Center for Systems Medicine, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China; Suzhou Institute of Systems Medicine, Suzhou 215123, China; Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Cell Host Microbe. 2017 Mar 8;21(3):334-343. doi: 10.1016/j.chom.2017.02.007.
New influenza vaccines that provide effective and broad protection are desperately needed. Live attenuated viruses are attractive vaccine candidates because they can elicit both humoral and cellular immune responses. However, recent formulations of live attenuated influenza vaccines (LAIVs) have not been protective. We combined high-coverage transposon mutagenesis of influenza virus with a rapid high-throughput screening for attenuation to generate W7-791, a live attenuated mutant virus strain. W7-791 produced only a transient asymptomatic infection in adult and neonatal mice even at doses 100-fold higher than the LD of the parent strain. A single administration of W7-791 conferred full protection to mice against lethal challenge with H1N1, H3N2, and H5N1 strains, and improved viral clearance in ferrets. Adoptive transfer of T cells from W7-791-immunized mice conferred heterologous protection, indicating a role for T cell-mediated immunity. These studies present an LAIV development strategy to rapidly generate and screen entire libraries of viral clones.
急需提供有效且广泛保护的新型流感疫苗。活减毒病毒是有吸引力的疫苗候选物，因为它们可以引发体液和细胞免疫反应。然而，最近的活减毒流感疫苗（LAIV）配方并没有起到保护作用。我们将流感病毒的高覆盖率转座子诱变与快速高通量筛选相结合，以产生 W7-791，这是一种活减毒突变病毒株。W7-791 甚至在比亲本株 LD 高 100 倍的剂量下，也仅在成年和新生小鼠中引起短暂的无症状感染。单次给予 W7-791 可使小鼠完全免受 H1N1、H3N2 和 H5N1 株的致死性攻击，并改善雪貂中的病毒清除。从 W7-791 免疫小鼠中过继转移 T 细胞可赋予异源保护，表明 T 细胞介导的免疫起作用。这些研究提出了一种 LAIV 开发策略，可以快速生成和筛选整个病毒克隆文库。
