Designing multivalent immunogens for alphavirus vaccine optimization

There is a pressing need for vaccines against mosquito-borne alphaviruses such as Venezualen and eastern equine encephalitis viruses (VEEV, EEEV). We demonstrate an approach to vaccine development based on physicochemical properties (PCP) of amino acids to design a PCP-consensus sequence of the epitope-rich B domain of the VEEV major antigenic E2 protein. The consensus “spike” domain was incorporated into a live-attenuated VEEV vaccine candidate (ZPC/IRESv1). Mice inoculated with either ZPC/IRESv1 or the same virus containing the consensus E2 protein fragment (VEEVconE2) were protected against lethal challenge with VEEV strains ZPC-738 and 3908, and Mucambo virus (MUCV, related to VEEV), and had comparable neutralizing antibody titers against each virus. Both vaccines induced partial protection against Madariaga virus (MADV), a close relative of EEEV, lowering mortality from 60% to 20%. Thus PCP-consensus sequences can be integrated into a replicating virus that could, with further optimization, provide a broad-spectrum vaccine against encephalitic alphaviruses. •Novel consensus vaccine (VEEVconE2) based on physicochemical properties (PCP) of amino acids.•The ZPC/IRESv1 vaccine containing the VEEVconE2 spike was stable and immunogenic.•Both ZPC/IRESv1 and the VEEVconE2 protect mice from lethal challenge with 3 wild-type VEEV complex strains.•Both vaccines provided partial protection against challenge from Madariaga Virus of the EEE complex.