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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。
Objective This study aimed to investigate the effects of trophoblasts on the production and balance of cytokines in T lymphocytes and the effects of trophoblasts on T lymphocyte-specific transcription factors. Methods Placental trophoblast cells from early pregnancy were isolated and the trophoblast culture supernatants were collected as trophoblast conditioned medium for lymphocyte culture. Peripheral blood mononuclear cells from healthy, non-pregnant women were isolated and CD3 + T lymphocytes were purified using a human T-cell enrichment column. T lymphocytes were stimulated with phorbol myristate (PMA), cytokines produced by lymphocytes were assayed by enzyme-linked immunosorbent assay, and the levels of Th1, Th2 and Th17 immunospecific transcription factors in T lymphocytes were detected by real-time quantitative polymerase chain reaction. Results Trophoblast conditioned medium significantly reduced the levels of IL-2, IFN-γ and TNF-α produced by T lymphocytes, but no significant difference was observed in the levels of Th1 / Th2 (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) Inhibit T lymphocytes from producing IL-17. Trophoblast conditioned medium significantly reduced the expression of Th1-specific transcription factor STAT-4 and significantly increased the expressions of Th2-specific transcription factors STAT-6 and GATA-3. However, Th1-specific transcription factors T-bet and Th17 Sex transcription factor RORC had no significant effect. Conclusion Trophoblasts mainly play an important role in the immune tolerance of pregnancy by inhibiting the Th1 cytokine production of T lymphocytes, leading to Th2 / Th2 balance shift to Th2 and inhibiting Th17 immunity.