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目的:观察妊娠不同阶段胎盘来源的树突样细胞(DC样细胞)刺激脐血淋巴细胞增殖活化特点的差异,进一步研究胎盘免疫细胞在妊娠耐受及分娩发动中的作用。方法:机械分离妊娠中、晚期胎盘中的单个核细胞,用磁珠分离法获取CD14+细胞,以内皮(逆)穿越系统诱导其分化为DC样细胞,并用流式细胞仪分析细胞表型,ELISA法检测细胞培养上清中白细胞介素(IL)-12、IL-10水平,CCK-8法检测其激发脐血淋巴细胞增殖能力,流式细胞术检测其所刺激的脐血T淋巴细胞内细胞因子。结果:足月妊娠胎盘来源的经诱导的细胞高表达与免疫激活有关的细胞表面标志物CD80、CD86;培养上清中检出较高水平的IL-12,极低水平的IL-10,具有较强的刺激脐血淋巴细胞增殖的能力;能刺激脐血T淋巴细胞分化形成较多的IFN-γ产生细胞,而分化形成的IL-10产生细胞则较少。但相比之下,中期妊娠胎盘来源的经诱导的细胞则低表达CD80、CD86(P<0.05),培养上清中检出较高水平的IL-10(P<0.05),但检测出的IL-12浓度极低(P<0.05),刺激脐血淋巴细胞的能力较弱(P<0.05);刺激脐血T淋巴细胞分化形成较多的IL-10产生细胞(P<0.05),而分化形成的IFN-γ产生细胞较少(P<0.05)。结论:妊娠不同时期胎盘来源的单核-巨噬细胞分化形成的DC样细胞免疫学特性不同,高表达CD80、CD86、IL-12的DC样细胞细胞在妊娠免疫耐受中止及分娩发动中发挥着一定的作用,而低表达CD80、CD86,高表达IL-10的细胞可能参与了妊娠过程中的免疫耐受。
Objective: To observe the differences of placenta-derived dendritic cells (DC-like cells) in different stages of pregnancy in stimulating proliferation and activation of cord blood lymphocytes and to further investigate the role of placental immune cells in the development of pregnancy tolerance and labor. Methods: Mononuclear cells from placenta of middle and late pregnancy were isolated by mechanical separation. CD14 + cells were isolated by magnetic bead separation and differentiated into DC-like cells by endothelial (reverse) cross system. Cell phenotype was analyzed by flow cytometry. ELISA The levels of interleukin (IL) -12 and IL-10 in the supernatant of the cells were detected by the method of CCK-8. The proliferation of cord blood lymphocytes was detected by CCK-8 assay. Cytokines. RESULTS: Placenta-derived induced cells in full-term pregnancy expressed high levels of cell surface markers CD80 and CD86 related to immune activation. Higher levels of IL-12 and extremely low levels of IL-10 were detected in culture supernatants with Strong ability to stimulate the proliferation of cord blood lymphocytes; can stimulate the differentiation of cord blood T lymphocytes to form more IFN-γ-producing cells, and differentiated IL-10-producing cells are less. In contrast, the placenta-derived induced cells in the second trimester had a lower expression of CD80 and CD86 (P <0.05), and a higher level of IL-10 (P <0.05) was detected in the culture supernatant. However, The IL-12 concentration was very low (P <0.05), and the ability of stimulating cord blood lymphocytes was weaker (P <0.05). T cells of cord blood were stimulated to differentiate to form more IL-10 producing cells (P <0.05) Differentiation resulted in fewer IFN-γ producing cells (P <0.05). CONCLUSION: DC-like cells differentiated from placenta-derived mononuclear macrophages at different stages of pregnancy exhibit different immunological characteristics. DC-like cells highly expressing CD80, CD86, and IL-12 play an important role in the termination of immunotolerance and the onset of labor However, the low expression of CD80, CD86, high expression of IL-10 cells may be involved in the process of immune tolerance during pregnancy.