Analysis of genes isolated from lipopolysaccharide-stimulated rainbow trout (Oncorhynchus mykiss) macrophages
Analysis of genes isolated from lipopolysaccharide-stimulated rainbow trout (Oncorhynchus mykiss) macrophages
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DOI：
10.1016/j.molimm.2004.06.005
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发表时间：
2004-11-01
影响因子：
3.6
通讯作者：
MacKenzie, Simon
中科院分区：
文献类型：
--
作者：
Goetz, Frederick W.;Iliev, Dimitar B.;MacKenzie, Simon
原代细胞培养系统用于获得分化的虹鳟鱼（虹鳟鱼）巨噬细胞，其用大肠杆菌脂多糖（LPS-10 μ g/ml）体外刺激12小时。使用来自LPS刺激的细胞的信使RNA来创建两个cDNA文库，从其中分析总共1048个序列。获得了大量可能与免疫功能相关的cDNA，包括结构蛋白、蛋白酶和抗蛋白酶、转录和翻译调节因子、细胞死亡调节因子、受体、凝集素和免疫球蛋白、细胞因子和趋化因子、细胞表面抗原、信号转导蛋白、抗菌肽和参与类花生酸合成的酶。通过RT-PCR和真实的时间PCR分析并发现被LPS上调的所选基因包括血管细胞粘附分子、CCAAT/增强子结合蛋白β、NF-κ B β抑制剂、CD 209、主要组织相容性II类不变链蛋白、细胞周期蛋白L1、急性期血清淀粉样蛋白A和前列腺素内过氧化物合酶2。(c)2004爱思唯尔有限公司保留所有权利。
A primary cell culture system was used to obtain differentiated rainbow trout (Oncorhynchus mykiss) macrophages that were stimulated with Escherichia coli lipopolysaccharide (LPS-10 mu g/ml) for 12 h in vitro. Messenger RNA from the LPS-stimulated cells was used to create two cDNA libraries from which a total of 1048 sequences were analyzed. A large number of cDNAs were obtained that could be related to immune function including structural proteins, proteases and antiproteases, regulators of transcription and translation, cell death regulators, receptors, lectins and immunoglobulins, cytokines and chemokines, cell surface antigens, signal transduction proteins, antimicrobial peptides, and enzymes involved in eicosanoid synthesis. Selected genes that were analyzed by RT-PCR and real time PCR and found to be upregulated by LPS, included vascular cell adhesion molecule, the CCAAT/enhancer binding protein beta, the inhibitor of NF-kappa B beta, CD209, a major histocompatibility class II-invariant chain protein, cyclin L1, acute phase serum amyloid A, and prostaglandin endoperoxide synthase 2. (c) 2004 Elsevier Ltd. All rights reserved.