Innate Immunity in Human Embryonic Stem Cells: Comparison with Adult Human Endothelial Cells
Innate Immunity in Human Embryonic Stem Cells: Comparison with Adult Human Endothelial Cells
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DOI：
10.1371/journal.pone.0010501
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发表时间：
2010-05-05
期刊：
影响因子：
3.7
通讯作者：
Mitchell, Jane A.
中科院分区：
文献类型：
--
作者：
Foeldes, Gabor;Liu, Alexander;Mitchell, Jane A.
用人胚胎干细胞（hESC）衍生的细胞治疗人类疾病现已接近现实，但对它们对生理和病理损伤的反应知之甚少。细胞通过激活Toll样受体（TLR）进行应答的能力在大多数组织的先天免疫感知中至关重要，但也扩展到更普遍的危险感知，例如，免疫应答。G.心肌细胞中的氧化应激。我们使用生物标志物释放和基因阵列分析来比较hESC分化前后的反应，以及原代人内皮细胞的反应。通过免疫染色、FACS分选和心肌细胞的搏动活性，在分化培养物中证实了心肌细胞和内皮细胞的存在。未分化的hESC不响应于CXCL 8对革兰氏阳性或革兰氏阴性细菌的释放，或针对TLR 1-9的一系列PAMP（病原体相关分子模式）（除了鞭毛蛋白，TLR 5的激活剂）。令人惊讶的是，在包括心肌细胞和内皮细胞的培养物中，在hESC分化的4个月内维持缺乏TLR依赖性应答。相反，原代培养的人主动脉内皮细胞（HAEC）表现出广泛的PAMP的反应。在hESC中证实了下游TLR信号传导途径的表达，并且绕过TLR的IL-1 β、TNF α和INF γ刺激CXCL 8释放。NF κ B通路在hESC中也有表达，NF κ B能够转位到细胞核。在hESC中检测到TLR的低表达水平，特别是TLR 1和4，解释了hESC对主要TLR信号缺乏应答。TLR 5水平在分化的hESC和HAEC之间是相似的，并且TLR 5的siRNA敲低消除了对鞭毛蛋白的应答。这些发现对移植的hESC衍生细胞的存活和功能具有潜在的影响。
Treatment of human disease with human embryonic stem cell (hESC)-derived cells is now close to reality, but little is known of their responses to physiological and pathological insult. The ability of cells to respond via activation of Toll like receptors (TLR) is critical in innate immune sensing in most tissues, but also extends to more general danger sensing, e. g. of oxidative stress, in cardiomyocytes. We used biomarker release and gene-array analysis to compare responses in hESC before and after differentiation, and to those in primary human endothelial cells. The presence of cardiomyocytes and endothelial cells was confirmed in differentiated cultures by immunostaining, FACS-sorting and, for cardiomyocytes, beating activity. Undifferentiated hESC did not respond with CXCL8 release to Gram positive or Gram negative bacteria, or a range of PAMPs (pathogen associated molecular patterns) for TLRs 1-9 (apart from flagellin, an activator of TLR5). Surprisingly, lack of TLR-dependent responses was maintained over 4 months of differentiation of hESC, in cultures which included cardiomyocytes and endothelial cells. In contrast, primary cultures of human aortic endothelial cells (HAEC) demonstrated responses to a broad range of PAMPs. Expression of downstream TLR signalling pathways was demonstrated in hESC, and IL-1 beta, TNF alpha and INF gamma, which bypass the TLRs, stimulated CXCL8 release. NF kappa B pathway expression was also present in hESC and NFkB was able to translocate to the nucleus. Low expression levels of TLRs were detected in hESC, especially TLRs 1 and 4, explaining the lack of response of hESC to the main TLR signals. TLR5 levels were similar between differentiated hESC and HAEC, and siRNA knockdown of TLR5 abolished the response to flagellin. These findings have potential implications for survival and function of grafted hESC-derived cells.