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Principal Investigator: Anna Karolina Palucka
Organization: JACKSON LABORATORY
Fiscal Year: 2023
Award: $291,555
Funding agency: NIH Office of the Director
PROJECT SUMMARY
Although humanized mice have been successfully used for in vivo studies of HIV infection, cancer and
immunotherapies, human naïve and adaptive immune responses remain suboptimal, with limited cross-reactivity
between murine and human cytokines considered as a key contributing factor. As a result, the human mucosal
immune system is not fully developed, and several components are missing, including the presence of human
lung epithelial cells in their natural lung environment. To address these research gaps, we propose a combined
genetic and cellular editing approach to develop the next generation of humanized mice for studies of human
mucosal immunity. Our approach is structured in two aims: in Aim 1, we will construct and credential hNSGF-
SGM3-IL6-TSLP-TSLPR mice for human immune cell development. We will generate mice expressing a
complete human TSLP receptor and examine engraftment with human hematopoietic progenitor cells (HPCs)
by measuring cellular composition of immune cells in the bone marrow, spleen, gut, airways, lungs, and blood.
To examine human immune function, we will use a mucosal formulation of influenza vaccine (Flumist) as an
immune trigger and antigen source as we have done in the past and analyze cytokine responses in the blood
and spleen; human cell migration to mucosal sites, spleen, and bone marrow; and induction of vaccine antigen-
specific T and B cells in the spleen. In Aim 2, we will construct and credential humanized airway epithelium in
hNSGF-SGM3-IL6-TSLP-TSLPR mice. We will examine the development of human airway epithelium upon
transplant of human bronchial epithelial progenitor cells in NSGF-SGM3-IL6-TSLP-TSLPR mice (humanized or
not with HPCs from the same donor) by tissue immunofluorescence staining of human-specific targets including
HLA class I. To establish the functionality of human airway epithelial cells, we will challenge mice with live
influenza virus and measure the induction of species-specific alarmins including IL-33; type I interferon signature
in human epithelial cells as a measure of their functionality in vivo, as well as tissue composition and attraction
of human immune cells to human airway epithelium. Our hypothesis is that a complete TSLP signaling pathway
and human epithelial cells will improve human mucosal immunity in humanized mice by facilitating the crosstalk
between stromal cells and immune cells. This model can then be used for i. mucosal vaccine development, and
ii. studies of other respiratory viruses including SARS-CoV-2, which requires human-specific receptors to
establish infection. Thus, once credentialed, our model will lay the groundwork for future mechanistic studies of
mucosal immunity in vivo in the context of genetically variable donors as well as enable studies of mucosal
adjuvants, allergens, vaccines, and biologicals.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><Address><Adjuvant><Airway infections><Allergens><Allergic asthma><Allergy><Antibodies><Antigens><Autoregulation><B blood cells><B cell><B cell differentiation factor><B cell stimulating factor 2><B cells><B-Cell Activation><B-Cell Differentiation Factor><B-Cell Differentiation Factor Gene><B-Cell Differentiation Factor-2><B-Cell Stimulatory Factor 2 Gene><B-Cell Stimulatory Factor-2><B-Cells><B-Lymphocytes><B-cell><BCDF><BSF-2><BSF-2 Gene><BSF2><BSF2 Gene><Beta-2 Gene Interferon><Biological><Blood><Blood Plasma Cell><Blood Precursor Cell><Blood Reticuloendothelial System><Blood monocyte><Body Tissues><Bone Marrow><Bone Marrow Reticuloendothelial System><Bronchi><CAIV-T><CD127><CD34><CD34 gene><CDW127><COVID-19 virus><COVID19 virus><CRISPR><CRISPR/Cas system><CSF-1><Cancers><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Function><Cell Interaction><Cell Locomotion><Cell Migration><Cell Movement><Cell Process><Cell Signaling><Cell physiology><Cell-to-Cell Interaction><Cells><Cellular Function><Cellular Immune Function><Cellular Migration><Cellular Motility><Cellular Physiology><Cellular Process><Clinic><Clustered Regularly Interspaced Short Palindromic Repeats><CoV-2><CoV2><Colon><Colony-Stimulating Factor 1><Credentialing><Dendritic Cells><Development><Disease><Disorder><ELISPOT><Engraftment><Environment><Epithelial Cells><Epithelium><Extrinsic asthma><FLK2><FLT3><FLT3 gene><FMS-like tyrosine kinase 3><Fibroblasts><FluMist><Fms-Related Tyrosine Kinase 3><Formulation><Future><Generations><Genetic><Genetic Engineering><Genetic Engineering Biotechnology><Genetic Engineering Molecular Biology><HIV Infections><HPCA1><HPGF><HSF Gene><HTLV-III Infections><HTLV-III-LAV Infections><Helper Cells><Helper T-Cells><Helper T-Lymphocytes><Helper-Inducer T-Cells><Helper-Inducer T-Lymphocyte><Hematopoietic><Hematopoietic