过表达STAT3对小鼠未成熟树突状细胞分化的影响

The effect of STAT3 overexpression on the differentiation of mouse immature dendritic cells

目的研究过表达信号转导与转录活化因子3(signal transducers and activators of transcription 3,STAT3)对未成熟树突细胞(immature dendritic cells,im DC)抗分化能力的影响。方法分离获取imDC细胞,分别加入TNF-α培养、空载病毒感染后加入TNF-α培养、p LVX-IRES-Zs Green1-STAT3慢病毒感染后加入TNF-α培养。流式细胞术鉴定细胞表面分子标志物CD86/MHCⅡ的表达变化,Western blot检测细胞免疫耐受因子IL-10、IDO、NF-κB的表达变化;扫描电镜观察细胞超微结构变化,Transwell实验检测细胞迁移能力。同种异体混合淋巴细胞反应检测对T淋巴细胞的刺激能力。结果与正常分化的imDC细胞相比,过表达STAT3的imDC细胞在TNF-α刺激后表面标志分子CD86/MHCⅡ低表达,NF-κB、IDO、IL-10蛋白的表达增强,细胞形态与未分化时相似,其迁移能力较强、T淋巴细胞刺激能力明显减弱。结论经感染重组STAT3病毒的im DC细胞在TNF-α刺激下,仍能保持未分化状态。

This study was designed to explore whether STAT3 can increase the ability of anti-differentiation in imDC. imDC isolated from mice were infected by pLVX -IRES-ZsGreenl-STAT3 lentivirus, and then TNF-α was added into the culture. The expression of surface molecules markers CD86/MHC Ⅱ were checked by flow cytometry Western blot was used to detect the expression of IL-10/IDO/NF-κB; scanning electron microscopy was applied to observe uhrastructure changes of imDC; while transwell assay was used to test the cell migration of imDC. The ability of stimulating T lymphocytes was detected by mixed lymphocyte reaction. Data showed that compared with normal imDC cells, the imDC cells overexpressing STAT3 after TNF-α stimulation demonstrated significant lower expression of CD86/MHC Ⅱ, higher expression of STAT3/IL-10/IDO/NF-κB, strengthened migration ability, and weakened ability of T lymphocyte stimulation. The above results suggest that overexpression of STAT3 can increase anti-differentiation ability of imDC.