Document text
Principal Investigator: JAY A BERZOFSKY
Organization: DIVISION OF BASIC SCIENCES - NCI
Fiscal Year: 2023
Award: $2,147,209
Funding agency: National Cancer Institute
In vivo vaccine studies in macaques and hamsters: Rhesus macaques have been primed IM with S1 spike protein in different adjuvants and boosted systemically with spike in alum or mucosally intranasally with spike in nanoparticles with IL-15 and TLR ligand adjuvants. We have found and published that nanoparticles containing S1 spike protein delivered intranasally can boost macaques primed IM with S1 in alum and result in better protection against respiratory challenge with SARS-CoV-2 than can the IM vaccine alone even though the S1-binding and neutralizing antibody levels are lower. Other mechanisms must play a role and we have found correlations with mucosal IgA, dimeric IgA, and type I interferon production in the lung, and certain types of myeloid cells. We then tested a mucosal nanoparticle boost with the B1.351 (South African) variant S1 protein to protect against this SARS-CoV-2 variant in macaques. The beta variant was the most difficult to neutralize before the appearance of the omicron variant, which had not been identified at the time we started this second study. The beta variant mucosal nanoparticle vaccine, given 1 full year after the animals had last been boosted systemically or mucosally with the original Wuhan strain, induced a 3-log increase in both IgG binding antibody in both the serum and bronchoalveolar lavage fluid (BAL, representing response in the lung). Further, it boosted the titer to the original Wuhan strain as much as to the beta variant. Neutralizing antibody titers were also similar against both virus variants. IgA and dimeric IgA to both strains were also increased. This suggests a role for original antigenic sin in determining the fine specificity of antibodies at the time of first primary vaccination. When challenged, the animals were well protected against intranasal challenge with the beta variant SARS-CoV-2. Thus, a variant intranasal vaccine can induce strong protective immunity in the lungs and nasal cavity and eliminate virus from these sites. These studies suggest that a human intranasal nanoparticle COVID-19 vaccine given to people who had been previously immunized systemically with one of the approved vaccines, could improve protection against infection and reduce the risk of forward transmission to others by reducing intranasal virus, which is especially a problem with the delta and omicron variants This second NHP study was also published. In addition, three more studies have been carried out in a hamster model, as hamsters get COVID disease more like humans. The intranasal vaccine was able to markedly reduce weight loss in the immunized animals compared to controls, implying the prevention of disease, and reduce virus particles in the lungs. It could also more effectively reduce virus in the oropharynx of hamsters than the S1 protein in alum, an important accomplishment that would be expected to reduce forward transmission. To test this hypothesis, we have now done a forward transmission study in hamsters. In this study, we also examined priming with one of the licensed mRNA vaccines to compare. Groups of hamsters were vaccinated first IM with the Moderna mRNA spike protein vaccine. Then half of them were boosted with the same mRNA vaccine IM, but the other half were boosted with the nasal nanoparticle S1 vaccine incorporating the CpG/PolyI:C & IL15 adjuvants. All the animals were challenged with SARS-CoV-2, including a group that was not immunized at all. Then, they were cohoused with naive hamsters separated only by a screen that allowed air flow between them but did not allow touching. The naive hamsters were then monitored for infection. The intranasal nanoparticle vaccine boost was substantially superior to a second IM dose of the mRNA vaccine in preventing forward transmission to the co-housed naive hamsters! This finding is critical as it demonstrates that an intranasal boost with an effective vaccine such as the one we have developed is needed to prevent COVID-19 infections from spreading, the key public health goal. Gender effects: In the hamsters, we observed that female hamsters were better protected against SARS-CoV-2 infection and disease than males, but reagents are not available to study the hamster immune response in detail. To determine the mechanism, we carried out similar studies in mice and confirmed the gender difference in mice as well. Studies are in progress to identify the mechanistic differences in immune response between the male and female mice, which could be important in optimizing efficacy of human vaccines. In vivo studies in mice: In wild type B6 mice, we have immunized