1. Targeting intra-viral conserved nucleocapsid (N) proteins as novel vaccines against SARS-CoVs
- Author
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Qi Zeng, Cheng William Hong, Min Thura, Jie Li, Jimmy Hong, Abhishek Gupta, Joel Sng, and Koon Hwee Ang
- Subjects
Immunogen ,COVID-19 Vaccines ,viruses ,Biophysics ,Biology ,medicine.disease_cause ,Antibodies, Viral ,Biochemistry ,Virus ,Epitope ,Antibodies, Monoclonal, Murine-Derived ,Epitopes ,Mice ,Immune system ,Immunogenicity, Vaccine ,Immunity ,Virology ,medicine ,Animals ,Coronavirus Nucleocapsid Proteins ,Humans ,Molecular Biology ,Pandemics ,Research Articles ,Coronavirus ,Immune Evasion ,Nucleocapsid protein ,Sequence Homology, Amino Acid ,SARS-CoV-2 ,Pharmacology & Toxicology ,COVID-19 ,Cell Biology ,Th1 Cells ,Vaccination ,Severe acute respiratory syndrome-related coronavirus ,Models, Animal ,Spike Glycoprotein, Coronavirus ,Vaccines, Subunit ,biology.protein ,Antibody ,Vaccine - Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the global pandemic of the Coronavirus disease in late 2019 (COVID-19). Vaccine development efforts have predominantly been aimed at 'Extra-viral' Spike (S) protein as vaccine vehicles, but there are concerns regarding ‘viral immune escape’ since multiple mutations may enable the mutated virus strains to escape from immunity against S protein. The ‘Intra-viral’ Nucleocapsid (N-protein) is relatively conserved among mutant strains of coronaviruses during spread and evolution. Herein, we demonstrate novel vaccine candidates against SARS-CoV-2 by using the whole conserved N-protein or its fragment/peptides. Using ELISA assay, we showed that high titers of specific anti-N antibodies (IgG, IgG1, IgG2a, IgM) were maintained for a reasonably long duration (> 5 months), suggesting that N-protein is an excellent immunogen to stimulate host immune system and robust B-cell activation. We synthesized three peptides located at the conserved regions of N-protein among CoVs. One peptide showed as a good immunogen for vaccination as well. Cytokine arrays on post-vaccination mouse sera showed progressive up-regulation of various cytokines such as IFN-γ and CCL5, suggesting that TH1 associated responses are also stimulated. Furthermore, vaccinated mice exhibited an elevated memory T cells population. Here, we propose an unconventional vaccine strategy targeting the conserved N-protein as an alternative vaccine target for coronaviruses. Moreover, we generated a mouse monoclonal antibody specifically against an epitope shared between SARS-CoV and SARS-CoV-2, and we are currently developing the First-in-Class humanized anti-N-protein antibody to potentially treat patients infected by various CoVs in the future.
- Published
- 2021