Airway Smooth Muscle Cells Synthesize Hyaluronan Cable Structures Independent of Inter-α-inhibitor Heavy Chain Attachment
Airway Smooth Muscle Cells Synthesize Hyaluronan Cable Structures Independent of Inter-α-inhibitor Heavy Chain Attachment
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DOI：
10.1074/jbc.m807979200
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发表时间：
2009-02-20
影响因子：
4.8
通讯作者：
Hascall, Vincent C.
中科院分区：
文献类型：
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作者：
Lauer, Mark E.;Fulop, Csaba;Hascall, Vincent C.
间-α-肾上腺素衍生的重链（HC）与透明质酸的共价缔合首先在来自关节炎患者的滑液中描述，并且后来被描述为由许多不同细胞和刺激产生的透明质酸“缆”结构的结构和功能组分。HC转移已显示由TSG-6（肿瘤坏死因子刺激基因6）的蛋白产物介导。考虑到哮喘发作后透明质酸在气道中的积累和随后的白细胞浸润，我们试图表征原代小鼠气道平滑肌细胞（MASM）和原代人气道平滑肌细胞（HASM）中透明质酸“电缆”的HC取代。我们发现，来自缺乏TSG-6的小鼠的细胞在用病毒模拟物poly（I，C）处理时，透明质酸产生或透明质酸介导的白细胞粘附没有缺陷。功能性透明质酸电缆诱导放线菌酮在确认的蛋白质合成的情况下，与或不与聚（I，C）的同时处理。我们表征了其他研究人员用于描述透明质酸电缆的HC-透明质酸复合物的抗体的物种特异性，并发现与来自小鼠和人血清的HC相比，对牛源性HC的亲和力最小。因此，我们在来自这三种来源的血清中培养MASM和HASM细胞，并使用平行的卵丘细胞-卵母细胞复合物（COC）提取物作为阳性对照，分析HC和其他透明质酸结合蛋白的透明质酸提取物。我们的结论是，如果用HC替代来自MASM和HASM细胞的透明质酸电缆，与类似透明质酸质量的COC浸提液相比，替代量显著低于检测限。
The covalent association of inter-alpha-inhibitor-derived heavy chains (HCs) with hyaluronan was first described in synovial fluid from arthritic patients and later described as a structural and functional component of hyaluronan "cable" structures produced by many different cells and stimuli. HC transfer has been shown to be mediated by the protein product of TSG-6 (tumor necrosis factor-stimulated gene 6). Considering the accumulation of hyaluronan in airways following asthmatic attacks and the subsequent infiltration of leukocytes, we sought to characterize HC substitution of hyaluronan "cables" in primary mouse airway smooth muscle cells (MASM) and primary human airway smooth muscle cells (HASM). We found that cells derived from mice lacking TSG-6 had no defect in hyaluronan production or hyaluronan-mediated leukocyte adhesion when treated with the viral mimic poly(I,C). Functional hyaluronan cables were induced by cycloheximide in the confirmed absence of protein synthesis, with or without simultaneous treatment with poly(I, C). We characterized the species specificity of the antibody other investigators used to describe the HC-hyaluronan complex of hyaluronan cables and found minimal affinity to bovine-derived HCs in contrast to HCs from mouse and human sera. Thus, we cultured MASM and HASM cells in serum from these three sources and analyzed hyaluronan extracts for HCs and other hyaluronan-binding proteins, using parallel cumulus cell-oocyte complex (COC) extracts as positive controls. We conclude that, if hyaluronan cables derived from MASM and HASM cells are substituted with HCs, the amount of substitution is significantly below the limit of detection when compared with COC extracts of similar hyaluronan mass.