Human Trophoblast-Derived Exosomal Fibronectin Induces Pro-Inflammatory Il-1β Production by Macrophages
Human Trophoblast-Derived Exosomal Fibronectin Induces Pro-Inflammatory Il-1β Production by Macrophages
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DOI：
10.1111/j.1600-0897.2011.00995.x
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发表时间：
2011-10-01
影响因子：
3.6
通讯作者：
Taylor, Douglas D.
中科院分区：
文献类型：
--
作者：
Atay, Safinur;Gercel-Taylor, Cicek;Taylor, Douglas D.
问题我们以前的研究表明，滋养层来源的外泌体诱导巨噬细胞合成和释放促炎细胞因子，包括白细胞介素-1 β（IL-1 β）。本研究的目的是表征的机制和受体与此induction.Method的studyExosomes分离Sw 71滋养层条件培养基通过超滤和超离心。使用从正常供体分离的巨噬细胞，使用细胞松弛素D来阻断外泌体摄取。通过qRT-PCR研究IL-1 β mRNA的诱导，通过蛋白质免疫印迹研究IL-1 β前体蛋白，通过ELISA研究成熟IL-1 β释放。RGD肽用于阻断巨噬细胞α 5 β 1整合素与纤连蛋白的结合。尽管一些细胞因子（如C4 A和CCL 11）的诱导需要摄取，但IL-1 β的诱导在没有外来体内化的情况下发生。细胞松弛素D处理不抑制外泌体介导的IL-1 β mRNA的诱导、前蛋白的产生或成熟IL-1 β的释放。阻断使用RGD肽的纤连蛋白结合表明废除外泌体介导的IL-1 β production.ConclusionAlthough滋养层来源的外泌体已被证明诱导IL-1 β，这是第一次证明IL-1 β诱导外泌体相关的纤连蛋白。基于外泌体相关纤连蛋白的这种促炎作用，它可能代表了一种重要的一般免疫调节机制。
ProblemOur previous studies demonstrated that trophoblast-derived exosomes induced synthesis and release of pro-inflammatory cytokines, including interleukin-1 beta (IL-1 beta) by macrophages. The objective of this study was to characterize the mechanism and receptors associated with this induction.Method of studyExosomes were isolated from Sw71 trophoblast-conditioned media by ultrafiltration and ultracentrifugation. Using macrophages isolated from normal donors, cytochalasin D was used to block exosome uptake. Induction of IL-1 beta mRNA was investigated by qRT-PCR, pro-IL-1 beta protein by western immunoblotting, and mature IL-1 beta release by ELISA. RGD peptides were used to block fibronectin binding by macrophage alpha 5 beta 1 integrin.ResultsUptake of exosomes by macrophages was completely blocked by pretreatment with cytochalasin D. Although induction of some cytokines (such as C4A and CCL11) requires uptake, induction of IL-1 beta occurred without exosome internalization. Cytochalasin D treatment did not inhibit exosome-mediated induction of IL-1 beta mRNA, production of the proprotein, or release of mature IL-1 beta. Blocking of fibronectin binding using RGD peptides demonstrated the abrogation of exosome-mediated IL-1 beta production.ConclusionAlthough trophoblast-derived exosomes have been demonstrated to induce IL-1 beta, this is the first demonstration of IL-1 beta induction by exosome-associated fibronectin. Based on this pro-inflammatory role of exosome-associated fibronectin, it may represent an important general immunoregulatory mechanism.