Anti cd16 3g8

The anaplastic thyroid cancer cell line (ARO) was cultured in highglucose Dulbecco modified Eagle medium (Gibco) supplemented with fetal calf serum, 10%; penicillin/streptomycin (penicillin G, 10,000 U, and streptomycin, 10 mg), 10 mL/L; amphotericin B (250 mg/mL), 10 mL/L; and L-glutamine, 1% (16) . NKs were obtained from the blood of healthy donors. Healthy donors' peripheral blood mononuclear cells (4 • 10 5 cells) were isolated by Lymphoprep (Stemcell Technologies) gradient centrifugation and then cocultured for 10 d with irradiated (30 Gy) Epstein-Barr virus-transformed B-cell line RPMI 8866 (10 5 cells) at 37°C as previously described (17, 18) . After 10 d, the cells were collected and phenotypically characterized through immunofluorescence using anti-CD16 (3G8; BD-Biosciences), anti-CD56 (BD-Biosciences), and anti-CD3 (BD-Biosciences) mAb and analyzed by flow cytometry (BD-Biosciences). On day 10 the cell population was routinely greater than 90% CD56 1 CD16 1 CD3 2 ; when purity was less than 90%, contaminant T-cells were eliminated by immunomagnetic negative selection with anti-CD3 mAb to obtain a purity greater than 95%. Human granulocytes were isolated from healthy donors by Percoll (GE Healthcare) gradient centrifugation as previously described (19) .