 on the surface of dendritic cells, as well as the levels of IL-12 and TNF-α, but CD3 and CD28 molecules labeled on the surface of T cells showed no significant changes after co-culture. Taken together, TLR8 agonist stimulation promotes dendritic cell maturation in gastric cancer patients and provides evidence for TLR8 agonists as dendritic cell vaccine adjuvant.