Department of Biomedical Sciences, University of Guelph, Guelph, Ontario, Canada.
Immunol Cell Biol. 2010 Jan;88(1):63-71. doi: 10.1038/icb.2009.62. Epub 2009 Sep 15.
Spontaneous early and mid-gestation fetal losses occur in swine. At both stages, endometrial lymphocytes associated with smaller, paler conceptuses have fewer pro-angiogenic and more pro-inflammatory cytokine transcripts compared with robust conceptuses. We hypothesized that similar differences occur in conceptus-associated dendritic cells (DCs). Using laser capture-microdissection, dendritic cell-specific intercellular adhesion molecule-grabbing non-integrin (DC-SIGN)(+) cells were isolated from attachment sites of healthy and arresting conceptuses at gestation day (gd)20 and 50. DC-SIGN(+) cells were screened using real-time PCR for vascular endothelial cell growth factor (Vegf), its receptors, semaphorins (Sema) and plexins (Plxn), and for toll-like receptor (Tlr) transcripts to address potential activation pathways. Homogenized endometrial and trophoblast biopsies were quantified for type 1/type 2 cytokine transcripts/proteins. DC-SIGN(+) cells from healthy and arresting conceptuses had more Vegf transcripts at early than mid gestation whereas transcripts for Vegfr1 and Vegfr2 were stable. In gd20 arresting site DC-SIGN(+) cells, Neuropilin-2 transcripts were elevated, whereas at gd50 arresting sites, Plxn-A2 increased and Sema3A transcripts were lost. Tlr-1, Tlr-4 and Tlr-6 transcript abundance was independent of conceptus health. At gd20, type 1 cytokines were prevalent, whereas at gd50 type 2 cytokines predominated in endometrium and trophoblast. Thus, gestational features, characteristic of haemochorial placentation, are present in species with distinctly different placentation.
自发性早中期妊娠胎儿丢失发生在猪中。在这两个阶段,与较小、较苍白的胚胎相关的子宫内膜淋巴细胞的促血管生成细胞因子转录物较少,促炎细胞因子转录物较多,与强壮的胚胎相比。我们假设在与胚胎相关的树突状细胞(DC)中也存在类似的差异。使用激光捕获显微切割,从妊娠第 20 天和第 50 天的附着部位分离出健康和停滞胚胎的细胞间粘附分子抓取非整联蛋白(DC-SIGN)(+)细胞。使用实时 PCR 对 DC-SIGN(+)细胞进行血管内皮细胞生长因子(Vegf)、其受体、信号素(Sema)和丛蛋白(Plxn)以及 Toll 样受体(TLR)转录物进行筛选,以解决潜在的激活途径。将匀浆的子宫内膜和滋养层活检用于 1 型/2 型细胞因子转录物/蛋白的定量。与中期妊娠相比,健康和停滞胚胎的早期 DC-SIGN(+)细胞的 Vegf 转录物更多,而 Vegfr1 和 Vegfr2 的转录物则稳定。在 gd20 停滞部位的 DC-SIGN(+)细胞中,神经纤毛蛋白-2 转录物升高,而在 gd50 停滞部位,Plxn-A2 增加,Sema3A 转录物丢失。TLR-1、TLR-4 和 TLR-6 转录物丰度与胚胎健康无关。在 gd20 时,1 型细胞因子占主导地位,而在 gd50 时,子宫内膜和滋养层以 2 型细胞因子为主。因此,具有明显不同胎盘形成的物种存在与绒毛膜胎盘形成特征一致的妊娠特征。