Progenitor Cells><Hematopoietic stem cells><Hepatocyte Stimulatory Factor Gene><Hepatocyte-Stimulating Factor><Homeostasis><Human><Human T-Lymphotropic Virus Type III Infections><Hybridoma Growth Factor><Hybridoma Growth Factor Gene><Hypersensitivity><IFN-beta 2><IFNB2><IFNB2 Gene><IL-6><IL-6 Gene><IL-7R alpha chain><IL-7R-alpha><IL-7Ralpha><IL-7Rα><IL6><IL6 Protein><IL6 gene><IL7R><IL7R gene><Immune><Immune mediated therapy><Immune response><Immune system><Immunes><Immunodeficient Mouse><Immunofluorescence><Immunofluorescence Immunologic><Immunological response><Immunologically Directed Therapy><Immunotherapy><In Vitro><Inducer Cells><Inducer T-Lymphocytes><Infection><Inflammation><Inflammatory><Influenza Vaccines><Influenza Virus><Innate Immune Response><Interferon Type I><Interleukin 6 (Interferon, Beta 2) Gene><Interleukin-6><Interleukin-6 Gene><Interleukins><Intracellular Communication and Signaling><KI mice><KO mice><Knock-in><Knock-in Mouse><Knock-out Mice><Knockout Mice><Leiomyocyte><Liver><Lung><Lung Respiratory System><Lung damage><Lymphatic Tissue><Lymphoid><Lymphoid Tissue><M-CSF><MGF Stem Cell Factor><MGI-2><Macrophage><Macrophage Colony-Stimulating Factor><Malignant Neoplasms><Malignant Tumor><Marrow monocyte><Mast Cell Growth Factor><Measures><Mice><Mice Mammals><Modeling><Modern Man><Motility><Mucosa><Mucosal Immune System><Mucosal Immunity><Mucosal Tissue><Mucous Membrane><Murine><Mus><Myeloid Cells><Myeloid Differentiation-Inducing Protein><Mφ><NIAID><NIH><National Institute of Allergy and Infectious Disease><National Institutes of Health><Natural regeneration><Null Mouse><PBMC><Peripheral Blood Mononuclear Cell><Phenotype><Physiological Homeostasis><Plasma Cells><Plasmacytes><Plasmacytoma Growth Factor><Play><Population><Pre-Clinical Model><Preclinical Models><Property><Proteins><Receptor Protein><Recombinant DNA Technology><Regeneration><Research><Respiratory Epithelium><Respiratory Infections><Respiratory Tract Infections><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV2><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><STK-1 kinase><STK1><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-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 related corona virus 2><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Smooth Muscle Cells><Smooth Muscle Myocytes><Smooth Muscle Tissue Cell><Source><Spleen><Spleen Reticuloendothelial System><Staining method><Stains><Steel Factor><Stem Cell Factor><Stem Cell Tyrosine Kinase 1><Stromal Cells><Structure><Structure of respiratory epithelium><Subcellular Process><System><Systemic infection><T cell response><T-Cells><T-Lymphocyte><TSLP><TSLP gene><Testing><Thymic Stromal Lymphopoietin><Thymic Tissue><Tissues><Transgenic Organisms><Transplantation><United States National Institutes of Health><Vaccine Antigen><Vaccines><Veiled Cells><Virus><Wuhan coronavirus><activated B cells><adaptive immune response><adaptive immunity><airway colonization><airway epithelium><airway smooth muscle><alpha chain interleukin-7 receptor><antigen-specific T cells><atopic asthma><biologic><biological signal transduction><blood stem cell><bronchial epithelium><c-kit Ligand><cell motility><cell regeneration><cell type><cellular regeneration><coronavirus disease 2019 virus><coronavirus disease-19 virus><cross reactivity><cytokine><develop a vaccine><develop vaccines><development of a vaccine><developmental><enzyme linked immunospot assay><epithelial progenitor cell><epithelial stem cell><extrinsic allergic asthma><fetal><fetal liver kinase-2><fetal liver kinase-3><flu infection><flu vaccine><flu virus infection><flu virus vaccine><genetic approach><genetic strategy><genetically engineered><hCoV19><hematopoietic progenitor><hematopoietic stem progenitor cell><hemopoietic><hemopoietic progenitor><hemopoietic stem cell><hepatic body system><hepatic organ system><host response><human stem cells><humanized mice><humanized mouse><immune function><immune system response><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immuno therapy><immunogen><immunoresponse><improved><in vivo><infected with flu><infected with flu virus><infected with influenza><infected with influenza virus><influenza infection><influenza virus infection><influenza virus vaccine><influenzavirus><injury to tissue><interferon beta 2><kit Ligand><knockin><knockin mice><lung injury><malignancy><monocyte><mouse development><mouse model><mucosal site><mucosal vaccine><murine model><nCoV2><neoplasm/cancer><next generation><novel><overexpress><overexpression><permissiveness><plasmocyte><pulmonary><pulmonary damage><pulmonary injury><pulmonary tissue damage><pulmonary tissue injury><receptor><regenerate><respiratory colonization><respiratory smooth muscle><respiratory tract epithelium><respiratory virus><response><thymus derived lymphocyte><tissue injury><transgenic><translational immunology><transplant><vaccine against flu><vaccine against influenza><vaccine development>