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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Cloning and analysis of murine cDNA that encodes a fibrogenic lymphokine, fibrosin.
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Cloning and analysis of murine cDNA that encodes a fibrogenic lymphokine, fibrosin.

机译：编码纤维原性淋巴因子纤维蛋白的鼠cDNA的克隆和分析。

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摘要

Tissue fibrosis that complicates chronic inflammation can be a cause of serious morbidity. The molecular links between inflammation and fibrosis appear to be a variety of proteins produced by activated chronic inflammatory cells. Collectively, these fibrogenic cytokines promote the growth of fibroblasts and the production of extracellular matrix that are the characteristic features of fibrotic tissue. In an attempt to clone cDNA for a fibrogenic lymphokine that we had isolated, we transfected COS-7 cells with a cDNA library derived from concanavalin A-stimulated lymphocyte line CDC25. Conditioned medium from the transfected COS-7 cells but not from sham-transfected cells stimulates fibroblast proliferation in vitro. We used heterologous expression in COS-7 cells of pools of CDC25 cDNA and screening for biological activity in conditioned medium to enrich for the cDNA clone(s) that encodes this activity. With this strategy of sib selection we isolated clone 2B3. The culture supernatants of 2B3-transfected COS-7 cells exert maximum growth-stimulating effects on fibroblasts at a dilution of 1:20,000. The isolated cDNA has one open reading frame (216 nucleotides) that has no significant homology with nucleotide sequences that encode other proteins. A synthetic peptide constructed from the deduced amino acid sequence is biologically active in picomolar concentrations, even though it may represent only a portion of the native fibrosin. This lymphokine, which we designate fibrosin, may play a role in regulating fibrogenesis in certain chronic inflammatory diseases.
机译：使慢性炎症复杂化的组织纤维化可能是严重发病的原因。炎症和纤维化之间的分子联系似乎是活化的慢性炎症细胞产生的多种蛋白质。这些成纤维细胞因子共同促进成纤维细胞的生长和细胞外基质的产生，这是纤维化组织的特征。为了克隆我们已经分离的纤维化淋巴因子的cDNA，我们用衍生自伴刀豆球蛋白A刺激的淋巴细胞CDC25的cDNA文库转染了COS-7细胞。来自转染的COS-7细胞但不来自假转染的细胞的条件培养基在体外刺激成纤维细胞增殖。我们使用CDC25 cDNA池的COS-7细胞中的异源表达，并在条件培养基中筛选生物学活性，以富集编码该活性的cDNA克隆。通过这种同胞选择策略，我们分离出克隆2B3。 2B3转染的COS-7细胞的培养上清液以1：20,000的稀释度对成纤维细胞发挥最大的生长刺激作用。分离的cDNA具有一个开放阅读框（216个核苷酸），与编码其他蛋白质的核苷酸序列没有显着同源性。由推导的氨基酸序列构建的合成肽在皮摩尔浓度下具有生物活性，即使它可能仅代表天然纤维蛋白的一部分。我们称其为纤维蛋白的这种淋巴因子可能在某些慢性炎症性疾病中调节纤维生成。

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