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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。
Objective To investigate the effect of over-expression of signal transducers and activators of transcription 3 (STAT3) on the anti-differentiation ability of immature dendritic cells (im DCs). METHODS: ImDC cells were isolated and cultured in TNF-α medium. After infection with TNF-α, the cells were infected with pVXX-IRES-Zs Green1-STAT3. Flow cytometry was used to detect the expression of cell surface marker CD86 / MHCⅡ. Western blot was used to detect the expression of IL-10, IDO and NF-κB. The ultrastructural changes of cells were observed by scanning electron microscope (SEM) Cell migration ability. Allogeneic mixed lymphocyte reaction to detect T lymphocyte stimulation. Results Compared with the normal imDC cells, the imDC cells overexpressing STAT3 showed low expression of CD86 / MHCⅡ, but enhanced the expression of NF-κB, IDO and IL-10, When similar, its strong ability to migrate, T lymphocyte stimulating ability was significantly weakened. Conclusion The imDC cells infected with recombinant STAT3 can still maintain the undifferentiated state under the stimulation of TNF-α.