Cloning and biologic activities of a bovine interferon-alpha isolated from the epithelium of a rotavirus-infected calf
Cloning and biologic activities of a bovine interferon-alpha isolated from the epithelium of a rotavirus-infected calf
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DOI：
10.1089/jir.1996.16.25
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发表时间：
1996-01-01
影响因子：
2.3
通讯作者：
Collins, RA
中科院分区：
文献类型：
--
作者：
Chaplin, PJ;Entrican, G;Collins, RA
编码独特的牛 (Be) 干扰素 (IFN) α（称为 BoIFN-α E）的 cDNA 是从感染轮状病毒的小牛中分离的肠上皮细胞产生的。 BoIFN-α E cDNA 序列与其他 BoIFN-α 亚型具有大于 90% 的同一性。编码 BolFN-α E 的 cDNA 已使用杆状病毒苜蓿银纹夜蛾核型多角体病毒 (AcMNPV) 作为载体在昆虫细胞中表达。感染重组病毒的昆虫细胞将相对分子质量为 19,500 的蛋白质分泌到培养基中，而在感染野生型 AcMNPV 的细胞中未观察到。从感染编码 IFN-α E 的重组 AcMNPV 的昆虫细胞培养物中收获的上清液抑制了 Semliki 森林病毒在牛细胞系中的复制，通常显示 10(6) 然而，重组BoIFN-α E 和BoIFN-α(1) 1 对WC1(+) γ/δ T 细胞增殖的活性存在差异，纯化的(> 99%) WC1(+) γ/δ T 细胞未能增殖为IFN-α(1) 1 或刀豆球蛋白A IFN-α E 对这些细胞起到微弱的增殖信号作用，证明了两种密切相关的 BoIFN-α 亚型之间的功能差异。
A cDNA encoding a distinct bovine (Be) interferon (IFN) alpha, designated BoIFN-alpha E, was generated from gut epithelial cells isolated from a rotavirus-infected calf. The BoIFN-alpha E cDNA sequence shared a greater than 90% identity with the other BoIFN-alpha subtypes, The cDNA encoding BolFN-alpha E has been expressed in insect cells using the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) as a vector. Insect cells infected with recombinant virus secreted a protein with a relative molecular mass of 19,500 into the culture medium not observed in cells infected with wild-type AcMNPV, Supernatants harvested from cultures of insect cells infected with the recombinant AcMNPV encoding IFN-alpha E inhibited the replication of Semliki Forest virus in a bovine cell line and typically showed 10(6) dilution units/ml of antiviral activity, However, differences were observed between the activities of recombinant BoIFN-alpha E and BoIFN-alpha(1) 1 on the proliferation of WC1(+) gamma/delta T cells, Purified (> 99%) WC1(+) gamma/delta T cells failed to proliferate to IFN-alpha(1) 1 or concanavalin A and IFN-alpha E acted as a weak proliferative signal to these cells, demonstrating a functional difference between two closely related BoIFN-alpha subtypes.