Protective antibodies against Eastern equine encephalitis virus bind to epitopes in domains A and B of the E2 glycoprotein

Eastern equine encephalitis virus (EEEV) is a mosquito-transmitted alphavirus with a high case mortality rate in humans. EEEV is a biodefense concern because of its potential for aerosol spread and the lack of existing countermeasures. In this study, we identified a panel of 18 neutralizing murine monoclonal antibodies (mAbs) against the EEEV E2 protein, several of which had “elite” activity with 50% and 99% inhibitory concentrations (EC(50) and EC(99)) of less than 10 and 100 ng/ml, respectively. Alanine-scanning mutagenesis and neutralization escape mapping analysis revealed epitopes for these mAbs in domains A or B of the E2 glycoprotein. A majority of the neutralizing mAbs blocked at a post-attachment stage, with several inhibiting viral membrane fusion. Administration of one dose of anti-EEEV mAbs protected mice from lethal subcutaneous or aerosol challenge. These experiments define the mechanistic basis for neutralization by protective anti-EEEV mAbs and suggest a path forward for treatment and vaccine design.