CpG ODNs induce modulation of surface KIR3DL2 on IL2-activated NK cell populations and Sézary lymphoma T cells. (A) IL2-activated NK cell populations were cultured for 20 hours either in the absence or in the presence of ODN A (5 μg/mL), ODN B (5 μg/mL), or ODN C (5 μg/mL) and then assessed by cytofluorimetric analysis for the surface expression of various NK cell markers. Black profiles indicate the expression of different NK cell molecules. Gray profiles correspond to isotypic controls. The MFI of the positive peak is indicated in each histogram. Results are representative of 10 distinct experiments. (B) HUT78, PNO, and CS cell lines were cultured for 20 hours either in the absence (gray profiles) or in the presence (black profiles) of ODN C (5 μg/mL) and then analyzed for KIR3DL2 and CD3 surface expression. Results are representative of 4 distinct experiments. (C) Polymerase chain reaction analysis of TLR9 transcript in HUT 78, CS, and PNO cell lines. Plus signs (+) indicate samples obtained by retrotranscription in the presence of RT enzyme, minus signs (−) samples obtained in the absence of RT enzyme, used as control for DNA contaminations. Molecular weight marker (MW, given in bp) and no template control (NTC) are also reported. Results are representative of 3 independent experiments. (D) For KIR3DL2-modulation cytofluorimetric analysis of KIR3DL2, expression was assessed on IL2-activated NK cell populations stimulated with ODN C for different time intervals. For KIR3DL2 re-expression, after 20 hours of stimulation, cells were harvested, washed, and cultured without ODN for different time intervals. Surface re-expression of KIR3DL2 was analyzed at each time point. Black profiles indicate KIR3DL2 expression, whereas gray profiles correspond to isotypic controls. The percentage of positive cells and the MFI of the positive peaks are indicated in each histogram. Results are representative of 5 distinct experiments.