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Principal Investigator: Wing Pui Kong
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2024
Award: $912,113
Funding agency: National Institute of Allergy and Infectious Diseases
Human paramyxoviruses and pneumoviruses are widespread pathogens, cause considerable disease burden. Parainfluenza virus types 1-4 (PIV1-4) are highly infectious human pathogens, of which PIV3 is most commonly responsible for severe respiratory illness in newborns, elderly, and immunocompromised individuals. Based on a cryoelectron microscopy structure of an engineered PIV3 F prefusion-stabilized trimer, we engineered in additional mutations different from our previous published modifications in each of the four PIV fusion (F) glycoproteins to further stabilize their metastable prefusion states. The vaccine candidates were able to elicit murine PIV type-specific response, with little cross-neutralization of other PIVs. We evaluated the neutralization responses of the vaccine candidates in nonhuman primates (NHPs). We applied similar concept and designs to hMPV F proteins. We designed various mutations to further stabilize the PIV F prefusion trimers and hMPV prefusion trimers. We investigated the impact of further stabilizing HMPV F in the pre-F state. We replaced the furin-cleavage site with a flexible linker, creating a single chain F that yielded increased amounts of pre-F stabilized trimers, enabling the generation and assessment of F trimers stabilized by multiple disulfide bonds. Immunogenicity assessments in mice showed the triple disulfide-stabilized pre-F trimer could elicit high titer neutralization than elicited by post-F. Immunogenicity assessments in pre-exposed rhesus macaques showed the triple disulfide-stabilized pre-F could recall high neutralizing titers after a single immunization, with little discrimination in the recall response between pre-F and post-F. Collectively, these results suggest single-chain triple disulfide-stabilized pre-F trimers to be promising HMPV-vaccine antigens. In the past year, we designed the modified RSV and HMPV stabilized trimers on the surface of self-assemble encapsulins nanoparticles (EN) as vaccine immunogens. Mice immunogenicity assessments of the mosaic RSV and HMPV F trimers EN nanoparticles immunogens indicated significant improved immune responses against HMPV trimers but with similar response to RSV trimmer even both immunogens were on the same nanoparticles. Our further design modifications of the trimers are on-going and animal studies will follow. For PIV vaccine development, we did proteins immunization of the novel stabilized F trimmers of PIV2 and they induced high neutralization titers against PIV2 live viruses.
Terms: <0-4 weeks old><Animals><Antigens><Chimera Protein><Chimeric Proteins><Cognitive Discrimination><Cryo-electron Microscopy><Cryoelectron Microscopy><Discrimination><Disulfides><Elderly><Electron Cryomicroscopy><Engineering><Fusion Protein><Generations><Genetic Alteration><Genetic Change><Genetic defect><Glycoproteins><HPIV1><Hemadsorption Type 2 Virus><Hemadsorption Virus 2><Human><Human Parainfluenza Virus 1><Immune response><Immunization><Immunocompromised><Immunocompromised Host><Immunocompromised Patient><Immunological response><Immunosuppressed Host><Individual><Infection><M mulatta><M. mulatta><Macaca mulatta><Mice><Mice Mammals><Modern Man><Modification><Murine><Mus><Mutation><Newborn Infant><Newborns><Para-Influenza Virus Type 1><Paramyxoviridae><Paramyxovirus><Pneumoviridae><Pneumovirinae><Pneumovirus><Proteins><Publishing><Rhesus><Rhesus Macaque><Rhesus Monkey><Site><Structure><Surface><Vaccine Antigen><Vaccines><Virus><advanced age><burden of disease><burden of illness><cryo-EM><cryoEM><cryogenic electron microscopy><design><designing><develop a vaccine><develop vaccines><development of a vaccine><disease burden><disulfide bond><flexibility><flexible><genome mutation><geriatric><host response><human pathogen><immune response to vaccination><immune response to vaccines><immune system response><immunogen><immunogenicity><immunoresponse><immunosuppressed patient><improved><mosaic><nano particle><nano-sized particle><nanoparticle><nanosized particle><newborn child><newborn children><non-human primate><nonhuman primate><novel><parainfluenza virus type 1><pathogen><pre-clinical><preclinical><respiratory><respiratory virus><response><self assembly><senior citizen><vaccine associated immune response><vaccine candidate><vaccine development><vaccine immune response><vaccine immunogenicity><vaccine induced immune response><vaccine response><vaccine responsiveness><vaccine-induced response>