Seasonal influenza (flu) virus, an NIAID category C priority pathogen, causes widespread infection, resulting in at least 3-5 million cases of severe illness and 250,000-500,000 deaths worldwide. While annual vaccination is recommended for all individuals aged over 6 months in the US, current vaccines are only around 60% effective and there is an urgent need for safe, more effective influenza vaccines that offer broad-spectrum protection. To meet this need, FluGen has developed a novel, live replication-deficient influenza virus, M2SR, that can be modified to encode the viral antigens from any influenza A or B strain. We have shown that the influenza A strains of M2SR (M2SR) provide effective protection against both homologous and heterologous viruses in both mice and ferrets. In our previous Phase I award, we generated an influenza B M2SR (BM2SR) vaccine and demonstrated that it also provided effective protection against homologous and heterologous influenza B viruses in a mouse model. Interestingly, there appear to be some differences in the mechanism of protection between M2SR and BM2SR vaccines. In the current Phase II grant, we will investigate the efficacy and safety of the BM2SR vaccine in a ferret model, which more closely resembles the viral infection in humans. We hypothesize that the vaccine will provide safe and effective protection against both homologous and heterologous influenza B infection in ferrets. We will also investigate the mechanism of protection in greater depth. Aim 1. To determine the efficacy and spectrum of protection afforded by the vaccine in ferrets. Aim 2. To evaluate the safety of the vaccine in ferrets. Aim 3. To investigate the mechanism of protection induced by BM2SR. These studies will provide a comprehensive pre-clinical evaluation of the efficacy and safety of the BM2SR vaccine. The outcome of the studies is expected to have high impact on the fundamental understanding of the mechanism of protection of the BM2SR influenza B vaccine and to progress FluGen?s vaccine program toward a quadrivalent human clinical trial.