CLONING AND ANALYSIS OF MURINE CDNA THAT ENCODES A FIBROGENIC LYMPHOKINE, FIBROSIN
CLONING AND ANALYSIS OF MURINE CDNA THAT ENCODES A FIBROGENIC LYMPHOKINE, FIBROSIN
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DOI：
10.1073/pnas.92.6.2154
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发表时间：
1995-03-14
影响因子：
11.1
通讯作者：
WYLER, DJ
中科院分区：
文献类型：
--
作者：
PRAKASH, S;ROBBINS, PW;WYLER, DJ
慢性炎症并发的组织纤维化可导致严重的疾病。炎症和纤维化之间的分子联系似乎是由激活的慢性炎症细胞产生的各种蛋白质，总的来说，这些成纤维细胞因子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。为了克隆我们分离的一种纤维性淋巴因子的cDNA，我们用从豆豆蛋白a刺激的淋巴细胞系CDC25中获得的cDNA文库转染COS-7细胞。从横切的COS-7细胞而不是从假转染的细胞中提取的条件培养基刺激成纤维细胞的体外增殖。我们在COS-7细胞中异种表达CD625 cDNA，并在条件培养基中筛选生物活性，以富集编码该活性的cDNA克隆。采用sib选择策略分离克隆2B3，转染2B3的COS-7细胞培养上清液在1:20 000稀释时对成纤维细胞的生长刺激作用最大。分离的cDNA有一个开放的阅读框（216个核苷酸），与编码其他蛋白质的核苷酸序列没有显著的同源性。从推导出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，即使它可能只代表天然纤维蛋白的一部分。这种淋巴因子，我们称之为纤维蛋白，可能在某些慢性炎症疾病的纤维生成中起调节作用。
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 transected 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 CD625 cDNA and screening for biological activity in conditioned medium to enrich for the cDNA clone(s) that encodes this activity. With iis 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.