Hubby Bolyn, Talarico Todd, Maughan Maureen, Reap Elizabeth A, Berglund Peter, Kamrud Kurt I, Copp Laura, Lewis Whitney, Cecil Chad, Norberg Pamela, Wagner Jordan, Watson Aubrey, Negri Sarah, Burnett Bruce K, Graham Andrew, Smith Jonathan F, Chulay Jeffrey D
AlphaVax, Inc., 2 Triangle Drive, Research Triangle Park, NC 27709, USA.
Vaccine. 2007 Nov 23;25(48):8180-9. doi: 10.1016/j.vaccine.2007.09.038. Epub 2007 Oct 5.
We used a propagation-defective, single-cycle, alphavirus replicon vector system to produce virus-like replicon particles (VRP) expressing the hemagglutinin (HA) and neuraminidase (NA) proteins from influenza A/Wyoming/03/2003 (H3N2). Efficient production methods were scaled to produce pilot lots of HA VRP and NA VRP and clinical lots of HA VRP. HA VRP-induced high-titered antibody responses in mice, rabbits and rhesus macaques, as measured by ELISA or hemagglutination inhibition (HI) assays, and robust cellular immune responses in mice and rhesus macaques, as measured by IFN-gamma ELISPOT. NA VRP also induced cellular immune responses in mice. A toxicology study with HA VRP and NA VRP in rabbits showed no adverse effects in any parameter. These studies support clinical testing of alphavirus replicon vaccines for influenza.
我们使用了一种复制缺陷型、单周期的甲病毒复制子载体系统来生产表达来自甲型流感病毒/怀俄明州/03/2003(H3N2)的血凝素(HA)和神经氨酸酶(NA)蛋白的病毒样复制子颗粒(VRP)。高效的生产方法经扩大规模后用于生产HA VRP和NA VRP的中试批次以及HA VRP的临床批次。通过ELISA或血凝抑制(HI)试验测定,HA VRP在小鼠、兔子和恒河猴中诱导了高滴度抗体反应,通过IFN-γ ELISPOT测定,在小鼠和恒河猴中诱导了强烈的细胞免疫反应。NA VRP在小鼠中也诱导了细胞免疫反应。一项对兔子进行的HA VRP和NA VRP毒理学研究显示,任何参数均未出现不良反应。这些研究支持对甲型流感病毒复制子疫苗进行临床试验。