Bidarimath Mallikarjun, Khalaj Kasra, Kridli Rami T, Wessels Jocelyn M, Koti Madhuri, Tayade Chandrakant
Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON, Canada, K7L 3N6.
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada, N1G 2W1.
Cell Tissue Res. 2016 Dec;366(3):747-761. doi: 10.1007/s00441-016-2470-2. Epub 2016 Aug 8.
Chemokines play a significant role in pregnancy, especially during embryonic attachment and placental development. During early pregnancy, immune cells are recruited extensively to the endometrium in several species including pigs. However, this recruitment is solely mediated by the presence of the conceptus in pigs making it a unique feature compared with other species (humans, primates and mice). To understand the biological significance of chemokine expression and immune cell recruitment in the context of fetal loss, we investigate a well-characterized porcine fetal loss model during the window of early pregnancy at gestational day (gd) 20 and mid-pregnancy (gd50). These periods coincide with 25-40 % of conceptus loss. Using targeted quantitative polymerase chain reaction and Western blot approaches, we screened a specific set of chemokines. Comparisons were made with endometrial lymphocytes (ENDO LY), endometrium and chorioallantoic membranes (CAM) associated with spontaneously arresting and healthy conceptus attachment sites (CAS). mRNA expression studies revealed an increased expression of CXCR3 and CCR5 in ENDO LY and of CXCL10, CXCR3, CCL5 and CCR5 in the endometrium associated with arresting CAS at gd20. DARC was decreased in the endometrium at gd50. CCL1 was increased in CAM associated with arresting CAS at gd50. Some of these differences were also noted at the protein level (CXCL10, CXCR3, CCL5 and CCR5) in the endometrium and CAM. CD45+ immunohistochemistry demonstrated a significantly higher localization in ENDO LY in the endometrium associated with healthy versus arresting counterparts. Most of these differences were observed in early pregnancy and might contribute towards a shift in immune cell functions.
趋化因子在妊娠过程中发挥着重要作用，尤其是在胚胎着床和胎盘发育期间。在妊娠早期，包括猪在内的几种物种的子宫内膜会大量募集免疫细胞。然而，在猪中这种募集仅由孕体的存在介导，这使其与其他物种（人类、灵长类动物和小鼠）相比具有独特之处。为了了解趋化因子表达和免疫细胞募集在胎儿丢失背景下的生物学意义，我们研究了一个特征明确的猪胎儿丢失模型，该模型处于妊娠早期第20天和妊娠中期（第50天）的窗口期。这些时期与25%-40%的孕体丢失相吻合。我们使用靶向定量聚合酶链反应和蛋白质印迹方法筛选了一组特定的趋化因子。将与自然停止发育和健康孕体着床部位（CAS）相关的子宫内膜淋巴细胞（ENDO LY）、子宫内膜和绒毛尿囊膜（CAM）进行了比较。mRNA表达研究显示，在第20天与停止发育的CAS相关的ENDO LY中，CXCR3和CCR5的表达增加，在子宫内膜中CXCL10、CXCR3、CCL5和CCR5的表达增加。在第50天，子宫内膜中的DARC减少。在第50天与停止发育的CAS相关的CAM中，CCL1增加。其中一些差异在子宫内膜和CAM的蛋白质水平（CXCL10、CXCR3、CCL5和CCR5）上也有体现。CD45+免疫组织化学显示，与健康对照相比，子宫内膜中与停止发育的对照相关的ENDO LY中的定位显著更高。这些差异大多在妊娠早期观察到，可能有助于免疫细胞功能的转变。
