Roussev R G, Higgins N G, McIntyre J A
Methodist Center for Reproduction and Transplantation Immunology, Methodist Hospital of Indiana, Indianapolis 46202.
J Reprod Immunol. 1993 Sep;25(1):15-29. doi: 10.1016/0165-0378(93)90039-k.
The placenta is a rich source of immunocompetent cells. We have studied the phenotype, number and origin of placental mononuclear blood cells isolated from 32 normal term placentae using 4 color flow cytometry. Respective maternal and cord blood leucocyte preparations were also compared. Placental tissue without extraembryonic membranes was cut into small pieces and divided. One portion was washed extensively with ice-cold PBS. Both tissue portions were disrupted in a blender and cells were dissociated by using a 180 mu sieve. Leucocytes were isolated by Ficoll-Hypaque density gradient centrifugation. Maternal and cord bloods were HLA typed and in cases of HLA-A2 or B7/40 disparity, monoclonal anti-HLA antibodies to these antigens showed that unwashed placental tissue contained 35% maternal and 65% fetal cells. This ratio, however, was not reflected for a given cell phenotype. In comparison, washed placental tissue contained cells of fetal origin only. Both unwashed and washed placental tissue contained fewer CD3 and CD4, but more CD8 cells than maternal and cord blood. Markers of NK cells such as, CD16, CD56, and CD57 showed this cellular phenotype to be 15 times more abundant in the placental preparations than in cord and maternal blood. The quantitative differences between peripheral blood and placental CD8 and NK cells were further explored with an antiprogesterone receptor antibody in combination with anti-CD8, anti-CD57 and anti-HLA-DR. The number of progesterone receptor (PGR) positive cells was three times higher in placental tissues than in cord or maternal blood. These data indicate that the phenotypic frequencies of certain placental leucocytes are significantly different from maternal and fetal peripheral blood. Progesterone and the presence of PGR may be important in the differential retention of placental leucocytes.
胎盘是免疫活性细胞的丰富来源。我们使用四色流式细胞术研究了从32例足月正常胎盘中分离出的胎盘单个核血细胞的表型、数量和来源。还比较了相应的母体和脐带血白细胞制剂。将没有胎膜的胎盘组织切成小块并分开。一部分用冰冷的PBS广泛洗涤。将两部分组织在搅拌机中破碎，并用180目筛网解离细胞。通过Ficoll-Hypaque密度梯度离心分离白细胞。对母体和脐带血进行HLA分型，在HLA-A2或B7/40不一致的情况下，针对这些抗原的单克隆抗HLA抗体显示，未洗涤的胎盘组织含有35%的母体细胞和65%的胎儿细胞。然而，对于给定的细胞表型，该比例并未得到体现。相比之下，洗涤后的胎盘组织仅含有胎儿来源的细胞。未洗涤和洗涤后的胎盘组织中CD3和CD4细胞均比母体和脐带血中的少，但CD8细胞更多。NK细胞标志物，如CD16、CD56和CD57显示，这种细胞表型在胎盘制剂中的丰度是脐带血和母体血中的15倍。用抗孕激素受体抗体联合抗CD8、抗CD57和抗HLA-DR进一步探究外周血与胎盘CD8和NK细胞之间的定量差异。胎盘组织中孕激素受体（PGR）阳性细胞的数量比脐带血或母体血中的高三倍。这些数据表明，某些胎盘白细胞的表型频率与母体和胎儿外周血有显著差异。孕激素和PGR的存在可能在胎盘白细胞的差异性滞留中起重要作用。
