【摘要】 目的本研究旨在探讨滋养细胞对T淋巴细胞细胞因子产生和平衡的影响,以及滋养细胞对T淋巴细胞特异性转录因子的影响。方法分离早孕胎盘细胞滋养细胞,收集滋养细胞培养上清液为滋养细胞条件培养基,供淋巴细胞培养用。分离健康未孕妇女的外周血单个核细胞,用人T细胞富集柱纯化CD3+T淋巴细胞。以佛波酯(PMA)刺激T淋巴细胞,酶联免疫吸附分析法测定淋巴细胞产生的细胞因子浓度,实时定量聚合酶链反应检测T淋巴细胞中Th1、Th2和Th17免疫特异性转录因子水平。结果滋养细胞条件培养基显著减少T淋巴细胞产生IL-2、IFN-γ和TNF-α,IL-4和IL-10的水平无统计学差异,而Th1/Th2(IL-2/IL-4、IFN-γ/IL-4、TNF-α/IL-4、IL-2/IL-10、IFN-γ/IL-10和TNF-α/IL-10)显著降低;滋养细胞条件培养基显著抑制T淋巴细胞产生IL-17。滋养细胞条件培养基显著降低T淋巴细胞Th1特异性转录因子STAT-4的表达,显著增强Th2特异性转录因子STAT-6和GATA-3表达;但对Th1特异性转录因子T-bet、Th17特异性转录因子RORC无显著性影响。结论滋养细胞主要通过抑制T淋巴细胞Th1细胞因子产生而使Th1/Th2平衡向Th2漂移,并且抑制Th17免疫,在妊娠免疫耐受中起着重要的作用。更多还原

【Abstract】 Objective To clarify the effect of placental trophoblasts on T lymphocyte by observing the alteration in the production of cytokines and the expression of specific transcription factors for Th1,Th2 and Th17 immunity in T lymphocyte.Methods Placental trophoblasts were isolated from chorionic villi of normal pregnancy and conditioned medium was made after 72 hours culture of trophoblast.PBMC were isolated from healthy female donors and T lymphocytes were purified and cultured in the presence or absence of conditioned medium.Enzyme- linked sorbent immune assay(ELISA) was used to detect the concentration of IL- 2,TNF- γ,IFN- α,IL- 4,IL- 10 and IL- 17 in supernatants of T cell culture and real- time PCR was used to detect the expression of specific transcription factors for Th1 immunity(T- bet and STAT- 4),Th2(GATA- 3 and STAT- 6) and Th17(RORC) in T lymphocyte.Results The level of IL- 2,IFN- γ,TNF- α and IL- 17 was significantly decreased when the T lymphocytes were cultured in conditioned medium compared with control medium,while the level of IL- 10 and IL- 4 were comparable.The presence of conditioned medium decreased the ratio of Th1 / Th2 as indexed by IL- 2 / IL- 4,IFN- γ / IL- 4,TNF- α / IL- 4、IL- 2 / IL- 10、IFN- γ / IL- 10 and TNF- α / IL- 10 compared with control medium.The expression of GATA- 3 and STAT- 6 were significantly increased and STAT- 4 was reduced when T cells were cultured in conditioned medium,while the expression of T- bet and RORC were comparable.Conclusion Placental trophoblast- induced shift of Th1 / Th2 balance toward Th2 and inhibition of Th17 might be among the mechanisms involved in maternal tolerance to fetus.更多还原