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A novel flow cytometric assay for evaluating cell-mediated cytotoxicity
Mark W Burkett1, Kimberly A Shafer-Weaver, Susan Strobl
1Laboratory of Cell-Mediated Immunity, Clinical Services Program, SAIC-Frederick, Inc., Frederick, Maryland 21702, USA.
Journal of Immunotherapy (Hagerstown, Md. : 1997)
|July 8, 2005
Summary
This study introduces a novel assay to measure cell-mediated cytotoxicity by simultaneously assessing effector cell degranulation (CD107a) and target cell death (annexin V). This method offers a reliable way to evaluate immune responses in clinical settings.
Area of Science:
- Immunology
- Cell Biology
- Clinical Research
Background:
- Accurate measurement of cell-mediated cytotoxicity is crucial for clinical research.
- Defining surrogate immunologic endpoints that correlate with clinical outcomes is essential.
Purpose of the Study:
- To develop a novel assay for simultaneous evaluation of target cell death and effector cell frequency.
- To correlate effector cell degranulation, granzyme B secretion, and target cell death.
Main Methods:
- Combined measurement of CD107a expression (effector cell degranulation) and annexin V binding (target cell apoptosis).
- Utilized human cytotoxic T lymphocytes and various effector/target cell systems.
- Employed Granzyme B ELISPOT and flow cytometry assays.
Main Results:
- Demonstrated an incubation time-dependent increase in CD107a expression and annexin V binding.
- Showed excellent correlation between CD107a expression, annexin V binding, and granzyme B secretion.
- Confirmed the specificity and reliability of the developed assay.
Conclusions:
- The novel assay enables simultaneous measurement of cytolytic cell activation, frequency, and target cell death.
- This method provides a valuable tool for assessing immune responses in clinical settings.
- The assay's correlated parameters offer a comprehensive evaluation of cell-mediated cytotoxicity.