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Powassan virus (POWV) is an emerging tick borne flavivirus (TBFV) that causes severe neuroinvasive disease. Currently, there are no approved treatments or vaccines to combat POWV infection. Here, we generated and characterized a nanoparticle immunogen displaying domain III (EDIII) of the POWV E glycoprotein. Immunization with POWV EDIII presented on nanoparticles resulted in significantly higher serum neutralizing titers against POWV than immunization with monomeric POWV EDIII. Furthermore, passive transfer of EDIII-reactive sera protected against POWV challenge in vivo. We isolated and characterized a panel of EDIII-specific monoclonal antibodies (mAbs) and identified several that potently inhibit POWV infection and engage distinct epitopes within the lateral ridge and C-C′ loop of the EDIII. By creating a subunit-based nanoparticle immunogen with vaccine potential that elicits antibodies with protective activity against POWV infection, our findings enhance our understanding of the molecular determinants of antibody-mediated neutralization of TBFVs. Author summary: Powassan virus (POWV) is an emerging tick-borne flavivirus that circulates in North America. Infection can cause severe neuro-invasive disease, and currently there is no treatment or vaccine against POWV. We generated a novel POWV immunogen that displays the EDIII of the envelope glycoprotein on the surface of a self-assembling protein nanoparticle. We found that immunization of the EDIII-displaying nanoparticle in mice elicited potently neutralizing and protective antibodies. We further characterized EDIII-specific neutralizing mAbs and mapped their epitopes to specific regions within the POWV EDIII. Our study sheds light on determinants of antibody-mediated neutralization and protection against POWV and other TBFVs. It also informs EDIII-based vaccine and antiviral efforts to combat POWV infection. [ABSTRACT FROM AUTHOR]

The mosquito-borne West Nile virus (WNV) causes human and animal disease with outbreaks in several parts of the world including North America, the Mediterranean countries, Central and East Europe, the Middle East, and Africa. Particularly in elderly people and individuals with an impaired immune system, infection with WNV can progress into a serious neuroinvasive disease. Currently, no treatment or vaccine is available to protect humans against infection or disease. The goal of this study was to develop a WNV-vaccine that is safe to use in these high-risk human target populations. We performed a vaccine efficacy study in non-human primates using the contemporary, pathogenic European WNV genotype 1a challenge strain, WNV-Ita09. Two vaccine strategies were evaluated in rhesus macaques (Macaca mulatta) using recombinant soluble WNV envelope (E) ectodomain adjuvanted with Matrix-M, either with or without DNA priming. The DNA priming immunization was performed with WNV-DermaVir nanoparticles. Both vaccination strategies successfully induced humoral and cellular immune responses that completely protected the macaques against the development of viremia. In addition, the vaccine was well tolerated by all animals. Overall, The WNV E protein adjuvanted with Matrix-M is a promising vaccine candidate for a non-infectious WNV vaccine for use in humans, including at-risk populations. [ABSTRACT FROM AUTHOR]

