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The 3D8 single chain variable fragment protein suppress infectious bronchitis virus transmission in the transgenic chickens

机译:3D8单链可变片段蛋白抑制转基因鸡中的传染性支气管炎病毒传播

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摘要

Infectious bronchitis (IB) generated by the infectious bronchitis virus (IBV) causes economic difficulties for livestock farmers. The 3D8 single chain variable fragment (scFv) protein is a recombinant antibody with nuclease activity that shows antiviral effects against various DNA and RNA viruses in mice and chickens. In this experiment, 3D8 scFv G transgenic chickens produced by crossing 3D8 scFv G transgenic rooster and wild type hens were screened by genomic PCR and immunohistochemistry analysis. 3D8 scFv transgenic chickens, wild type sibling chickens, and SPF chickens were directly infected with IBV (5 chickens per group) and indirectly infected by airborne propagation (15 chickens per group). The relative IBV shedding titers were measured by quantitative real-time PCR using oropharyngeal and cloacal swabs on days 3 and 5 after intraocular infection. The viral load was significantly decreased in the 3D8 scFv transgenic chickens from the contact transmission group. Additionally, blood was collected from each group on day 17 post-infection. The ELISA results showed a marked reduction of the antibody titer against IBV in the 3D8 scFv transgenic chickens from the contact transmission group. These results suggest that the 3D8 scFv protein potentially inhibits infectious bronchitis virus transmission in chickens.
机译:由传染性支气管炎病毒(IBV)产生的传染性支气管炎(IB)给牲畜养殖者造成经济困难。 3D8单链可变片段(scFv)蛋白是具有核酸酶活性的重组抗体,对小鼠和鸡的各种DNA和RNA病毒均表现出抗病毒作用。在该实验中,通过基因组PCR和免疫组化分析筛选了通过将3D8 scFv G转基因公鸡与野生母鸡杂交而产生的3D8 scFv G转基因鸡。 3D8 scFv转基因鸡,野生型同胞鸡和SPF鸡被IBV直接感染(每组5只鸡),并通过空中传播间接感染(每组15只鸡)。在眼内感染后第3天和第5天,使用口咽和泄殖腔拭子通过定量实时PCR测量相对IBV滴度。接触传播组的3D8 scFv转基因鸡的病毒载量显着降低。另外,在感染后第17天从每个组收集血液。 ELISA结果显示,来自接触传播组的3D8 scFv转基因鸡中针对IBV的抗体效价显着降低。这些结果表明3D8 scFv蛋白潜在地抑制了鸡中传染性支气管炎病毒的传播。

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