Design and validation of VLP/mRNA-based vaccine strategies against SARS-CoV2

NIH Pandemic-Era Grants

Pandemic Era Grants

2021

↗ Live URL (may be dead)

Document text

Principal Investigator: paolo  lusso
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2021
Award: $121,121
Funding agency: National Institute of Allergy and Infectious Diseases

The project has yielded the following results:
1.	Synthesis of tail-modified SARS-CoV2 envelope spike (S) mRNAs to facilitate VLP formation with lentivirus core proteins. In order to facilitate the assembly of VLPs containing lentivirus (HIV-1 or SIV) core proteins, we have designed S proteins modified in their cytoplasmic tails (CT) by replacement of the natural tail with either the HIV-1 or SIV gp41 CT.  A truncated form of the lentivirus tails has been used, based on our original design for the HIV-1 vaccine, to promote a more efficient surface membrane expression.  Moderna Inc. has produced the specific mRNAs according to our design.
2.	Recombinant expression of the SARS-CoV2 envelope spike (S) glycoprotein in mammalian cells. The mRNA expressing the various CoV2 envelope spike (S) glycoproteins (native, HIV-CT and SIV-CT chimeric) produced by Moderna Inc. were tested in mammalian cells to verify their expression both in terms of efficiency and antigenic profile using commercially available monoclonal antibodies (mAbs) against the S protein.
3.	Production of VLPs. The spike protein mRNAs were co-transfected into mammalian cells in all possible combinations with mRNAs encoding HIV-1 or SIV Gag or Gag-Pol. The extracellular VLPs were harvested and concentrated by ultracentrifugation on sucrose cushions.
4.	Extensive characterization of VLPs. The produced VLPs have been extensively characterized both quantitatively and qualitatively using a wide array of physical and immunological assays, including virion capture by mAbs followed by ELISA for lentivirus core antigen quantification, Western blot and others.

Our results indicate that VLPs comprising the CoV2 Spike protein and a lentiretrovirus core can be efficiently produced and display the native-like form of the S-protein on their surface. Preliminary results from immunogenicity studies in a small animal model (mice) indicate that VLP-based mRNA vaccine approaches are superior to single-protein mRNA vaccines.

Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV vaccine><AIDS Virus><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Animal Model><Animal Models and Related Studies><Antigens><COVID-19 vaccine><COVID-19 virus><COVID19 vaccine><COVID19 virus><Clinical Research><Clinical Study><Clinical Treatment Moab><CoV emergence><CoV-2><CoV2><Core Protein><Cytoplasmic Domain><Cytoplasmic Tail><Development><ELISA><Enzyme-Linked Immunosorbent Assay><Glycoproteins><HIV><HIV-1><HIV-1 vaccine><HIV-I><HIV1><HIV1 vaccine><Harvest><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human immunodeficiency virus 1><Immune response><Immunological response><Immunology procedure><Individual><LAV-HTLV-III><Lentivirinae><Lentivirus><Lymphadenopathy-Associated Virus><Mammalian Cell><Membrane><Messenger RNA><Mice><Mice Mammals><Monoclonal Antibodies><Murine><Mus><Phase><Production><Proteins><RNA vaccine><RNA-based vaccine><Recombinants><SARS corona virus 2><SARS-CoV-2><SARS-CoV-2 vaccine><SARS-CoV2><SARS-CoV2 vaccine><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-coronavirus-2 vaccine><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><SIV><SIV envelope protein gp41><SIV gp40><SIV gp41><Saccharose><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV 2 vaccine><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 vaccine><Severe acute respiratory syndrome related corona virus 2><Simian Immunodeficiency Viruses><Sucrose><Surface><Tail><Testing><Ultracentrifugation><Vaccination><Vaccines><Validation><Virion><Virus><Virus Assembly><Virus Particle><Virus-HIV><Virus-Lenti><Virus-like particle><Western Blotting><Western Immunoblotting><Wuhan coronavirus><base><corona virus disease 2019 vaccine><corona virus emergence><coronavirus disease 2019 vaccine><coronavirus disease 2019 virus><coronavirus emergence><design><designing><developmental><emergent CoV><emergent corona virus><emergent coronavirus><emerging CoV><emerging corona virus><emerging coronavirus><experience><extracellular><hCoV19><host response><immune system response><immunogen><immunogenicity><immunologic assay><immunologic assay/test><immunoresponse><mAbs><mRNA><mRNA vaccine><mRNA-based vaccine><membrane structure><model of animal><model organism><nCoV><nCoV2><new CoV><new corona virus><new coronavirus><novel CoV><novel corona virus><novel coronavirus><protein blotting><vaccine against 2019-nCov><vaccine against SARS-CoV-2><vaccine against SARS-CoV2><vaccine against SARS-coronavirus-2><vaccine against Severe Acute Respiratory Syndrome CoV 2><vaccine against Severe acute respiratory syndrome coronavirus 2><vaccine for novel coronavirus><viral assembly><virus-like nanoparticles><viruslike particle>