Abstract
Enhanced green fluorescent protein (EGFP) was stably expressed in CEM-NK r cell, a natural killer (NK) resistant human T-lymphoblastoid cell line, as EGFP-CEM-NK r cells. The cells pulsed with HIV-1 gp120 were then used as target cells for the measurement of antibody dependent cell mediated-cytotoxicity (ADCC) by flow cytometry. Compromised EGFP-CEM-NK r target cells stained with propidium iodide (PI) showed dual (green–red) fluorescent. Kinetic studies demonstrated that the sum of ADCC activity measured at 1-h and again at 2-h incubations by this flow cytometric method was comparable to the activity at 6 h by the standard chromium ( 51Cr) release assay (CRA). ADCC activity of HIV-1 seropositive sera measured by this new technique correlated strongly with that of CRA (Pearson's correlation coefficient of 0.832; p-value < 0.001 and intraclass correlation coefficient of 0.903; p-value < 0.001). The EGFP-CEM-NK r stable cell line provides a novel method to measure ADCC activity to HIV-1 gp120 by flow cytometry without pre-staining or pre-labeling target cells.
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