Journal of Immunology Research / 2015 / Article / Fig 1

Research Article

Uptake of CCR7 by KIR2DS4+ NK Cells Is Induced upon Recognition of Certain HLA-C Alleles

Figure 1

De novo expression of CCR7 in NK cell clones upon interaction with transfected 221 cell lines: comparison between KIR2DS4+ and KIR2DS4 NK cell populations. (a) Expression profile of HLA-C and HLA-E molecules (left panels) and of CCR7 receptor (right panel) on 221 cells or 221 cells transfected with HLA-Cw3 or HLA-Cw4 or HLA-Cw6 is shown. (b) KIR2DS4 NKG2A+ or KIR2DS4+ NKG2A+ NK cell clones were incubated with untransfected 221 cells or 221 cells transfected with HLA-Cw3 or HLA-Cw4 or HLA-Cw6 alleles. NK cell clones (6 for each type) were derived from one B haplotype donor. Coincubation was performed in the absence or presence of anti-NKG2A mAbs, as indicated. The histogram represents the percentage of CCR7+ NK cells after incubation with untransfected or HLA-transfected 221 cells. The average of 6 independent experiments and standard deviation (mean ± SD) are indicated. ; . (c) KIR2DS4 NKG2A+ or KIR2DS4+ NKG2A+ NK cell clones (from one B haplotype donor) were incubated with 221-Cw4 cells. 3 KIR2DS4 NKG2A+ and 3 KIR2DS4+ NKG2A+ NK cell clones characterized by low NKG2A-mediated inhibition (on the left) and 3 KIR2DS4 NKG2A+ and 3 KIR2DS4+ NKG2A+ characterized by high NKG2A-mediated inhibition (on the right) are represented. The average of 6 independent experiments and standard deviation (mean ± SD) are indicated. ; . (d) A representative KIR2DS4+ NKG2A+ NK cell clone (derived from one B haplotype donor) was incubated at 37°C with CCR7+ untransfected 221 cells or 221 cells transfected with HLA-Cw4. After 30 minutes, NK cells were harvested and stained with anti-CD56 and anti-CCR7 mAbs for cytofluorimetric analysis. The percentages of CCR7+ cells are indicated in the upper-right corners. (e) KIR2DS4+ NKG2A+ NK cell clones derived from the one B haplotype donor were incubated with untransfected 221 cells or 221 cells transfected with HLA-Cw3 or HLA-Cw4 or HLA-Cw6 alleles. Coincubation was performed in the absence or in the presence of mAbs to the indicated molecules. The histogram represents the percentage of CCR7+ NK cells after incubation with untransfected or HLA-transfected 221 cells. The average of 6 independent experiments and standard deviation (mean ± SD) are indicated. . (f) KIR2DS4+ NKG2A+ NK cell clones derived from one A haplotype donor (upper panel) and one B haplotype donor (lower panel) (3 for each haplotype) were incubated with untransfected 221 cells or 221 cells transfected with HLA-Cw3 or HLA-Cw4 or HLA-Cw6 alleles. The average of 3 independent experiments and standard deviation (mean ± SD) are indicated. .
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