PMID: 11334968May 4, 2001Paper

Development of a novel flow cytometric cell-mediated cytotoxicity assay using the fluorophores PKH-26 and TO-PRO-3 iodide

Journal of Immunological Methods
A E Lee-MacAryR W Wilkinson

Abstract

A flow cytometric (FCM) assay has been developed for the determination of cell-mediated cytotoxicity (CMC). In the assay, the target tumour cell population was labelled with a membrane dye, PKH-26, prior to incubation with splenocyte effector cells. Cell death within the target population was assessed by the addition of the viability probe TO-PRO-3 iodide (TP3) and analysed by flow cytometry. The extent of cytotoxicity was determined by the relative number of live target cells labelled with PKH-26 only and dead, permeabilised cells labelled with both PKH-26 and TP3. This CMC method allows the analysis to be conducted on a single cell basis and overcomes the need for radiochemicals. This communication indicates that the FCM assay is an accurate and reproducible experimental system capable of analysing natural killer (NK) cell and antibody-dependent cell-mediated cytotoxicity. The procedure is comparable to the chromium release assay. We believe that this is one of the first demonstrations of an FCM-based antibody-dependent cell-mediated cytotoxicity (ADCC) assay.

References

Oct 1, 1965·Experimental Cell Research·H O Sjögren, I Hellström

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Citations

Aug 27, 2013·Archivum Immunologiae Et Therapiae Experimentalis·Marta PokrywczynskaTomasz Drewa
Oct 31, 2001·Journal of Immunological Methods·R W WilkinsonE L Ross
Sep 7, 2002·Journal of Immunological Methods·Yan-Yi Wang, Xiao-Xiang Zheng
Nov 6, 2003·Toxicology·Maritha Marcusson-Ståhl, Karin Cederbrant
Jan 8, 2011·International Immunology·Anna LünemannChristian Münz
Jul 3, 2013·Proceedings of the National Academy of Sciences of the United States of America·Francesco MarrasAndrea De Maria
Sep 26, 2013·Journal of Immunological Methods·Pauline GourdainSylvie Le Gall
Nov 15, 2011·International Immunopharmacology·Ryo KakutaniNaohito Ohno
Mar 30, 2010·Nuclear Medicine and Biology·Joon-Kee YoonYoung Hwan Ahn
May 29, 2007·Journal of Immunological Methods·Marguerite BracherSophia N Karagiannis
Oct 6, 2015·The Journal of Clinical Investigation·Isaak QuastJan D Lünemann
Jan 30, 2004·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Piotr PozarowskiZbigniew Darzynkiewicz
Oct 20, 2006·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Wlodek MandeckiZbigniew Darzynkiewicz
Jul 19, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Carel A van BaalenGuus F Rimmelzwaan
Mar 22, 2005·Journal of Immunological Methods·Michael R OlinThomas W Molitor
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Jun 4, 2010·Expert Review of Vaccines·Liubov ZaritskayaAnatoli M Malyguine
Jan 6, 2009·Annals of the New York Academy of Sciences·Gwen S ScottF Susan Wong
Jun 18, 2010·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Vincent R BonaguraAllan L Abramson
Dec 17, 2014·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Jinyao ChenLishi Zhang
Jun 13, 2012·Journal of Neuroimmunology·Fabian WolpertGünter Eisele
Dec 29, 2007·Immunological Investigations·Paul K Wallace, Katharine A Muirhead
Dec 29, 2007·Immunological Investigations·Joseph D TarioPaul K Wallace
Apr 13, 2007·The Journal of Biological Chemistry·Bhaswati BandyopadhyayJeff Kuret

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