Publication: A novel EGFP-CEM-NK<sup>r</sup>flow cytometric method for measuring antibody dependent cell mediated-cytotoxicity (ADCC) activity in HIV-1 infected individuals
Issued Date
2006-08-31
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ISSN
00221759
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2-s2.0-33748772813
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Immunological Methods. Vol.315, No.1-2 (2006), 1-10
Suggested Citation
Wannee Kantakamalakul, Kovit Pattanapanyasat, Surat Jongrakthaitae, Vatcharain Assawadarachai, Silawun Ampol, Ruengpung Sutthent A novel EGFP-CEM-NK<sup>r</sup>flow cytometric method for measuring antibody dependent cell mediated-cytotoxicity (ADCC) activity in HIV-1 infected individuals. Journal of Immunological Methods. Vol.315, No.1-2 (2006), 1-10. doi:10.1016/j.jim.2006.06.005 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/23314
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Title
A novel EGFP-CEM-NK<sup>r</sup>flow cytometric method for measuring antibody dependent cell mediated-cytotoxicity (ADCC) activity in HIV-1 infected individuals
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Abstract
Enhanced green fluorescent protein (EGFP) was stably expressed in CEM-NKrcell, a natural killer (NK) resistant human T-lymphoblastoid cell line, as EGFP-CEM-NKrcells. 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-NKrtarget 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-NKrstable 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. © 2006 Elsevier B.V. All rights reserved.
