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Updated: Dec 30, 2025
Measurement of Natural Killer Cell-Mediated Cytotoxicity and Migration in the Context of Hepatic Tumor Cells
Published on: February 22, 2020
Two-dimensional dynamic evaluation of natural killer cell-mediated lysis of adherent target cells
Alexandra Frazao1, Louise Rethacker1, Anne Caignard1
1INSERM, U1160, Institut de Recherche Saint-Louis, Hôpital Saint-Louis, Paris, France.
Abstract:
Cell-mediated cytotoxicity is a major function of cytotoxic lymphocytes. The cytotoxic function of immune effectors requires accurate and sensitive quantification as tumor immunotherapies are actively developed to treat growing types of tumors. Various methods have been developed to quantify this function. A first approach consists in measuring the externalization of LAMP-1 (lysosomal associated membrane 1), CD107a molecules transitory expressed at the cell surface by degranulating cytotoxic cells and is determined by flow cytometry. The focus of the chapter concerns the second approach that quantifies target cell lysis resulting from the close contact interaction with cytotoxic Natural Killer (NK) lymphocytes. For long time, target cell lysis was evaluated by chromium release assay, requiring use of radioactive labeled salt (51Cr) and specific devices not compatible with repeated tests performed for immunomonitoring of patients. Other methods include fluorimetric and bioluminescence assays. Monitoring immune cell-mediated lysis of adherent targets by the dynamic measure of cell proliferation using the Real time cell analyzer (RTCA) is a good alternative. The test relies on sensitive dynamic measure of cell index values during their interactions with cytotoxic immune effectors. The cell index increasing with cell proliferation rapidly drops in presence of immune cells. The lysis is quantified in reference with targets alone and expressed as percentages of lysis curves. Moreover, the dynamic monitoring of target cell index allows the evaluation of drugs, cytokines, mAbs on target cell lysis. Here we describe a robust and sensitive method for quantification of immune cell-mediated lysis of adherent targets.
Insights
Quantifying cell-mediated cytotoxicity is crucial for developing tumor immunotherapies. This study presents a robust method using Real-Time Cell Analysis (RTCA) to measure immune cell-mediated target cell lysis, offering a sensitive alternative to radioactive assays.
Area of Science:
- Immunology
- Cell Biology
- Cancer Therapy
Background:
- Cell-mediated cytotoxicity is a key function of cytotoxic lymphocytes, essential for tumor immunotherapy development.
- Accurate quantification of cytotoxic function is vital for monitoring treatment efficacy.
- Existing methods like chromium release assays have limitations, including radioactivity and incompatibility with repeated testing.
Purpose of the Study:
- To describe a robust and sensitive method for quantifying immune cell-mediated lysis of adherent targets.
- To present Real-Time Cell Analysis (RTCA) as an alternative to traditional cytotoxicity assays.
- To highlight the dynamic monitoring capabilities of RTCA for evaluating therapeutic interventions.
Main Methods:
- Utilizes Real-Time Cell Analysis (RTCA) to dynamically monitor target cell proliferation.
- Measures the decrease in cell index values upon interaction with cytotoxic immune effectors (e.g., Natural Killer cells).
- Quantifies target cell lysis by comparing lysis curves to controls (targets alone), expressed as percentages.
Main Results:
- RTCA provides a sensitive and dynamic measure of immune cell-mediated lysis of adherent targets.
- The method allows for the evaluation of drugs, cytokines, and monoclonal antibodies (mAbs) on target cell lysis.
- Demonstrates a robust alternative to radioactive chromium release assays for cytotoxicity assessment.
Conclusions:
- Real-Time Cell Analysis (RTCA) offers a sensitive, non-radioactive method for quantifying immune cell-mediated cytotoxicity.
- This dynamic monitoring approach is valuable for immunomonitoring and evaluating therapeutic agents in cancer immunotherapy.
- The described method provides a robust platform for assessing the efficacy of cytotoxic immune responses against tumors.
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