Quantifying human natural killer cell-mediated direct killing of target cells
Presenter Type
UNO Undergraduate Student
Major/Field of Study
Biology
Other
Biology
Advisor Information
Department of Biology, Assistant Professor, PhD
Location
MBSC302 - U
Presentation Type
Oral Presentation
Start Date
24-3-2023 2:30 PM
End Date
24-3-2023 3:45 PM
Abstract
Peripheral blood mononuclear cells (PBMCs) include cells that are essential to the functions of the immune system, both in the innate and adaptive immune responses. Natural killer (NK) cells are a primary cell type responsible for executing innate responses. NK cells can recognize and kill infected or malignant cells without the need for memory functions or antigen specificity. However, NK cells can also destroy their targets using antibody-dependent cellular cytotoxicity, or ADCC (the topic of a separate presentation by fellow lab member, Cami Bisson). Direct killing requires only that target cells be recognized as “diseased” by the natural killer cells. Typically, this recognition is made based on target cells lacking expression of a normally present cell surface marker (i.e., MHC-I). Following the target cell recognition, NK cells initiate a cytotoxic response that lyses the target cell. Our lab has developed a testing system, referred to as the Natural Killer cell Simultaneous ADCC and Direct Killing Assay (NK-SADKA), that allows for the analysis of both ADCC and direct killing. The same donor samples are used for both the direct killing and ADCC portions of the assay to control for donor response variation. Within the direct killing portion of the assay (the focus of this presentation), MHC-I-deficient human leukemia cells (e.g., K562s) are labeled with a fluorescent dye to distinguish them from the unlabeled human NK effector cells. Target and effector cells are combined (5:1 effector to target ratio) and co-incubated (2 hours). Killing capacity is analyzed using a flow cytometer. Our direct killing arm of the NK-SADKA is functioning properly, as demonstrated by our data showing substantial killing following the coincubation relative to the viability of targets or effectors incubated alone. Data-to-date will be presented. Our NK-SADKA, including the direct killing arm described herein, will have impacts on clinical trial interpretations and designs, particularly when those trials are using interventions (e.g., immunotherapies) intended to alter NK cell killing capacities.
Scheduling
1-2:15 p.m., 2:30 -3:45 p.m.
Quantifying human natural killer cell-mediated direct killing of target cells
MBSC302 - U
Peripheral blood mononuclear cells (PBMCs) include cells that are essential to the functions of the immune system, both in the innate and adaptive immune responses. Natural killer (NK) cells are a primary cell type responsible for executing innate responses. NK cells can recognize and kill infected or malignant cells without the need for memory functions or antigen specificity. However, NK cells can also destroy their targets using antibody-dependent cellular cytotoxicity, or ADCC (the topic of a separate presentation by fellow lab member, Cami Bisson). Direct killing requires only that target cells be recognized as “diseased” by the natural killer cells. Typically, this recognition is made based on target cells lacking expression of a normally present cell surface marker (i.e., MHC-I). Following the target cell recognition, NK cells initiate a cytotoxic response that lyses the target cell. Our lab has developed a testing system, referred to as the Natural Killer cell Simultaneous ADCC and Direct Killing Assay (NK-SADKA), that allows for the analysis of both ADCC and direct killing. The same donor samples are used for both the direct killing and ADCC portions of the assay to control for donor response variation. Within the direct killing portion of the assay (the focus of this presentation), MHC-I-deficient human leukemia cells (e.g., K562s) are labeled with a fluorescent dye to distinguish them from the unlabeled human NK effector cells. Target and effector cells are combined (5:1 effector to target ratio) and co-incubated (2 hours). Killing capacity is analyzed using a flow cytometer. Our direct killing arm of the NK-SADKA is functioning properly, as demonstrated by our data showing substantial killing following the coincubation relative to the viability of targets or effectors incubated alone. Data-to-date will be presented. Our NK-SADKA, including the direct killing arm described herein, will have impacts on clinical trial interpretations and designs, particularly when those trials are using interventions (e.g., immunotherapies) intended to alter NK cell killing capacities.
Additional Information (Optional)
Could you place me after a presentation titled, "Quantifying human natural killer cell-mediated antibody-dependent cellular cytotoxicity" by Cami Bisson? Also I have yet to finish, but would I send in the powerpoint that I would hope to use?