Details

Autor(en) / Beteiligte

• Kantakamalakul, Wannee
• Pattanapanyasat, Kovit
• Jongrakthaitae, Surat
• Assawadarachai, Vatcharain
• Ampol, Silawun
• Sutthent, Ruengpung

Titel

A novel EGFP-CEM-NK r flow cytometric method for measuring antibody dependent cell mediated-cytotoxicity (ADCC) activity in HIV-1 infected individuals

Ist Teil von

• Journal of immunological methods, 2006, Vol.315 (1), p.1-10

Ort / Verlag

Elsevier B.V

Erscheinungsjahr

2006

Link zum Volltext

Quelle

Elsevier ScienceDirect Journals Complete

Beschreibungen/Notizen

• 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.