with recombinant spike protein S1, S1+S2, or RBD in several different adjuvants to determine the best formulation. The best combination so far is S1 antigen with IL-15 + ligands for TLR3 and 9, for both antibody and T cell responses, but the runner up was S1 with GM-CSF and IL-12. Studies are in progress to determine which components contribute the most to protection and whether they induce qualitatively different types of immune responses. The DNA vaccine with spike protein coupled to a chemokine has been constructed and initial results show that it can induce a strong CD8 T cell response. Human cell lines: We have received the immortalized human lung epithelial cell lines, which express ACE2, from John Minna at UTSW, as well as some of his non-small-cell lung cancer cell lines that also express ACE2. We have obtained an antibody to ACE2 to verify expression. Initial results show that omega-3 fatty acids and cholesterol differentially affect ACE2 expression on lung cancer cells as well as TMPRSS2 expression and may help explain how diet and obesity as well as lung cancer can affect susceptibility to SARS-CoV-2.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><2019-nCoV vaccine><2019-nCoV variant><2019-nCoV variant forms><2019-nCoV variant strains><7S Gamma Globulin><ACE2><Adjuvant><Affect><Air Movements><Airway challenge><Airway mucosa><Alum Adjuvant><Animal Model><Animal Models and Related Studies><Animals><Antibodies><Antibody Specificity><Antibody titer measurement><Antigens><Appearance><B-Cell Epitopes><B-Lymphocyte Epitopes><B.1.1.529><B.1.617.2><Binding><Bio-Informatics><Bioinformatics><Blood Serum><Body Weight decreased><Bronchoalveolar Lavage Fluid><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><COVID><COVID-19><COVID-19 S protein><COVID-19 infection><COVID-19 predisposition><COVID-19 prevention><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 susceptibility><COVID-19 vaccine><COVID-19 variant><COVID-19 variant forms><COVID-19 variant strains><COVID-19 virus><COVID-19 virus infection><COVID-19 vulnerability><COVID19><COVID19 S protein><COVID19 infection><COVID19 prevention><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 vaccine><COVID19 virus><CV-19><CV19><Cancer cell line><Cell Body><Cell Line><CellLine><Cells><Chemotactic Cytokines><Cholesterol><CoV disease><CoV-2><CoV2><Collaborations><Coronavirus disease 2019 predisposition><Coronavirus disease 2019 susceptibility><Coronavirus disease 2019 vulnerability><Coupled><Cricetinae><DNA Vaccines><Data><Delta variant><Dendritic Cells><Diet><Disease><Disorder><Dose><Edodekin Alfa><Electroporation><Engineering><Epithelial Cells><Epitheliasin Gene><FDA EUA><FDA Emergency Use Authorization><Female><Food and Drug Administration EUA><Food and Drug Administration Emergency Use Authorization><Formulation><GM-CSF><Gender><Granulocyte-Macrophage Colony-Stimulating Factor><HIV vaccine><HIV/AIDS Vaccines><Hamsters><Hamsters Mammals><Histamine-Producing Cell-Stimulating Factor><Homologous Chemotactic Cytokines><Human><Human Cell Line><IL-12><IL-15><IL12><IL15><IL15 Protein><IgA><IgG><Immune><Immune response><Immunes><Immunity><Immunize><Immunoglobulin A><Immunoglobulin G><Immunological response><In Vitro><Industrialization><Infection><Infection prevention><Innate Immunity><Intercrines><Interferon Type I><Interleukin-12><Interleukin-15><Interleukin-15 Precursor><Licensing><Ligands><Lung><Lung Respiratory System><M mulatta><M. mulatta><MGC9721><Macaca><Macaca mulatta><Macaque><Malignant Tumor of the Lung><Malignant neoplasm of lung><Medical center><Messenger RNA><Methods><Mice><Mice Mammals><Modeling><Modern Man><Molecular Interaction><Molgramostin><Monitor><Morbidity><Morbidity - disease rate><Mucosa><Mucosal Immunity><Mucosal Tissue><Mucous Membrane><Murine><Mus><Myeloid Cells><NKSF><NSCLC><NSCLC - Non-Small Cell Lung Cancer><Naked DNA Vaccines><Nasal><Nasal Passages Nose><Nasal cavity><Native Immunity><Natural Immunity><Natural Killer Cell Stimulatory Factor><Non-Small Cell Lung Cancer><Non-Small-Cell Lung Carcinoma><Non-Specific Immunity><Nonsmall Cell Lung Carcinoma><Nonspecific Immunity><Nose><Obesity><Omega-3 Fatty Acids><Omega-3 PUFA><Omega-3 Polyunsaturated Fatty Acid><Omega3><Omicron variant><Oropharyngeal><Oropharynx><Oropharynxs><PRSS10><Persons><Predisposed to COVID-19><Predisposed to SARS-CoV-2><Predisposed to Severe acute respiratory syndrome coronavirus 2><Prevent infection><Production><Proteins><Public Health><Publishing><Pulmonary Cancer><Pulmonary malignant Neoplasm><RNA vaccine><RNA-based vaccine><Reagent><Receptor Protein><Recombinant DNA Vaccines><Recombinants><Respiratory Epithelium><Respiratory Mucosa><Respiratory System, Nose, Nasal Passages><Rhesus Macaque><Rhesus Monkey><Risk Reduction><Role><Route><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 