Identification of soluble interleukin-4 receptor in rat glomerular epithelial cells
Identification of soluble interleukin-4 receptor in rat glomerular epithelial cells
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DOI：
10.1016/s0167-4889(99)00117-2
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发表时间：
1999-10-13
影响因子：
5.1
通讯作者：
Maruyama, N
中科院分区：
文献类型：
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作者：
Chen, GP;Nagasawa, R;Maruyama, N
白细胞介素4（IL-4）是一种与肾小球疾病有关的多效性细胞因子，受膜结合型IL-4 R（mIL-4 R）的正调节，受可溶性IL-4 R（sIL-4 R）的负调节。由于天然sIL-4 R仅在小鼠中有记录，因此我们在大鼠中进行了这项研究，以确定它们是否也表达sIL-4 R，特别是在肾细胞中。为此目的设计了一对IL-4 R引物，并用于聚合酶链反应。结果表明，大鼠脾细胞和肾小球上皮细胞（GEC）中均有sIL-4 R表达。序列分析表明，大鼠sIL-4 R的mRNA有一段75 bp的插入序列。该插入片段在跨膜区上游产生终止TGA密码子，导致形成sIL-4 R。随后筛选的肾脏cDNA文库，使我们能够获得全长3605 bp的sIL-4 R的cDNA，全长3530 bp的mIL-4 R的cDNA也被确定为一个更长的序列比以前发表的。在IL-4 R阳性的39个克隆中，2个被证实为sIL-4 R，37个被证实为mIL-4 R。然后，将sIL-4 R cDNA的翻译部分构建到表达载体中，使我们能够获得重组sIL-4 R-myc融合蛋白。应用该重组sIL-4 R，证实sIL-4 R能拮抗IL-4诱导的脾细胞增殖。本研究证实sIL-4 R在肾细胞中表达，并具有拮抗作用。(C)1999 Elsevier Science B. V.保留所有权利。
Interleukin (IL)-4, a pleiotropic cytokine involved in many glomerular diseases, is regulated positively by membrane-bound IL4R (mIL-4R) and negatively by soluble IL-4R (sIL4R). Because natural sIL-4R has been documented only in mice, we undertook this study in rats to determine whether they, too, express sIL-4R, particularly in kidney cells. A pair of IL-4R primers was designed for this purpose and used in the polymerase chain reaction. As a result, sIL-4R was found not only in rats spleen cells but also in their glomerular epithelial cells (GEC). Sequence analysis revealed that the mRNA of rat sIL-4R has a 75-bp insert sequence. This insert generated a termination TGA codon upstream from the transmembrane region, resulting in formation of the sIL-4R. Subsequent screening of the kidney cDNA library enabled us to obtain the whole 3605-bp cDNA of sIL-4R; the full-length 3530-bp mIL-4R cDNA was also identified as a much longer sequence than previously published. Among the total 39 clones positive for IL-4R, two were confirmed as sIL-4R, and 37 clones were positive for mIL-4R. Next, the translated portion of sIL-4R cDNA was constructed into an expression vector, enabling us to obtain a recombinant sIL4R-myc fusion protein. By using this recombinant sIL-4R, we proved that sIL-4R can antagonize the IL-4-induced proliferation of spleen cells. Present study demonstrated that sIL-4R is expressed in kidney cells and antagonistically functional. (C) 1999 Elsevier Science B.V. All rights reserved.