Distribution of decidual natural killer cells and macrophages in the neighbourhood of the trophoblast invasion front: a quantitative evaluation
Distribution of decidual natural killer cells and macrophages in the neighbourhood of the trophoblast invasion front: a quantitative evaluation
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DOI：
10.1093/humrep/det353
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发表时间：
2014-01-01
影响因子：
6.1
通讯作者：
Sedlmayr, P.
中科院分区：
文献类型：
--
作者：
Helige, C.;Ahammer, H.;Sedlmayr, P.
蜕膜自然杀伤（dNK）细胞和蜕膜巨噬细胞（dMph）是否在滋养层浸润前沿附近富集？形态学图像分析和面积细胞密度计算，排除观察者的偏见，显示了丰富的蜕膜白细胞在附近的滋养层invasion front.In以前的研究中，蜕膜白细胞的数量是目测计数中或高功率字段。然而，这些方法不能揭示白细胞和浸润性滋养层细胞之间的确切空间关系，因此容易出现主观误差。因此，需要一种更客观的方法。应用一种新的形态测量图像分析方法，在来自妊娠早期胎盘形成部位的人类组织碎片（n 7）以及在妊娠早期蜕膜组织与同一妊娠的胎盘绒毛的共培养物（n 15）中研究了白细胞群体，所述共培养物代表滋养层侵入的适当体外模型。孕早期蜕膜组织获自在孕龄710周时进行选择性终止妊娠的妇女。对组织切片进行双重化学染色，一方面用于dNK细胞或dMph的标志物，另一方面用于侵袭性绒毛外滋养层细胞。为了分析白细胞的分布，定义了不同的细胞区室以及细胞邻近区域。最后，计算相对面积细胞密度，并将这些数据与滋养层侵袭的体外模型以及来自无滋养层细胞的壁蜕膜的组织碎片的数据进行比较。在妊娠早期胎盘形成部位，在靠近侵袭性滋养层的位置发现较高密度的dNK细胞以及dMph（P 0.01），与排除滋养层的组织中蜕膜白细胞的平均面细胞密度相比。白细胞的最高面积细胞密度被确定为距离滋养层细胞20 m，而在更远的区域，它甚至低于平均水平，表明这些白细胞向滋养层入侵前沿迁移。然而，在三维共培养模型中，我们发现了dMph的富集（P 0.01），但dNK细胞无明显变化（P 0，形态学图像分析依赖于无背景和交叉免疫组化染色的强免疫组化染色。该方法不仅可用于复发性流产的研究，而且可用于肿瘤免疫学和炎症领域。由欧盟委员会支持（胚胎植入控制卓越网络（EMBIC），FP 6 -512040，首席研究员：P.S.），和奥地利格拉兹医科大学Franz Lanyar基金会（347号赠款）。没有一位作者声明存在利益冲突。
Do decidual natural killer (dNK) cells and decidual macrophages (dMph) become enriched in the vicinity of the trophoblast invasion front?Morphometric image analysis and areal cell density calculations, which excluded observer bias, showed an enrichment of decidual leukocytes in the neighbourhood of the trophoblast invasion front.In previous studies, the number of decidual leukocytes was visually counted in medium- or high power fields. These methods, however, cannot reveal the exact spatial relationship between leukocytes and invasive trophoblast cells, and are therefore prone to subjective errors. Thus, a more objective approach is required.Applying a new method of morphometric image analysis, leukocyte populations were studied in human tissue fragments derived from first trimester placentation sites (n 7) as well as in co-cultures of first trimester decidual tissue with placental villi of the same pregnancy representing an appropriate in vitro model of trophoblast invasion (n 15).First trimester decidual tissue was obtained from women undergoing elective terminations of pregnancy at 710 weeks of gestational age. Tissue sections were double-stained immunohistochemically for markers of dNK cells or dMph on one hand, and for invasive extravillous trophoblast cells on the other. To analyse the distribution of leukocytes, distinct cell compartments as well as cell neighbourhood areas were defined. Finally, relative areal cell densities were calculated and these data were compared with those of an in vitro model of trophoblast invasion as well as with tissue fragments derived from decidua parietalis without trophoblast cells.At first trimester placentation sites, a higher density of dNK cells as well as of dMph was found in close proximity to the invasive trophoblast (P 0.01), compared with the average areal cell density of decidual leukocytes in the tissue with exclusion of the trophoblast. The highest areal cell density of leukocytes was determined up to a distance of 20 m from the trophoblast cells, whereas in more distant regions it was even lower than average, indicating a migration of these leukocytes towards the trophoblast invasion front. In the three-dimensional co-culture model, however, we found an enrichment of dMph (P 0.01) but not of dNK cells (P 0,05) in the neighbourhood of the invasive trophoblast.The morphometric image analysis depends on intense immunohistochemical staining that is free of background and cross-reactivity.The presented method will be useful not only for the investigation of recurrent miscarriage but also in the fields of tumour immunology and inflammation.The study was supported by the European Commission (Network of Excellence The Control of Embryo Implantation (EMBIC), FP6-512040, lead researcher: P.S.), and by the Franz Lanyar Foundation of the Medical University of Graz, Austria (Grant 347). None of the authors declared a conflict of interests.