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目的:通过比较妊娠不同时期胎盘树突状细胞(dendritic cells,DCs)的前体细胞——单核-巨噬细胞的免疫学特性的差异,探索胎盘免疫细胞及微环境在妊娠耐受和分娩发动中的可能作用。方法:机械法分离中、晚期胎盘中的蜕膜组织细胞,密度梯度离心法提取外周血单个核细胞(PBMC),经CD14+免疫磁珠分选,获得胎盘组织的单核-巨噬细胞。流式细胞仪分析其细胞表型,ELISA检测其细胞培养上清中的细胞因子。再将分离获得的胎盘组织单核-巨噬细胞与同种异体淋巴细胞混合培养,以CCK-8法检测单核-巨噬细胞激发同种异体淋巴细胞增殖的能力,ELISA检测混合培养上清中的细胞因子。结果:自胎盘蜕膜组织中分离获得的单核-巨噬细胞纯度达到93%左右。足月胎盘来源的单核-巨噬细胞低表达与细胞免疫激活有关的细胞表面标志物CD80、CD86、HLA-DR,不表达CD83;细胞培养上清中低表达IL-10、TGF-β;具有较弱的刺激同种异体淋巴细胞增殖的能力,与同种异体淋巴细胞混合培养,上清中低表达IL-10,高表达TGF-β。相比之下,中期妊娠胎盘单核-巨噬细胞CD80、CD86的表达水平较高,以HLA-DR更显著(P<0.05),CD83也有微弱的表达;细胞培养上清中高表达IL-10、TGF-β(P<0.05);其刺激同种异体淋巴细胞增殖的能力也很弱,混合培养上清中高表达IL-10、TGF-β。结论:作为DCs的前体细胞,妊娠不同时期的单核-巨噬细胞在不同的子宫胎盘环境中已处于不同的状态。中期妊娠胎盘来源的单核-巨噬细胞可能通过IL-10发挥免疫抑制功能。
OBJECTIVE: To compare the immunological characteristics of precursor cells - monocytes - macrophages in placental dendritic cells (DCs) during different stages of pregnancy and explore the role of placental immune cells and microenvironment in the development of pregnancy tolerance and childbirth Possible role in launching. Methods: Peripheral blood mononuclear cells (PBMCs) were extracted from the decidual tissue of late placenta by mechanical method and mononuclear macrophages of placenta were obtained by CD14 + immunomagnetic beads sorting. Cell phenotypes were analyzed by flow cytometry and cytokines in cell culture supernatants were assayed by ELISA. The isolated placental mononuclear macrophages were then mixed with allogeneic lymphocytes to detect the proliferation of allogeneic lymphocytes stimulated by monocytes and macrophages by CCK-8 assay. The mixed culture supernatants In the cytokines. Results: The purity of monocyte-macrophage isolated from placental decidua reached about 93%. The expression of CD80, CD86, HLA-DR and CD83 were not significantly expressed in term placenta-derived monocytes - macrophages. IL-10 and TGF-β were down-regulated in the cell culture supernatant. Weak stimulation of allogeneic lymphocyte proliferation ability, mixed with allogeneic lymphocytes culture, low expression of IL-10 in the supernatant, high expression of TGF-β. In contrast, the expression of CD80 and CD86 in placental monocyte-macrophages in the second trimester was higher than that in the HLA-DR (P <0.05), and CD83 was also weakly expressed. In the supernatant of the cell culture, IL-10 , TGF-β (P <0.05). The ability of stimulating allogeneic lymphocyte proliferation was weak. IL-10 and TGF-β were highly expressed in mixed culture supernatant. Conclusion: As precursor cells of DCs, mononuclear macrophages in different stages of pregnancy are in different states in different uterine placenta environment. Placenta-derived mononuclear macrophages in mid-term pregnancy may exert immunosuppressive function through IL-10.