Constitutive Expression of Pentraxin 3 ( PTX3) Protein by Human Amniotic Membrane Cells Leads to Formation of the Heavy Chain ( HC)-Hyaluronan ( HA)-PTX3 Complex
Constitutive Expression of Pentraxin 3 ( PTX3) Protein by Human Amniotic Membrane Cells Leads to Formation of the Heavy Chain ( HC)-Hyaluronan ( HA)-PTX3 Complex
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DOI：
10.1074/jbc.m113.525287
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发表时间：
2014-05-09
影响因子：
4.8
通讯作者：
Tseng, Scheffer C. G.
中科院分区：
文献类型：
--
作者：
Zhang, Suzhen;Zhu, Ying-Ting;Tseng, Scheffer C. G.
背景：来自羊膜的HC-HA是由羊膜细胞产生的。结果：5-羟色胺3与纯化的AM HC-HA紧密结合，由AM细胞结构性分泌，形成HC-HA-PTX3复合体。结论：PTX3是AM HC-HA-PTX3复合体的重要组成部分。意义：由AM内源性产生的HC-HA-PTX3可能在胎儿发育过程中发挥重要的保护作用，并可作为一种积极的治疗药物。重链(HC)-透明质酸(HA)是从人羊膜(AM)中提纯的、由胰酶间抑制剂(II)产生的HCl与透明质酸(HA)共价连接形成的复合体，负责AM的抗炎、抗瘢痕和抗血管生成作用这种HC-HA复合体是由肿瘤坏死因子刺激的基因6的结构性表达和AM细胞内源性产生II产生的。五角蛋白3(PTX3)是一种典型的长五角蛋白，在针对选定病原体的先天性免疫中发挥着非多余的作用，也有助于稳定HC-HA以确保女性生育。我们注意到在AM上皮和致密间质中PTX3呈强阳性染色。PTX3由培养的AM上皮细胞和间质细胞组成性表达和分泌，并受肿瘤坏死因子和白介素1的显著上调。用琼脂糖包被捕获含有HA的基质，通过Western印迹分析，尽管在血清和肿瘤坏死因子存在的情况下培养，但由AM细胞而不是人皮肤成纤维细胞形成了HC-HA-PTX3复合体。然而，外源性PTX3帮助人皮肤成纤维细胞形成具有琼脂糖层的HC-HA-PTX3复合体。此外，PTX3可以与抗人II抗体包被琼脂糖包被的AM细胞提取物中的HC-HA复合体免疫共沉淀。这种HC-HA-PTX3复合体可以在体外重组，并表现出与已报道的AM HC-HA-PTX3对M2巨噬细胞极化的作用相似的效果。在4M GnHCl存在下，PTX3与AM HC-HA之间的紧密结合经受住了四次CsCl超离心。这些结果表明，PTX3作为AM HC-HA-PTX3复合体的组成部分由AM细胞结构性表达和分泌，参与AM HC-HA-PTX3的生物学功能。
Background: HC-HA from the amniotic membrane (AM) is produced by AM cells. Results: Pentraxin 3 tightly binds to purified AM HC-HA and is constitutively secreted by AM cells, leading to HC-HA-PTX3 complex formation. Conclusion: PTX3 is an integral component of AM HC-HA-PTX3 complexes. Significance: HC-HA-PTX3, produced endogenously by the AM, may play an important protective role during fetal development and can be an active therapeutic agent.Heavy chain (HC)-hyaluronan (HA), a complex formed by the covalent linkage between HC1 from the inter--trypsin inhibitor (II) and HA, purified from the human amniotic membrane (AM), is responsible for the anti-inflammatory, antiscarring, and antiangiogenic actions of the AM. This HC-HA complex is produced by constitutive expression of TNF-stimulated gene 6 and endogenous production of II by AM cells. Pentraxin 3 (PTX3), a prototypic long pentraxin that plays a non-redundant role in innate immunity against selected pathogens, also helps stabilize HC-HA to ensure female fertility. Here we noted strong positive PTX3 staining in the AM epithelium and compact stroma. PTX3 was constitutively expressed and secreted by cultured AM epithelial and stromal cells and, further, greatly up-regulated by TNF and IL-1. Using an agarose overlay to trap the HA-containing matrix, the HC-HA-PTX3 complex was formed, as analyzed by Western blot analysis, by AM cells but not human skin fibroblasts, despite being cultured in the presence of serum and TNF. However, exogenous PTX3 helps human skin fibroblasts form the HC-HA-PTX3 complex with an agarose overlay. Furthermore, PTX3 can be coimmunoprecipitated with the HC-HA complex from agarose-overlaid AM cell extracts by an anti-human II antibody. Such a HC-HA-PTX3 complex can be reconstituted in vitro and exhibit similar effects as those reported for AM HC-HA-PTX3 on polarization of M2 macrophages. The tight binding between PTX3 and AM HC-HA withstands four runs of CsCl ultracentrifugation in the presence of 4 m GnHCl. These results indicate that PTX3 is constitutively expressed and secreted by AM cells as an integral component of the AM HC-HA-PTX3 complex and contributes to the biological function of AM HC-HA-PTX3.