Expression of the complement alternative pathway by human myoblasts in vitro: Biosynthesis of C3, factor B, factor H and factor I
Expression of the complement alternative pathway by human myoblasts in vitro: Biosynthesis of C3, factor B, factor H and factor I
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DOI：
10.1002/eji.1830251238
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发表时间：
1995-12-01
影响因子：
5.4
通讯作者：
Fontaine, M
中科院分区：
文献类型：
--
作者：
Legoedec, J;Gasque, P;Fontaine, M
在这项研究中，我们证明了正常人成肌细胞和人横纹肌肉瘤细胞系CRL1558和HTB153在体外表达补体替代途径成分C3、因子B、因子H和因子I。通过蛋白质印迹分析和生物合成标记检测培养物上清液中的蛋白质，然后进行免疫沉淀实验，并通过 ELISA 进行定量。新分泌的蛋白质在结构和功能上与其血清对应物相似。检测到另外一种具有 H 因子免疫反应性的 43 kDa 多肽，该多肽可能对应于血浆中发现的 N 端截短形式。通过逆转录酶-聚合酶链式反应分析将蛋白质表达与mRNA表达相关联。补体旁路途径C3、因子B和因子H的主要蛋白质由骨骼肌细胞以50至150ng/10(6)细胞/ml的速率组成型产生，因子I的表达率为20ng/10(6)细胞/ml。这些体外合成受到炎症细胞因子的调节。 γ 干扰素显着上调 C3、B 因子和 H 因子的表达，但对 I 因子的产生没有影响。 Interleukin-1β强烈增强C3和B因子的产生，而对I因子和H因子的分泌有弱增强作用或没有作用。人类成肌细胞系构成了一个有趣的模型来分析人类骨骼肌细胞的补体生物合成。体内骨骼肌的局部补体表达可能与某些肌肉炎症或病理过程有关。
In this study, we demonstrate expression in vitro of complement alternative pathway components C3, factor B, factor H and factor I by normal human myoblasts and human rhabdomyosarcoma cell lines CRL1558 and HTB153. Proteins in culture supernatants were detected by Western (protein) blot analysis and biosynthetic labeling followed by immunoprecipitation experiments, and quantified by ELISA. Newly secreted proteins were structurally and functionally similar to their serum counterparts. An additional polypeptide of 43 kDa with factor H immunoreactivity was detected, which could correspond to the N-terminal truncated form found in plasma. Protein expression was correlated with mRNA expression by reverse transcriptase-polymerase chain reaction analysis. The major proteins of complement alternative pathway C3, factor B and factor H were produced constitutively by skeletal muscle cells at a rate of 50 to 150 ng/10(6) cells/ml and factor I was expressed 20 ng/10(6) cells/ml. These syntheses in vitro were regulated by inflammatory cytokines. Interferon-gamma significantly up-regulated C3, factor B and factor H expression, but had no effect on factor I production. Interleukin-1 beta strongly enhanced C3 and factor B production and had a weak enhancing or no effect on factor I and factor H secretion. Human myoblast cell lines constitute an interesting model to analyze complement biosynthesis by human skeletal muscle cells. Local complement expression by skeletal muscle in vivo may be implicated in some muscular inflammatory or pathological processes.