生物
分子生物学
重组DNA
互补DNA
刀豆蛋白A
亲和层析
细胞培养
抗血清
干扰素
基因
抗原
生物化学
病毒学
体外
免疫学
酶
遗传学
作者
Kirsten C. Weining,Ursula Schultz,Uwe Münster,Bernd Kaspers,Peter Staeheli
标识
DOI:10.1002/eji.1830261026
摘要
Abstract Supernatants of the chicken T cell line 855 contain antiviral and macrophage‐activating factor activity and strongly activate transcription of the guanylate‐binding protein (GBP) gene in chicken cells. To characterize the cytokine responsible for the GBP‐inducing activity, we chose a cDNA expression cloning strategy in COS cells. Sequencing a positive clone revealed that it encodes chicken interferon‐γ (ChIFN‐γ). Histidine‐tagged ChIFN‐γ was expressed in Escherichia coli and purified by nickel chelate affinity chromatography. ChIFN‐γ from COS cells and E. coli both potently induced GBP RNA synthesis but were rather poor antiviral agents. In macrophages, recombinant ChIFN‐γ strongly stimulated secretion of nitric oxide and enhanced expression of major histocompatibility complex class II antigen. A rabbit antiserum to E. coli ‐derived ChIFN‐γ effectively neutralized the macrophage‐activating factor activity secreted by concanavalin A‐induced spleen cells and various T cell lines, suggesting that IFN‐γ is the major macrophage‐activating factor of the chicken.
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