期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 272, 期 1-2, 页码 189-197出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0022-1759(02)00505-7
关键词
natural killer cell; cytotoxicity assay; flow cytometry; enhanced green fluorescent protein
Enhanced green fluorescent protein (EGFP) was stably expressed in human erythroleukaemia K562 cells (EGFP-K562) and used as target cells for measurement of natural killer (NK) cell cytotoxicity by flow cytometty. The compromised EGFP-K562 target cells were stained with propidium iodide (PI) and showed dual (green-red) fluorescent. Although the kinetic study demonstrated that the optimal incubation time for the assay was 4 h, a 2-h incubation. period also gave comparable results. This new technique correlated strongly with the standard chromium (Cr-51) release assay at the correlation coefficients of 0.87 and 0.89 at p-value <0.001 for 2- and 4-h incubation times, respectively. The EGFP-K562 stable cell line provides a novel method to measure NK cytotoxicity by flow cytometry without pre-staining or pre-labeling target cells. (C) 2002 Elsevier Science B.V. All rights reserved.
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