A single N342D substitution in Influenza B Virus NA protein determines viral pathogenicity in mice

Influenza B virus (IBV) is one of the most important human respiratory viruses: it causes approximately one-third of the global influenza-related disease burden each year. However, compared with the several pathogenicity-related molecular markers that have been identified for influenza A virus (IAV), little is known about potential IBV pathogenicity-related markers. Here, although the IBV strain B/Anhui-Tunxi/1528/2014 (AH1528/14) exhibited a more efficient replication ability <i>in vitro</i> and higher pathogenicity <i>in vivo</i> compared with IBV strain B/Anhui-Baohe/127/2015 (AH127/15), only three amino acids differences (HA_A390E, NA_N342D and PB1_V212I) were observed among their full genomes. The contributions of each amino acid difference to the virus pathogenicity were further investigated. Compared with the wild type IBV virus rAH127, the recombinant virus harbouring a single substitution of HA_A390E had a similar phenotype, whereas the recombinant virus harbouring PB1_V212I replicated to a moderately higher titre in both MDCK cells and in mice. Notably, the virus harbouring NA_N342D showed significantly better growth properties in MDCK cells and higher fatality rates in mice. In addition, the presence of NA_N342D dramatically enhanced the viral neuraminidase activity. In conclusion, our study identified a novel IBV molecular marker, NA_N342D, that could significantly increase the virulence of IBV in mice.