Chaplin P J, Entrican G, Gelder K I, Collins R A
Division of Immunology and Pathology, Institute for Animal Health, Compton, Newbury, Berkshire.
J Interferon Cytokine Res. 1996 Jan;16(1):25-30. doi: 10.1089/jir.1996.16.25.
A cDNA encoding a distinct bovine (Bo) 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 BoIFN-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.
从感染轮状病毒的小牛分离的肠道上皮细胞中产生了一种编码独特牛（Bo）干扰素（IFN）α的cDNA，命名为BoIFN-αE。BoIFN-αE的cDNA序列与其他BoIFN-α亚型的序列同一性大于90%。编码BoIFN-αE的cDNA已利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向培养基中分泌了一种相对分子质量为19,500的蛋白质，而感染野生型AcMNPV的细胞中未观察到这种现象。从感染编码IFN-αE的重组AcMNPV的昆虫细胞培养物中收获的上清液抑制了塞姆利基森林病毒在牛细胞系中的复制，通常显示出10^6稀释单位/毫升的抗病毒活性。然而，观察到重组BoIFN-αE和BoIFN-α11对WC1+γ/δT细胞增殖的活性存在差异。纯化的（>99%）WC1+γ/δT细胞对IFN-α11或伴刀豆球蛋白A不发生增殖反应，而IFN-αE对这些细胞起弱增殖信号作用，这表明两种密切相关的BoIFN-α亚型之间存在功能差异。
