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Principal Investigator: Barney Graham
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2019
Award: $951,522
Funding agency: National Institute of Allergy and Infectious Diseases
When the MERS CoV outbreak raised global health concerns, we initiated a program to develop a candidate vaccine. Starting from Spike glycoprotein (S) sequences, we developed an immunization strategy consisting of a full-length S DNA prime and a S1 subunit protein boost that elicited high titers of neutralizing antibodies against eight different MERS-CoV strains. Immune sera contained potent neutralizing antibodies targeting the receptor binding domain (RBD), non-RBD portions of S1, and the S2 subunit. From the immunized mice, we produced a panel of hybridomas and produced monoclonal antibodies from which a variety with high neutralizing activity were selected for further characterization. The atomic structure of a monoclonal antibody, D12, in complex with the RBD revealed two distinct mechanisms by which they block binding to the MERS-CoV receptor, DPP4. The full-length S protein was also immunogenic in non-human primates. These initial studies demonstrated that coronovirus spike vaccine immunogens induced a diverse repertoire of neutralizing antibodies, demonstrating an efficient approach to vaccine design that may be applicable to other emerging viruses. Through collaborations, we have solved the structure of the MERS S protein and based on this structure, designed stabilizing mutations which stabilize the S protein in its prefusion conformation. We are currently assessing immunogenicity of the stabilized MERS S protein, and evaluating stabilizing mutations in other coronavirus spike proteins. Additionally, we have continued to characterize the binding and functional characteristics of monoclonal antibodies that are specific for different epitopes on the S protein.
Terms: <ATGN><Antigenic Determinants><Antigens><Antisera><Binding><Binding Determinants><Characteristics><Clinical Treatment Moab><Collaborations><Complex><Coronaviridae><Coronavirus><Coronavirus spike protein><DNA><Deoxyribonucleic Acid><Development><Disease Outbreaks><Epitopes><General Viruses><Genetic Alteration><Genetic Change><Genetic defect><Glycoproteins><Goals><Hybridomas><Immune Sera><Immunization><Immunize><Immunologic Sensitization><Immunologic Stimulation><Immunological Sensitization><Immunological Stimulation><Immunostimulation><Length><MERS-CoV><Mice><Mice Mammals><Middle East Respiratory Syndrome><Middle East Respiratory Syndrome Coronavirus><Molecular Configuration><Molecular Conformation><Molecular Interaction><Molecular Stereochemistry><Monoclonal Antibodies><Murine><Mus><Mutation><Outbreaks><Protein Subunits><Proteins><Public Health><Receptor Protein><SARS Virus><SARS coronavirus><SARS-Associated Coronavirus><SARS-CoV><SARS-Related Coronavirus><Severe Acute Respiratory Syndrome Virus><Structure><Technology><Urbani SARS-Associated Coronavirus><Vaccine Design><Vaccines><Virus><base><conformation><conformational state><design><designing><develop a vaccine><development of a vaccine><developmental><genome mutation><global health><immune serum><immunogen><immunogenic><immunogenicity><insight><mAbs><neutralizing antibody><non-human primate><nonhuman primate><pandemic><pandemic disease><programs><receptor><receptor binding><receptor bound><vaccine candidate><vaccine development><vaccine formulation>