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目的:观察不同妊娠阶段胎盘源性树突样细胞刺激T细胞活化增殖特点的差异,为分析胎盘免疫细胞在妊娠耐受和分娩发动中的可能作用提供线索。方法:机械分离中、晚期胎盘中的单核-巨噬细胞,以内皮逆穿越系统诱导其分化为树突状细胞(DC)。将不同妊娠阶段胎盘单核-巨噬细胞来源的DC样细胞与同种异体T细胞共育,以CCK-8法检测其激发同种异体T细胞增殖能力及其刺激同种异体T细胞所产生的胞内细胞因子,并用流式细胞仪分析DC样细胞表型。结果:足月胎盘来源的单核-巨噬细胞经过内皮单层正逆双向穿越诱导培养后,细胞出现树突状形态改变,并高表达与树突状细胞免疫激活有关的细胞表面标志物CD80、CD86、HLA-DR。足月妊娠胎盘单核-巨噬细胞来源DC样细胞具有较强的刺激同种异体淋巴细胞增殖的能力,同种异体淋巴细胞活化时细胞内IFN-γ+亚型的表达十分明显,但几乎不出现IL-10+亚型。但相比之下,中期妊娠胎盘单核-巨噬细胞来源的DC样细胞刺激同种异体T细胞增殖的能力却很弱(P<0.05),被刺激的同种异体T细胞中CD8-(即CD4+)亚群IFN-γ+细胞显著减少(P<0.05),而CD8-和CD8+亚群中IL-10+细胞亚群却显著增多。对细胞的表型分析结果显示,中期妊娠胎盘单核-巨噬细胞来源的DC样细胞CD80、CD86及HLA-DR的表达均显著低下(P<0.05)。结论:不同妊娠时期胎盘单核-巨噬细胞分化形成具有免疫激活作用的树突状细胞的能力不同,提示其生物学特性有所不同,其可能与妊娠耐受和分娩发动有关。
OBJECTIVE: To observe the differences of the characteristics of T cell activation and proliferation stimulated by placenta-derived dendritic cells in different gestation stages and provide clues for the analysis of the possible role of placental immune cells in the development of pregnancy tolerance and childbirth. METHODS: Monocyte-macrophages in the late placenta were mechanically isolated and induced to differentiate into dendritic cells (DCs) by a reverse endothelial crossing system. The DC-like cells derived from placental mononuclear macrophages in different stages of pregnancy were co-cultured with allogeneic T cells. The proliferative ability of allogeneic T cells and their effect on allogenic T cells were detected by CCK-8 assay Of intracellular cytokines and analyzed for DC-like cell phenotype by flow cytometry. Results: The mononuclear macrophages derived from full-term placenta were induced to undergo dendritic morphological changes through both positive and negative transversal endothelial monolayers, and high expression of cell surface marker CD80, which is related to dendritic cell immune activation , CD86, HLA-DR. The placental monocyte-macrophage-derived DCs in term pregnancy have a strong ability of stimulating the proliferation of allogeneic lymphocytes. The expression of intracellular IFN-γ + subtypes is very evident when allogeneic lymphocytes are activated, but almost Does not appear IL-10 + subtype. However, in contrast, DC-like cells derived from placental monocyte-macrophages in the second trimester showed a weak ability of stimulating the proliferation of allogeneic T cells (P <0.05). Among the stimulated allogeneic T cells, CD8- ( (CD4 +) subpopulation IFN-γ + cells (P <0.05), while the CD8- and CD8 + subpopulations of IL-10 + cell subpopulations were significantly increased. Phenotypic analysis of the cells showed that the expression of CD80, CD86 and HLA-DR in monocyte-macrophage-derived DCs in the second trimester of pregnancy were significantly lower (P <0.05). CONCLUSION: Different abilities of placental monocyte-macrophage to differentiate into dendritic cells with immune activation during different pregnancy stages suggest different biological characteristics, which may be related to pregnancy tolerance and labor initiation.