Abbadi Amina, Lauer Mark, Swaidani Shadi, Wang Aimin, Hascall Vincent
From the Department of Biomedical Engineering and Department of Chemistry, Cleveland State University, Cleveland, Ohio 44115.
From the Department of Biomedical Engineering and.
J Biol Chem. 2016 Jan 15;291(3):1448-55. doi: 10.1074/jbc.M115.704288. Epub 2015 Nov 24.
Many cells, including murine airway epithelial cells, respond to a variety of inflammatory stimuli by synthesizing leukocyte-adhesive hyaluronan (HA) cables that remain attached to their cell surfaces. This study shows that air-liquid interface cultures of murine airway epithelial cells (AECs) also actively synthesize and release a majority of their HA onto their ciliated apical surfaces to form a heavy chain hyaluronan (HC-HA) matrix in the absence of inflammatory stimuli. These matrices do not resemble the rope-like HA cables but occur in distinct sheets or rafts that can capture and embed leukocytes from cell suspensions. The HC-HA modification involves the transfer of heavy chains from the inter-α-inhibitor (IαI) proteoglycan, which has two heavy chains (HC1 and HC2) on its chondroitin sulfate chain. The transesterification transfer of HCs from chondroitin sulfate to HA is mediated by tumor necrosis factor-induced gene 6 (TSG-6), which is up-regulated in inflammatory reactions. Because the AEC cultures do not have TSG-6 nor serum, the source of IαI, assays for HCs and TSG-6 were done. The results show that AECs synthesize TSG-6 and their own heavy chain donor (pre-IαI) with a single heavy chain 3 (HC3), which are also constitutively expressed by human renal proximal tubular epithelial cells. These leukocyte adhesive HC3-HA structures were also found in the bronchoalveolar lavage of naïve mice and were observed on their apical ciliated surfaces. Thus, these leukocyte-adhesive HA rafts are now identified as HC3-HA complexes that could be part of a host defense mechanism filling some important gaps in our current understanding of murine airway epithelial biology and secretions.
许多细胞,包括小鼠气道上皮细胞,通过合成仍附着在其细胞表面的白细胞粘附透明质酸(HA)电缆来响应各种炎症刺激。本研究表明,小鼠气道上皮细胞(AECs)的气液界面培养物在没有炎症刺激的情况下,也会主动合成并将大部分HA释放到其纤毛顶端表面,以形成重链透明质酸(HC-HA)基质。这些基质与绳索状的HA电缆不同,而是以独特的片状或筏状形式出现,能够捕获并嵌入细胞悬液中的白细胞。HC-HA修饰涉及来自α-抑制因子(IαI)蛋白聚糖的重链转移,该蛋白聚糖在其硫酸软骨素链上有两条重链(HC1和HC2)。HCs从硫酸软骨素到HA的酯交换转移由肿瘤坏死因子诱导基因6(TSG-6)介导,TSG-6在炎症反应中上调。由于AEC培养物中没有TSG-6和血清(IαI的来源),因此进行了HCs和TSG-6的检测。结果表明,AECs合成TSG-6及其自身的重链供体(前IαI),其具有单条重链3(HC3),人肾近端小管上皮细胞也组成性表达这些物质。在未接触过抗原的小鼠的支气管肺泡灌洗物中也发现了这些白细胞粘附性HC3-HA结构,并在其顶端纤毛表面观察到。因此,这些白细胞粘附性HA筏现在被鉴定为HC3-HA复合物,它们可能是宿主防御机制的一部分,填补了我们目前对小鼠气道上皮生物学和分泌物理解中的一些重要空白。