Skip to Main content Skip to Navigation
Book sections

Two-dimensional dynamic evaluation of natural killer cell-mediated lysis of adherent target cells

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.
Complete list of metadata

Cited literature [44 references]  Display  Hide  Download
Contributor : anne caignard Connect in order to contact the contributor
Submitted on : Tuesday, January 14, 2020 - 10:54:38 AM
Last modification on : Saturday, June 5, 2021 - 3:05:16 AM
Long-term archiving on: : Wednesday, April 15, 2020 - 3:24:17 PM


 Restricted access
To satisfy the distribution rights of the publisher, the document is embargoed until : jamais

Please log in to resquest access to the document




Alexandra Frazao, Louise Rethacker, Anne Caignard. Two-dimensional dynamic evaluation of natural killer cell-mediated lysis of adherent target cells. Methods in Enzymology, 631 (ch5), pp.289-304, 2020, ⟨10.1016/bs.mie.2019.07.033⟩. ⟨inserm-02438314⟩



Record views