NK Cell Phenotyping Using Two Complementary Flow Cytometric Analysis Strategies
Presenter Type
UNO Undergraduate Student
Major/Field of Study
Biology
Author ORCID Identifier
0009-0009-0483-5228
Advisor Information
Dr. Paul W. Denton
Location
CEC RM #231
Presentation Type
Oral Presentation
Start Date
22-3-2024 9:00 AM
End Date
22-3-2024 10:15 AM
Abstract
Natural killer (NK) cells are a component of the innate immune system, which functions to kill disease cells. NK cells achieve this in one of two ways: direct killing or antibody dependent cell-mediated cytotoxicity. With stimulation/activation, NK cells can experience changes in their surface phenotype due to changes in the composition/ratio of specific proteins (markers) present on the cell surface. These changes are related to the function of a cell. For example, cytokine-producing cells and regulatory cells normally have distinct surface phenotypes. Typically, we measure phenotype changes in terms of differences in the presence/absence and frequency of a specific marker(s). We have the capacity to measure the surface phenotype changes by using fluorescently labeled antibodies that specifically bind to a given marker. Then we use a flow cytometer to detect the fluorescence from the bound antibodies. Using our laboratory’s immunophenotyping panel, we have begun examining the dynamics of these immunophenotypes, including changes that occur in the presence of cancer cells used in the laboratory as target cells for NK cell-mediated killing. Typically, we analyze our flow cytometry data in a software platform called FlowJo. My project is. To couple our FlowJo analyses with a cloud-based analysis software called Cytobank to identify additional phenotypic characteristics that can only be found using such network analysis strategies. With this analysis strategy established, our goal is to apply this approach to improve our ability to relate the functional outcomes of human NK cell-mediated killing studies with surface phenotypes. Data to date will be presented.
NK Cell Phenotyping Using Two Complementary Flow Cytometric Analysis Strategies
CEC RM #231
Natural killer (NK) cells are a component of the innate immune system, which functions to kill disease cells. NK cells achieve this in one of two ways: direct killing or antibody dependent cell-mediated cytotoxicity. With stimulation/activation, NK cells can experience changes in their surface phenotype due to changes in the composition/ratio of specific proteins (markers) present on the cell surface. These changes are related to the function of a cell. For example, cytokine-producing cells and regulatory cells normally have distinct surface phenotypes. Typically, we measure phenotype changes in terms of differences in the presence/absence and frequency of a specific marker(s). We have the capacity to measure the surface phenotype changes by using fluorescently labeled antibodies that specifically bind to a given marker. Then we use a flow cytometer to detect the fluorescence from the bound antibodies. Using our laboratory’s immunophenotyping panel, we have begun examining the dynamics of these immunophenotypes, including changes that occur in the presence of cancer cells used in the laboratory as target cells for NK cell-mediated killing. Typically, we analyze our flow cytometry data in a software platform called FlowJo. My project is. To couple our FlowJo analyses with a cloud-based analysis software called Cytobank to identify additional phenotypic characteristics that can only be found using such network analysis strategies. With this analysis strategy established, our goal is to apply this approach to improve our ability to relate the functional outcomes of human NK cell-mediated killing studies with surface phenotypes. Data to date will be presented.