Date: Wednesday, October 6, 2021
Time: 7 a.m. PT | 10 a.m. ET |4 p.m. CET
Cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells have become attractive therapeutic targets for cancer immunotherapy because of their efficient mechanisms for killing cancerous cells. Both effector CTLs and mature NK cells are endowed with secretory granules containing pre-formed cytolytic proteins that once released at the immunological synapse, initiate a cascade of reactions leading to the apoptosis of the target cell.
This webcast will describe a simplified intracellular staining flow cytometry workflow for identification of circulating T and NK cells with cytolytic potential. In this workflow, a 16-color antibody panel is used to measure cytolytic proteins, including granzyme K, granzyme B and perforin, enabling functional classification of various cell subsets within conventional and unconventional T cells and NK cells. Red blood cell lysis and cell fixation are carried out directly in whole blood and yet enable the detection of low frequency cytotoxic lymphocyte populations.
Learn how to:
- Efficiently perform intracellular staining of immune cells directly in whole blood for flow cytometry experiments
- Obtain optimal resolution of small subsets of circulating NK cells and T cells with cytolytic potential
- Integrate tools found in FlowJo™ v10.7.2 Software for simple visualization and exploration of the data set
Dr. Gisele Baracho
Staff Scientist, Applied Research and Technology
Gisele Baracho earned her PhD in Immunology from the University of São Paulo, Brazil. During her graduate studies, she focused on the characterization of cohorts of patients with primary immunodeficiency disorders. Because of her keen interest in immunopathological disorders and cancer, she became a postdoctoral fellow at Sanford Burnham Prebys Medical Discovery Institute. Her postdoctoral studies focused on understanding molecular mechanisms that regulate normal B cell development and tumorigenesis in mouse models. She joined BD Biosciences as a Sr. Scientist to work on a variety of cell-based and flow cytometry assays and to support the release of new reagents. Currently, she is a Staff Scientist in the Applied Research and Technology team within Medical and Scientific Affairs, responsible for the development of innovative flow cytometry applications. She also completed a 3-month internship at New York University Langone Medical Center, where she acquired further expertise in high-dimensional single-cell analysis.