Li Junwei, Arévalo Maria T, Diaz-Arévalo Diana, Chen Yanping, Choi Jang-Gi, Zeng Mingtao
Center of Excellence for Infectious Diseases, Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, 5001 El Paso Drive, El Paso, TX 79905, USA.
Center of Excellence for Infectious Diseases, Department of Biomedical Sciences, Paul L. Foster School of Medicine, Texas Tech University Health Sciences Center, 5001 El Paso Drive, El Paso, TX 79905, USA.
J Control Release. 2015 Jun 10;207:70-6. doi: 10.1016/j.jconrel.2015.04.001. Epub 2015 Apr 7.
Vaccination with live attenuated vaccines (LAVs) is an effective way for prevention of infectious disease. While several methods are employed to create them, efficacy and safety are still a challenge. In this study, we evaluated the feasibility of creating a self-attenuated RNA virus expressing a functional species-specific artificial microRNA. Using influenza virus as a model, we produced an attenuated virus carrying a mammalian-specific miR-93 expression cassette that expresses a viral nucleoprotein (NP)-specific artificial microRNA from an insertion site within the non-structural (NS) gene segment. The resulting engineered live-attenuated influenza virus, PR8-amiR-93NP, produced mature and functional artificial microRNA against NP in mammalian cells, but not in avian cells. Furthermore, PR8-amiR-93NP was attenuated by 10(4) fold in mice compared with its wild-type counterpart. Importantly, intranasal immunization with PR8-amiR-93NP conferred cross-protective immunity against heterologous influenza virus strains. In short, this method provides a safe and effective platform for creation of live attenuated RNA viral vaccines.
接种减毒活疫苗（LAVs）是预防传染病的有效方法。虽然有几种方法可用于制备减毒活疫苗，但其有效性和安全性仍是一个挑战。在本研究中，我们评估了构建一种表达功能性物种特异性人工微小RNA的自我减毒RNA病毒的可行性。以流感病毒为模型，我们构建了一种携带哺乳动物特异性miR-93表达盒的减毒病毒，该表达盒从非结构（NS）基因片段内的插入位点表达病毒核蛋白（NP）特异性人工微小RNA。产生的工程化减毒活流感病毒PR8-amiR-93NP在哺乳动物细胞中产生了针对NP的成熟且功能性的人工微小RNA，但在禽细胞中未产生。此外，与野生型相比，PR8-amiR-93NP在小鼠中的毒力降低了10^4倍。重要的是，用PR8-amiR-93NP进行鼻内免疫可赋予针对异源流感病毒株的交叉保护性免疫。简而言之，该方法为构建减毒活RNA病毒疫苗提供了一个安全有效的平台。