West Nile virus (WNV) is a mosquito-borne flavivirus that infects humans and other mammals. In some cases WNV causes severe neurological disease. During recent years, outbreaks of WNV are increasing in worldwide distribution and novel genetic variants of the virus have been detected. Although a substantial amount of data exists on WNV infections in rodent models, little is known about early events during WNV infection in primates, including humans. To gain a deeper understanding of this process, we performed experimental infections of rhesus macaques and common marmosets with a virulent European WNV strain (WNV-Ita09) and monitored virological, hematological, and biochemical parameters. WNV-Ita09 productively infected both monkey species, with higher replication and wider tissue distribution in common marmosets compared to rhesus macaques. The animals in this study however, did not develop clinical signs of WNV disease, nor showed substantial deviations in clinical laboratory parameters. In both species, the virus induced a rapid CD56dimCD16bright natural killer response, followed by IgM and IgG antibody responses. The results of this study show that healthy rhesus macaques and common marmosets are promising animal models to study WNV-Ita09 infection. Both models may be particularly of use to evaluate potential vaccine candidates or to investigate WNV pathogenesis. Author Summary: West Nile virus (WNV) is a mosquito-borne virus that can infect mammals, including humans. Most infected humans do not develop disease, but in about 20% of cases humans develop WNV-related disease symptoms, varying in severity from fever to a sometimes life-threatening neuro-invasive disease. The number of WNV infections in Europe has increased in recent years and is caused by viruses that are genetically different from the viruses that caused the WNV epidemic in North America. In this study, we have experimentally infected two different monkey species, rhesus macaques and common marmosets, with the European WNV isolate Ita09 to evaluate the early events after infection and the onset of the disease. Both species were equally susceptible to infection with WNV-Ita09, but differences between species were observed. Compared to rhesus macaques, common marmosets had higher virus loads in blood, and presented a wider distribution of the virus in various organs. Based on the analysis of virological, immunological, biochemical and hematological parameters, we conclude that rhesus macaques as well as common marmosets are potentially useful animal models to evaluate vaccine candidates or to investigate WNV pathogenesis. [ABSTRACT FROM AUTHOR]

Eastern equine encephalitis virus (EEEV) is one of the most virulent viruses endemic to North America. No licensed vaccines or antiviral therapeutics are available to combat this infection, which has recently shown an increase in human cases. Here, we characterize human monoclonal antibodies (mAbs) isolated from a survivor of natural EEEV infection with potent (<20 pM) inhibitory activity of EEEV. Cryo-electron microscopy reconstructions of two highly neutralizing mAbs, EEEV-33 and EEEV-143, were solved in complex with chimeric Sindbis/EEEV virions to 7.2 Å and 8.3 Å, respectively. The mAbs recognize two distinct antigenic sites that are critical for inhibiting viral entry into cells. EEEV-33 and EEEV-143 protect against disease following stringent lethal aerosol challenge of mice with highly pathogenic EEEV. These studies provide insight into the molecular basis for the neutralizing human antibody response against EEEV and can facilitate development of vaccines and candidate antibody therapeutics. • Potent human anti-EEEV monoclonal antibodies were isolated from an EEEV survivor • Fabs neutralized virus, but bivalent IgG was optimal for neutralization • Antibodies inhibit virus entry by stabilization of virus particles • EEEV-33 (IgG) and EEEV-143 (IgA) protected against aerosolized EEEV challenge Eastern equine encephalitis virus (EEEV) causes rare but serious and often fatal infection associated with inflammation of the brain. Monoclonal antibodies identified from a human survivor recognize distinct epitopes to prevent viral entry recognize, demonstrate protection as well as potent neutralization against EEEV, and may inform future vaccine design. [ABSTRACT FROM AUTHOR]

West Nile virus (WNV) is a neurotropic flavivirus that is now a primary cause of epidemic encephalitis in North America. Studies of mice have demonstrated that the humoral immune response against WNV limits primary infection and protects against a secondary challenge. The most-potent neutralizing mouse monoclonal antibodies (MAbs) recognize an epitope on the lateral ridge of domain III (DIII-lr) of the envelope (E) protein. However, studies with serum from human patients show that antibodies against the DIII-lr epitope comprise, at best, a minor component of the human anti-WNV antibody response. Herein, we characterize in detail two WNV-specific human MAbs, CR4348 and CR4354, that were isolated from B-cell populations of convalescent patients. These MAbs strongly neutralize WNV infection of cultured cells, protect mice against lethal infection in vivo, and yet poorly recognize recombinant forms of the E protein. Instead, CR4348 and CR4354 bind determinants on intact WNV virions and subviral particles in a pH-sensitive manner, and neutralization is altered by mutations at the dimer interface in domain II and the hinge between domains I and II, respectively. CR4348 and CR4354 human MAbs neutralize infection at a postattachment step in the viral life cycle, likely by inhibiting acid-induced fusion within the endosome. [ABSTRACT FROM AUTHOR]