Characterization of an H4N2 influenza virus from Quails with a multibasic motif in the hemagglutinin cleavage site

•An H4N2 influenza virus with a multibasic cleavage site in the hemagglutinin protein was isolated from quails.•This virus remained lowly pathogenic in chickens and required trypsin for in vitro growth.•This virus showed higher preference for mammalian-type sialic acid receptors.•This virus transmitted better in chicken than a duck-origin H4N2 virus.

The cleavage motif in the hemagglutinin (HA) protein of highly pathogenic H5 and H7 subtypes of avian influenza viruses is characterized by a peptide insertion or a multibasic cleavage site (MBCS). Here, we isolated an H4N2 virus from quails (Quail/CA12) with two additional arginines in the HA cleavage site, PEKRRTR/G, forming an MBCS-like motif. Quail/CA12 is a reassortant virus with the HA and neuraminidase (NA) gene most similar to a duck-isolated H4N2 virus, PD/CA06 with a monobasic HA cleavage site. Quail/CA12 required exogenous trypsin for efficient growth in culture and caused no clinical illness in infected chickens. Quail/CA12 had high binding preference for α2,6-linked sialic acids and showed higher replication and transmission ability in chickens and quails than PD/CA06. Although the H4N2 virus remained low pathogenic, these data suggests that the acquisition of MBCS in the field is not restricted to H5 or H7 subtypes.