A GFP-transfected HFWT cell line, GHINK-1, as a novel target for non-RI activated natural killer cytotoxicity assay.
An anchorage-dependent Wilms' tumor cell line, HFWT, has been found to be extremely susceptible to human natural killer (NK) cells. Here we established a transfectant of HFWT with the green fluorescence protein (GFP) gene, designated GHINK-1 cells, to apply to the activated NK cytotoxicity assay without radioisotope labeling. After being co-cultured with CD3 CD56+ NK cells, GHINK-1 cells released GFP into the medium. The intensity of the fluorescence from the released GFP correlated almost exactly with the radioactivity of a standard 51Cr-release assay performed with suspension-cultured K562 cells. Because it does not require separation of the remaining live target cells by centrifugation, the non-radioisotopic GFP release assay with GHINK-1 cells is a convenient alternative for monitoring human activated NK killing activity.
Harada
Hideki
H
RIKEN Cell Bank, RIKEN (The Institute of Physical and Chemical Research), Koyadai 3-1-1, Tsukuba Science City, 305-0074, Japan.
Saijo
Kaoru
K
Ishiwata
Isamu
I
Ohno
Tadao
T
D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic; D049452:Green Fluorescent Proteins; D006801:Humans; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology; D007694:Killer Cells, Natural / Q000276:immunology*; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D009396:Wilms Tumor / Q000276:immunology* / Q000378:metabolism
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins