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. 2021 Apr 12;12:2173. doi: 10.1038/s41467-021-22359-x

Fig. 5. The functional response of KIR2DL2 and KIR2DL3 NK cells in the presence of HLA ligands.

Fig. 5

Enriched NK cells from healthy donors were cultured with target cells and the degranulation response of NK cells expressing KIR2D assessed by flow cytometry. a The expression of HLA-C on both 721.221 and Raji, Ramos and Namalwa lymphoma cell lines following staining with DT9 antibody (shaded grey area) is shown, with secondary antibody alone shown as unfilled histogram. b Gating strategy for KIR2DL1/S1, KIR2DL2/S2 and KIR2DL3 NK cells. c Representative flow cytometry analysis of CD107a expression on NK cell subsets from one donor in the presence of 721.221, Raji, Ramos and Namalwa cells is shown (gated on KIR3DL1− NK cells). d Pooled data showing CD107a+ cells from six donors with six KIR2DL1/S1, and KIR2DL3 populations and four KIR2DL2/S2 populations is shown. Each symbol represents an individual population of cells and bars depict mean ± SEM values. CD107a response is normalised to 721.221 response from the same experiment. e Representative CD107a expression on NK cell subsets from one donor in the presence of either 721.221 cells or 721.221 cells transfected HLA-C*06:02 or -C*07:02. f The degranulation response of KIR2D+ NK cells (GL183+ gated on a set GL183 MFI) to untransfected and HLA-C transfected 721.221 cells. Data are pooled from five donors. Each symbol represents data from an individual donor and bars depict mean ± SEM values. CD107a response is normalised to 721.221 response. g The expression of different HLA-C on target cells stained with W6/32 antibody (shaded grey area) is shown with 721.221 stain shown as black line. Statistical significance between KIR3DL2+ and KIR2DL3+ subsets (excluding KIR2DL1+) was tested using Mann–Whitney test for all conditions; no symbol = p > 0.05; *p < 0.05; **p < 0.01. Each experiment has been performed twice with data from both experiments pooled for the figure.