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Structural and functional studies on the human hepatic interleukin-6 receptor. Molecular cloning and overexpression in HepG2 cells.

机译:对人类肝白介素6受体的结构和功能研究。 HepG2细胞中的分子克隆和过度表达。

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摘要

cDNAs coding for the human hepatic interleukin-6 receptor (IL-6-R) have been isolated from a library made from poly(A) RNA of dexamethasone-treated human hepatoma cells (HepG2). We found the hepatic IL-6-R to be identical to the one expressed by leucocytes. A polyclonal antiserum was raised in rabbits against the IL-6-R protein expressed in Escherichia coli. Although the entire IL-6-R protein was used for immunization, only antibodies to the cytoplasmic domain of the IL-6-R were obtained. It is demonstrated by affinity cross-linking and subsequent immunoprecipitation with antibodies against the ligand as well as against the receptor that the cloned cDNA codes for the functional IL-6-R on HepG2 cells. When the hepatic IL-6-R cDNA was overexpressed in HepG2 cells, these cells became more sensitive to low concentrations of IL-6 with respect to the induction of gamma-fibrinogen mRNA.
机译:已从地塞米松处理的人肝癌细胞(HepG2)的poly(A)RNA制备的文库中分离出编码人肝白介素6受体(IL-6-R)的cDNA。我们发现肝IL-6-R与白细胞表达的相同。在兔中产生针对大肠杆菌中表达的IL-6-R蛋白的多克隆抗血清。尽管将整个IL-6-R蛋白用于免疫,但仅获得了针对IL-6-R胞质结构域的抗体。通过亲和性交联和随后用针对配体以及针对受体的抗体进行的免疫沉淀证明,克隆的cDNA编码HepG2细胞上的功能性IL-6-R。当肝IL-6-R cDNA在HepG2细胞中过表达时,这些细胞在诱导γ-纤维蛋白原mRNA方面对低浓度的IL-6更加敏感。

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