B.1.1.529><SARS-CoV-2 B.1.617.2><SARS-CoV-2 S protein><SARS-CoV-2 delta><SARS-CoV-2 infection><SARS-CoV-2 omicron><SARS-CoV-2 omicron variant><SARS-CoV-2 predisposition><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV-2 susceptibility><SARS-CoV-2 vaccine><SARS-CoV-2 variant><SARS-CoV-2 variant forms><SARS-CoV-2 variant strains><SARS-CoV-2 vulnerability><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 infection><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><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><SIS cytokines><SIV Vaccines><Serum><Severe Acute Respiratory Coronavirus 2><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 coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 predisposition><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome coronavirus 2 susceptibility><Severe acute respiratory syndrome coronavirus 2 vaccine><Severe acute respiratory syndrome coronavirus 2 vulnerability><Severe acute respiratory syndrome related corona virus 2><Site><South African><Strains Cell Lines><Structure of respiratory epithelium><T cell response><T-Cells><T-Lymphocyte><T8 Cells><T8 Lymphocytes><TC-GM-CSF><TLR3><TLR3 gene><TMPRSS2><TMPRSS2 gene><Testing><Texas><Time><Toll-Like Receptor 3><Touch><Touch sensation><Transgenic Mice><Transmission><Tumor-Cell Human GM Colony-Stimulating Factor><Universities><Vaccinated><Vaccination><Vaccines><Variant><Variation><Vascular Endothelial Cell><Veiled Cells><Viral><Viral Vaccines><Virion><Virus><Virus Particle><Weight Loss><Weight Reduction><Work><Wuhan coronavirus><adiposity><air flow><airflow><airway epithelium><alum><aluminum sulfate><angiotensin converting enzyme 2><angiotensin converting enzyme II><antibody titering><body weight loss><chemoattractant cytokine><chemokine><compare to control><comparison control><corona virus disease><corona virus disease 2019><corona virus disease 2019 vaccine><coronavirus disease><coronavirus disease 2019><coronavirus disease 2019 S protein><coronavirus disease 2019 infection><coronavirus disease 2019 prevention><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 vaccine><coronavirus disease 2019 variant><coronavirus disease 2019 variant forms><coronavirus disease 2019 variant strains><coronavirus disease 2019 virus><coronavirus disease-19><coronavirus disease-19 prevention><coronavirus disease-19 vaccine><coronavirus disease-19 virus><coronavirus infectious disease-19><corpulence><cultured cell line><diets><dimer><disease control><disease prevention><disorder control><disorder prevention><electroporative delivery><emergency use authorization><experience><gender difference><gender-associated difference><gene electrotransfer><granulocyte macrophage colony stimulating factor><hCoV19><health goals><host response><human immunodeficiency virus vaccine><immune system response><immunogen><immunogenicity><immunopathology><immunoresponse><improved><in vivo><indicated prevention><indicated preventive interventions><indicated preventive measure><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><lung cancer><lung cancer cell><mRNA><mRNA vaccine><mRNA-based vaccine><male><model of animal><mucosal vaccine><n-3 Fatty Acids><nCoV vaccine><nCoV-19 vaccine><nCoV19 vaccine><nCoV2><nano particle><nano-sized particle><nanoparticle><nanosized particle><neutralizing antibody><non-human primate><nonhuman primate><omega-3><omicron variant of COVID-19><omicron variant of SARS-CoV-2><oral pharyngeal><pre-clinical study><preclinical study><predisposed to Coronavirus disease 2019><prevent><prevent COVID-19><prevent COVID19><prevent coronavirus disease 2019><preventing><prevention directed at individuals><pulmonary><receptor><receptor binding><receptor bound><reduce risk><reduce risks><reduce that risk><reduce the risk><reduce these risks><reduces risk><reduces the risk><reducing risk><reducing the risk><respiratory challenge><respiratory tract epithelium><response><risk-reducing><severe acute respiratory syndrome coronavirus 2 B.1.1.529><severe acute respiratory syndrome coronavirus 2 B.1.617.2><severe acute respiratory syndrome coronavirus 2 variant><severe acute respiratory syndrome coronavirus 2 variant forms><severe acute respiratory syndrome coronavirus 2 variant strains><simian immunodeficiency virus vaccines><social role><susceptible to COVID-19><susceptible to Coronavirus disease 2019><susceptible to SARS-CoV-2><susceptible to Severe acute respiratory syndrome coronavirus 2><tactile sensation><thymus derived lymphocyte><transmission process><vaccination study><vaccination trial><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 candidate><vaccine for novel coronavirus><vaccine study><vaccine trial><variants of concern><viral transmission><virus transmission><wt-loss><ω-3 fatty acids>