Coumans B, Thellin O, Zorzi W, Melot F, Bougoussa M, Melen L, Zorzi D, Hennen G, Igout A, Heinen E
Institute of Human Histology, University of Liège, Belgium.
J Immunol Methods. 1999 Apr 22;224(1-2):185-96. doi: 10.1016/s0022-1759(99)00021-6.
To test the hypothesis that CD95-L (Fas-L) present on trophoblastic cells plays a part in establishing foeto-placental tolerance by inducing apoptosis of immune defence cells, we cocultured trophoblasts with lymphoid cells and scored the frequency of cell death in these cultures. We prepared human trophoblastic cells from term placentas removed by C-section and placed them in culture for 48 h before introducing the lymphoid cells. We added Jurkat cells, a CD3 + lymphoid cell line, or purified T cells from human blood to the cultured trophoblasts and monitored apoptosis by electron microscopy and flow cytometry after TUNEL or annexin V labelling. The frequency of cell death in the CD3 + cell population was higher when the lymphoid cells were cocultured with trophoblastic cells than when they were cultured alone. This frequency increased with time but was reduced when anti-CD95-L antibodies were added to the culture medium. Cell death was less frequent in the lymphoid cell population when trophoblasts were replaced with human fibroblasts not expressing CD95-L.
为了验证滋养层细胞上存在的CD95-L(Fas-L)通过诱导免疫防御细胞凋亡在建立胎儿-胎盘耐受性中发挥作用这一假说,我们将滋养层细胞与淋巴细胞共培养,并对这些培养物中的细胞死亡频率进行了评分。我们从剖宫产取出的足月胎盘中制备人滋养层细胞,并在引入淋巴细胞之前将其培养48小时。我们将Jurkat细胞(一种CD3 +淋巴细胞系)或从人血中纯化的T细胞添加到培养的滋养层细胞中,并在TUNEL或膜联蛋白V标记后通过电子显微镜和流式细胞术监测细胞凋亡。当淋巴细胞与滋养层细胞共培养时,CD3 +细胞群体中的细胞死亡频率高于单独培养时。该频率随时间增加,但当向培养基中添加抗CD95-L抗体时降低。当用不表达CD95-L的人成纤维细胞替代滋养层细胞时,淋巴细胞群体中的细胞死亡频率较低。