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📄 原文题目

Recombinant influenza A H5N1 viruses expressing fluorescent and luminescent reporter proteins

🔗 原文链接

💡 AI 核心解读

开发了基于H5N1病毒的新型报告系统,通过在NS基因中插入荧光/发光蛋白(NLuc、miniGFP2、UnaG)实现病毒复制的实时监测,同时验证了其在体外复制效率、体内致病性及血清学检测中的应用价值。

📝 英文原版摘要

Highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.4.4b viruses present a broad host range, with recent spillover and sustained transmission in dairy cattle in the United States. Replication-competent reporter viruses are critical tools that enable real-time monitoring of virus replication facilitating high-throughput antiviral and serological screens. In this study, we engineered three recombinant H5N1 clade 2.3.4.4b reporter viruses expressing nanoluciferase (NLuc) and two fluorescent reporter proteins, miniGFP2 and UnaG within the open reading frame of the nonstructural (NS) gene of the bovine A/Cattle/Texas/063224-24-1/2024 (TX2/24) virus. All reporter viruses replicated efficiently in vitro, presenting replication kinetics comparable to the parental rTX2/24 virus, but exhibited smaller plaque sizes suggesting reduced cell-to-cell spread. In vivo infection studies in mice showed comparable pathogenicity among all four viruses, although rTX2/24-miniGFP2 and rTX2/24-UnaG exhibited decreased virus shedding relative to rTX2/24 and rTX2/24-NLuc. Virus titrations and in situ localization of virus replication sites demonstrated robust replication in respiratory tissues, with slightly attenuated systemic dissemination of all three reporter viruses. Fluorescent virus neutralization assays using miniGFP2 and UnaG reporter viruses accurately quantified neutralizing antibody titers in sera from naturally infected dairy cattle, consistent with wild-type virus assays. Collectively, these results establish the TX2/24-based reporter viruses as versatile and biologically relevant tools for investigating H5N1 pathogenesis, and serological screens and potentially antiviral screens.
工程化赛博基因细胞疗法牛源和人源甲型H5N1流感病毒在小鼠和仓鼠模型中的感染与传播动力学
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