论文部分内容阅读
为研究活化诱导的凋亡淋巴细胞来源的DNA(凋亡DNA)对小鼠巨噬细胞的体外活化作用,用ConA活化小鼠脾脏淋巴细胞并诱导其凋亡,将AnnexinV+凋亡细胞来源的DNA定义为凋亡DNA,而正常淋巴细胞来源的DNA定义为正常DNA,分别与小鼠巨噬细胞株RAW264.7体外孵育48 h,然后用碘化丙啶(PI)染色方法观察细胞吞噬的DNA平均荧光强度(MFI),流式细胞仪测定细胞表面分子的表达,ELISA检测II型干扰素(IFN-γ)、白细胞介素12(IL-12)和肿瘤坏死因子(TNF-α)的分泌情况。结果显示:凋亡DNA和正常DNA都能被RAW264.7有效地吞噬;然而,凋亡DNA较正常DNA能显著上调RAW264.7细胞表面MHC II类分子以及协同刺激分子CD40、CD80、CD86的表达,同时能够诱导细胞分泌高水平的IFN-γ、IL-12和TNF-α。表明正常DNA对巨噬细胞无活化作用,而凋亡DNA对巨噬细胞有很强的活化作用,可上调巨噬细胞的APC功能并促使其分泌多种细胞因子。提示凋亡的自身DNA作为免疫原,可有效活化巨噬细胞。
To investigate the in vitro activation of murine macrophages by activation-induced apoptosis of lymphocyte-derived DNA (apoptotic DNA), ConA-activated mouse spleen lymphocytes and induce apoptosis, AnnexinV + apoptotic cell-derived DNA Defined as apoptotic DNA, while normal lymphocyte-derived DNA was defined as normal DNA and incubated with mouse macrophage cell line RAW264.7 for 48 h in vitro, and then phagocytosed with propidium iodide (PI) staining method The mean fluorescence intensity (MFI), the expression of cell surface molecules by flow cytometry, the secretion of type II interferon (IFN-γ), interleukin 12 (IL-12) and tumor necrosis factor (TNF- Happening. The results showed that both apoptotic DNA and normal DNA were effectively phagocytosed by RAW264.7. However, apoptotic DNA significantly up-regulated the expression of MHC class II molecules and costimulatory molecules CD40, CD80 and CD86 on RAW264.7 cells compared with normal DNA , At the same time be able to induce cells to secrete high levels of IFN-γ, IL-12 and TNF-α. The results showed that normal DNA had no activation on macrophages, whereas apoptotic DNA had a strong activation on macrophages, which could upregulate APC function of macrophages and induce its secretion of various cytokines. Suggest that the apoptotic DNA itself can effectively activate macrophages.