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Principal Investigator: Kerry S Campbell
Organization: THOMAS JEFFERSON UNIVERSITY
Fiscal Year: 2024
Award: $498,670
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary
NK cells are capable of killing virus-infected and cancer cells and, therefore, play an important role in innate
host defense. Unlike cytotoxic T lymphocytes (CTL), whose cytolytic activity is induced by productive TCR
engagement with cognate pMHC ligands, activation of NK cells is regulated by a set of activating and inhibitory
receptors. The net result of the signals induced by these receptors determines the extent of NK cell effector
activities such as INF-γ production and granule-mediated cytotoxicity. One of the main mechanisms of NK cell-
mediated cytolytic activity depends on antibodies and is termed antibody-dependent cell-mediated cytotoxicity
(ADCC). ADCC is induced by FcγRIIIa (CD16) receptors that do not directly recognize antigens on the target
cells, but interact with Fc fragments of IgG antibodies specifically bound to the antigens on the surface of target
cells. Available evidence, including ours, strongly suggests that ligand-mediated clustering of activating
receptors on cytotoxic lymphocytes regulates kinetics of intracellular signaling, formation of a highly ordered
synaptic interface, kinetics of granule delivery, and efficiency of target cell lysis (1-6). The receptor clustering
can be further modulated by antigen co-clustering with other membrane ligands on target cells, which interact
with inhibitory receptors or adhesion molecules on the surface of NK cells. We hypothesize that differences
in FcγRIIIa (CD16) clustering/co-clustering could have significant impact on receptor-mediated
signaling influencing efficiency of ADCC. Thus, the goal of this project is to study how homotypic and
heterotypic ligand clustering regulates ability of the ligands to cooperate in the engagement of CD16, inhibitory
receptors and adhesion molecules to modulate efficiency of ADCC. We will use well-characterized CD16.NK92
cells that we have developed to model functions of NK cells (7-12) and freshly activated human NK cells to
investigate the role of CD16 clustering/co-clustering with other receptors. To achieve this goal, we will exploit
fluorescent nanoparticles (NiNLPs) that can be loaded with ligands for CD16, adhesion molecules, and
inhibitory receptors at various ratios and densities, and will examine the binding kinetics of these model
membrane clusters to NK cells as well as the kinetics of induced intracellular Ca2+ signaling. We will also
utilize planar lipid bilayers presenting the same ligands, either dispersed in the bilayers or co-localized by
cross-linking with the iDimerize system, and will analyze how cross-linking of the ligands will influence structure
of the synaptic interface and the kinetics of NK cell degranulation. The expected results will provide
significantly improved understanding of the mechanisms controlling cytolytic activity of primary and activated
human NK cells to design new NK cell-based immunotherapeutic strategies to improve efficiency of ADCC to
treat viral infections, autoimmune diseases, and cancer.
Terms: <7S Gamma Globulin><Ab-dependent cellular cytotoxicity><Adhesion Molecule><Affect><Affinity><Antibodies><Antigen Binding Fragment><Antigens><Autoimmune Diseases><Avidity><Binding><Bp35><C2B8 Monoclonal Antibody><CD16><CD16B><CD20><CD54 Antigens><CD8><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><CD8B><CD8B1><CD8B1 gene><Cancers><Cell Adhesion Molecule Gene><Cell Adhesion Molecules><Cell Body><Cell Communication and Signaling><Cell Degranulation><Cell Maturation><Cell Signaling><Cell surface><Cell-Mediated Lympholytic Cells><Cells><Cellular Immune Function><Closure by Ligation><Communicable Diseases><Cytolysis><Cytolytic T-Cell><Cytoplasmic Granules><Cytotoxic T Cell><Cytotoxic T-Lymphocytes><Cytotoxic cell><Data><Development><Diffusion><Disease><Disorder><Dissociation><Effector Cell><Environment><FCGR3B><FCGR3B gene><Fab Fragments><Fab Immunoglobulins><Fc Fragments><Fc Immunoglobulins><Fc Receptor III-1><Fc gamma IIIb receptor><Fc-Gamma RIII-Beta><Fc-Gamma RIIIB><FcRIIIB><Goals><Host Defense><Human><ICAM-1><IgG><IgG Fc Receptor IIIB><IgG1><IgG3><Immune mediated therapy><Immunoglobulin G><Immunoglobulin, F(ab) Fragment><Immunologic Receptors><Immunological Receptors><Immunologically Directed Therapy><Immunotherapeutic agent><Immunotherapy><Infection><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Integrins><Integrins Extracellular Matrix><Intercellular adhesion molecule 1><Intracellular Communication and Signaling><K lymphocyte><Kinetics><LYT3><Leu-16><Ligands><Ligation><Lipid Bilayers><Low Affinity IgG Fc Receptor IIIB><Low Affinity Immunoglobulin Gamma Fc Region Receptor III-B><Lymphatic cell><Lymphocyte><Lymphocytic><Lysis><Lytotoxicity><MHC Interaction><MS4A1><MS4A1 gene><MS4A2><MabThera><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Mediating><Meditation><Membrane><Modeling><Modern Man><Molecular><Molecular Interaction><NK Cell Activation><NK Cells><NK cell immunotherapy><NK cell-based immunotherapy><NK cytotoxicity><Names><Natural Killer Cell Activation><Natural Killer Cell Immunotherapy><Natural Killer Cell toxicity><Natural Killer Cells><Natural Killer cytotoxicity><Nature><Peptide-MHC><Peptide-Major Histocompatibility Protein Complex><Peptide/MHC Complex><Play><Position><Positioning Attribute><Production><Productivity><Proteins><Receptor Cell><Receptor Protein><Rituxan><Role><Signal Induction><Signal Transduction><Signal Transduction Systems><Signaling><Structure><Surface><Synapses><Synaptic><System><T cell response><T-Cells><T-Lymphocyte><T8 Cells><T8 Lymphocytes><Testing><Therapeutic><Variant><Variation><Viral Diseases><Virus><Virus Diseases><antibody dependent cell mediated cytotoxicity><antibody dependent cytotoxicity><antibody mediated cellular cytotoxicity><antibody-dependent cell cytotoxicity><antibody-dependent cellular cytotoxicity><antibody-mediated cytotoxicity><autoimmune condition><autoimmune disorder><autoimmunity disease><biological signal transduction><cancer cell><cell adhesion protein><cell killing><crosslink><cytokine><cytotoxic><cytotoxicity><density><design><designing><developmental><diffused><diffuses><diffusing><diffusions><granule><immune drugs><immune function><immune receptor><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapeutics><immune-based therapies><immune-based treatments><immuno therapy><immunogen><immunologic therapeutics><immunological synapse formation><immunotherapeutics><immunotherapy agent><improved><killer T cell><kinetic model><lipid bilayer membrane><lymph cell><malignancy><membrane model><membrane structure><name><named><naming><nano meter scale><nano meter sized><nano particle><nano-sized particle><nanocluster><nanolipoprotein particles><nanometer scale><nanometer sized><nanoparticle><nanoscale><nanosized particle><natural killer cell based immunotherapy><neoplasm/cancer><new approaches><novel approaches><novel strategies><novel strategy><pMHC><pathogen><receptor><receptor-mediated signaling><response><rituximab><social role><synapse><synapse formation><synapse function><synaptic function><synaptogenesis><thymus derived lymphocyte><tool><tumor><viral infection><virus infection><virus-induced disease>