The expression and activity of Toll-like receptors in the preimplantation human embryo suggest a new role for innate immunity.
The expression and activity of Toll-like receptors in the preimplantation human embryo suggest a new role for innate immunity.
复制标题
DOI：
10.1093/humrep/deab188
复制
发表时间：
2021-09-18
期刊：
影响因子：
--
通讯作者：
Brison DR
中科院分区：
文献类型：
--
作者：
Aboussahoud WS;Smith H;Stevens A;Wangsaputra I;Hunter HR;Kimber SJ;Seif MW;Brison DR
先天免疫系统在人类早期胚胎发育过程中是否活跃？模式识别受体和先天免疫 Toll 样受体 (TLR) 基因在植入前人类胚胎中广泛表达，并且该途径似乎对 TLR 配体有活跃的反应。早期人类胚胎对其当地环境高度敏感，然而，人们对胚胎如何检测和响应特定环境线索知之甚少。虽然已知母体免疫反应是植入时妊娠建立的关键，但人类胚胎检测和发出病原体存在信号的能力尚不清楚。通过分析已发表的转录组数据（n = 40）来评估人类胚胎中TLR家族和相关基因的表达。第5天（D-5）人类胚胎（n=25）在已知TLR配体存在下培养，并与对照相比测量基因表达和细胞因子产生。超出治疗需求的人类胚胎是在几个 ART 中心的知情同意下捐赠的。将胚胎在 TLR3 和 TLR5 配体 Poly (I:C) 和鞭毛蛋白存在下培养至第 6 天 (D-6)，通过定量 PCR 测量基因表达，并使用细胞计数珠阵列测量细胞因子释放到培养基中。 TLR 和相关基因，包括下游信号分子，在人类胚胎发育的所有阶段都有不同的表达。结果显示，TLR 9 和 5 在囊胚中表达最强，TLR 9、5、2、6 和 7 在整个发育过程中表达最强。用 TLR3 和 TLR5 配体 Poly (I: C) 和鞭毛蛋白刺激第 5 天囊胚，导致 TLR 基因 mRNA 表达水平发生变化，包括透明质酸介导的运动受体 (HMMR)、TLR5、TLR7、核因子活化 B 细胞 (NF-κB) 和单核细胞趋化蛋白 1 (MCP-1) 的 kappa 轻链增强子（与未刺激对照相比，P < 0.05，P < 0.001），并释放到细胞因子和趋化因子的培养基中，特别是 IL8（P = 0.00005 和 0.01277）分别为鞭毛蛋白和聚 (I: C))。这是一项描述性实验研究，表明 TLR 系统在人类胚胎中活跃并能够发挥作用，但并未证实任何特定作用。尽管我们鉴定了一系列 TLR 基因的胚胎转录本，但在已发表的研究中，表达模式并不总是一致，并且某些基因的表达水平较低，因此有可能这些基因是从母体基因组而不是胚胎基因组表达的。这是关于人类胚胎中先天免疫 TLR 系统的许多成分的表达和活性的首次报告。了解 TLR 在植入前人类发育过程中的作用可能对于揭示自然受孕和 ART 期间胚胎质量和妊娠起始的免疫机制和潜在临床标志物很重要。这项工作由高等教育部、利比亚国、英国医学研究委员会、NIHR 地方综合研究网络和 NIHR 曼彻斯特临床研究设施以及欧盟 Horizon 2020 研究和创新计划根据 Marie Skłodowska-Curie 赠款协议第 812660 号 (DohART-NET) 资助。根据 H2020 规则，欧盟资助的研究活动不会牺牲任何新的人类胚胎，这些活动仅涉及记录的延时和转录组数据集的计算机分析。所有作者都没有任何需要声明的利益冲突。不适用。
Is the innate immunity system active in early human embryo development? The pattern recognition receptors and innate immunity Toll-like receptor (TLR) genes are widely expressed in preimplantation human embryos and the pathway appears to be active in response to TLR ligands. Early human embryos are highly sensitive to their local environment, however relatively little is known about how embryos detect and respond to specific environmental cues. While the maternal immune response is known to be key to the establishment of pregnancy at implantation, the ability of human embryos to detect and signal the presence of pathogens is unknown. Expression of TLR family and related genes in human embryos was assessed by analysis of published transcriptome data (n = 40). Day 5 (D-5) human embryos (n = 25) were cultured in the presence of known TLR ligands and gene expression and cytokine production measured compared to controls. Human embryos surplus to treatment requirements were donated with informed consent from several ART centres. Embryos were cultured to Day 6 (D-6) in the presence of the TLR3 and TLR5 ligands Poly (I: C) and flagellin, with gene expression measured by quantitative PCR and cytokine release into medium measured using cytometric bead arrays. TLR and related genes, including downstream signalling molecules, were expressed variably at all human embryo developmental stages. Results showed the strongest expression in the blastocyst for TLRs 9 and 5, and throughout development for TLRs 9, 5, 2, 6 and 7. Stimulation of Day 5 blastocysts with TLR3 and TLR5 ligands Poly (I: C) and flagellin produced changes in mRNA expression levels of TLR genes, including the hyaluronan-mediated motility receptor (HMMR), TLR5, TLR7, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and monocyte chemoattractant Protein-1 (MCP-1) (P < 0.05, P < 0.001 compared to unstimulated controls), and release into culture medium of cytokines and chemokines, notably IL8 (P = 0.00005 and 0.01277 for flagellin and Poly (I: C), respectively). This was a descriptive and experimental study which suggests that the TLR system is active in human embryos and capable of function, but does not confirm any particular role. Although we identified embryonic transcripts for a range of TLR genes, the expression patterns were not always consistent across published studies and expression levels of some genes were low, leaving open the possibility that these were expressed from the maternal rather than embryonic genome. This is the first report of the expression and activity of a number of components of the innate immunity TLR system in human embryos. Understanding the role of TLRs during preimplantation human development may be important to reveal immunological mechanisms and potential clinical markers of embryo quality and pregnancy initiation during natural conception and in ART. This work was funded by the Ministry of Higher Education, The State of Libya, the UK Medical Research Council, and the NIHR Local Comprehensive Research Network and NIHR Manchester Clinical Research Facility and the European Union’s Horizon 2020 Research and Innovation Programmes under the Marie Skłodowska-Curie Grant Agreement No. 812660 (DohART-NET). In accordance with H2020 rules, no new human embryos were sacrificed for research activities performed from the EU funding, which concerned only in silico analyses of recorded time-lapse and transcriptomics datasets. None of the authors has any conflict of interest to declare. n/a.
登录
查看更多内容
影响因子：
6.1
作者：
Fazeli, A;Bruce, C;Anumba, DO
通讯作者：
Anumba, DO
影响因子：
64.8
作者：
Alexopoulou, L;Holt, AC;Flavell, RA
通讯作者：
Flavell, RA
影响因子：
3.5
作者：
Dobson, AT;Raja, R;Pera, RAR
通讯作者：
Pera, RAR
影响因子：
6.2
作者：
Carpentier, PA;Begolka, WS;Miller, SD
通讯作者：
Miller, SD
影响因子：
3.8
作者：
Deb, K;Chaturvedi, MM;Jaiswal, YK
通讯作者：
Jaiswal, YK