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Principal Investigator: Mark Morris Davis
Organization: STANFORD UNIVERSITY
Fiscal Year: 2024
Award: $553,826
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY
In this technology development project, we want to continue the significant progress that we made in this last
granting period and take advantage of the many opportunities that we see to make human immune organoids
an indispensable system for understanding human immune responses to vaccination and infection. Fortunately,
we have made several major advances in our organoid work recently, especially our focus on spleen organoids
that we obtain through our collaboration with Donor Network West, which now regularly supplies us with both
spleens, skin, blood and lung fragments from a given donor. We have found that spleens are much more diverse
in cell types than tonsils and are easier to get responses from both RNA vaccines and novel ones(to most
individuals) such as Yellow Fever Vaccine. We will use them in Aim 1. to optimize the protocols for flu and SARS-
CoV-2 RNA vaccines, and specifically to determine how well organoid responses compare with available studies
in live humans, using our available reagents for T and B cell responses. An even deeper exploration of the BCR
and TCR repertoire will be achieved by sequence analysis, which the Boyd and Davis groups are very
experienced at. In Aim 2 we will focus on getting complete T and B cell responses with the Yellow Fever vaccine,
currently producing only IgM antibodies. In Aim 3, we will take advantage of promising results we have obtained
showing that a highly multimerized peptide-MHC reagent (60-mer) can stain specific T cells in fixed sections, as
can a reagent for specific B cells. These would be a very valuable reagents to interrogate specific lymphocytes
in the most standard type of clinical specimen. In Aim 4, we are teaming up with Drs. Kuo and Blish to use lung
and spleen organoids from the same donors to see any form of cooperativity when we infect the lungs with
influenza or SARS-CoV-2. Since we can see infiltration and rejection with co-incubated skin and tonsils/spleen
from unmatched donors, we are confident that there will cooperativity, as we expect antigen loaded dendritic
cells will migrate from infected lungs and trigger a response in nearby spleen organoids. Lastly, in Aim 5 we
suspect that there are distinct differences in the immune responses of inbred mice and humans, and so have
recently started make mouse spleen organoids so that we can analyze their vaccine responses alongside human
spleens. With the advanced single cell analysis tools available it should be relatively straightforward to identify
differences. We can then follow up those differences with gene editing using CRISPR-Cas9, as we have done
recently. We will also serve as a resource for all three of the main projects, using gene editing and other methods
with organoids for Dr. Wang and Khatri’s projects, as well has interrogating spleen organoids from high and low
BMI individuals in collaboration with those projects and for quickly testing novel immunogens with Dr. Barnes.
Terms: <19S Gamma Globulin><2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV vaccine><2019-nCoV variant><2019-nCoV variant forms><2019-nCoV variant strains><7S Gamma Globulin><Active Follow-up><Algorithms><Aliquot><Anti-CD40><Antibodies><Antigen Receptors><Antigenic Determinants><Antigens><Assay><Attenuated><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><BMI><BMI percentile><BMI z-score><Binding><Binding Determinants><Bioassay><Biological Assay><Biopsy><Biotinylation><Blood><Blood Reticuloendothelial System><Body Tissues><Body mass index><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><COVID-19 vaccine><COVID-19 variant><COVID-19 variant forms><COVID-19 variant strains><COVID-19 virus><COVID19 virus><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><CSIF><CSIF-10><Cas nuclease technology><Cell Body><Cells><Class Switching><Class Switchings><Clinical><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Co-culture><CoV-2><CoV2><Cocultivation><Coculture><Coculture Techniques><Collaborations><Cytokine Synthesis Inhibitory Factor><DNA><Dendritic Cells><Deoxyribonucleic Acid><Epitopes><Exposure to><Fixation><Formalin><Gene Transcription><Genes><Genetic Transcription><Grant><Grippe><Human><IL-10><IL10><IL10A><IgG><IgM><Immune><Immune response><Immunes><Immunochemical Immunologic><Immunoglobulin Class Switching><Immunoglobulin Class Switchings><Immunoglobulin G><Immunoglobulin M><Immunologic><Immunological><Immunological response><Immunologically><Immunologics><In Vitro><Inbred Mouse><Individual><Infection><Infiltration><Influenza><Influenza Vaccines><Interleukin 10 Precursor><Interleukin-10><Isotype Switching><Isotype Switchings><Left><Link><Lung><Lung Respiratory System><Lymphatic cell><Lymphocyte><Lymphocytic><Methods><Mice><Mice Mammals><Modern Man><Molecular Interaction><Murine><Mus><Norway><Organoids><Peptide-MHC><Peptide-Major Histocompatibility Protein Complex><Peptide/MHC Complex><Phenotype><Process><Protocol><Protocols documentation><Quetelet index><RNA Expression><RNA vaccine><RNA-based vaccine><Reagent><Research Resources><Research Specimen><Resources><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 vaccine><SARS-CoV-2 variant><SARS-CoV-2 variant forms><SARS-CoV-2 variant strains><SARS-CoV2><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><SEQ-AN><Sequence Analyses><Sequence Analysis><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 vaccine><Severe acute respiratory syndrome related corona virus 2><Skin><Specificity><Specimen><Spleen><Spleen Reticuloendothelial System><Staining method><Stains><Standardization><Stimulus><Strepavidin><Streptavidin><Survey Instrument><Surveys><System><T cell response><T-Cells><T-Lymphocyte><T-cell receptor repertoire><T8 Cells><T8 Lymphocytes><TCR repertoire><Technology><Testing><Time><Tissues><Tonsil><Transcription><Vaccines><Veiled Cells><Viral Diseases><Virus Diseases><Work><Wuhan coronavirus><Yellow Fever Vaccine><active followup><attenuate><attenuates><cell type><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 vaccine><coronavirus disease-19 virus><evaluate vaccines><experience><flu><flu vaccine><flu virus vaccine><follow up><follow-up><followed up><followup><hCoV19><host response><human subject><immune response to vaccination><immune response to vaccines><immune system response><immunogen><immunoresponse><improved><in vivo><influenza virus vaccine><lumazine><lymph cell><mRNA vaccine><mRNA-based vaccine><migration><monomer><nCoV vaccine><nCoV-19 vaccine><nCoV19 vaccine><nCoV2><new vaccines><next generation vaccines><novel><novel vaccines><outcome following vaccination><outcome following vaccine><pMHC><pathogen><prevent><preventing><pulmonary><response><result following vaccination><result following vaccine><sample fixation><self assembly><severe acute respiratory syndrome coronavirus 2 variant><severe acute respiratory syndrome coronavirus 2 variant forms><severe acute respiratory syndrome coronavirus 2 variant strains><single cell analysis><single cell technology><tech development><technology development><thymus derived lymphocyte><tissue fixing><tonsillar><tool><vaccination outcome><vaccination result><vaccine against 2019-nCov><vaccine against COVID-19><vaccine against SARS-CoV-2><vaccine against SARS-coronavirus-2><vaccine against Severe Acute Respiratory Syndrome CoV 2><vaccine against Severe acute respiratory syndrome coronavirus 2><vaccine against flu><vaccine against influenza><vaccine against yellow fever><vaccine associated immune response><vaccine candidates against SARS-CoV-2><vaccine efficacy><vaccine evaluation><vaccine failure><vaccine for novel coronavirus><vaccine immune response><vaccine immunogenicity><vaccine induced immune response><vaccine outcome><vaccine response><vaccine responsiveness><vaccine result><vaccine screening><vaccine testing><vaccine-induced response><vaccines preventing COVID><vaccines to prevent COVID><viral infection><virus infection><virus-induced disease><yellow fever virus vaccine>