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[Preprint]. 2023 Sep 5:2023.09.01.555962. [Version 1] doi: 10.1101/2023.09.01.555962

Fig. 2. In vitro model of NK cell alloreactivity shows variability is dependent on donor ligand and recipient receptor differences.

Fig. 2.

a Cryopreserved PBMC were recovered overnight in RPMI medium supplemented with 10 ng/mL rhIL-15 and 10% heat-inactivated FBS, and then stimulated with K562 wildtype (WT), K562 HLA-E+ and four allogeneic B cell lines expanded with CD40L-expressing fibroblasts and IL-4 at 3:1 E/T. b Heatmap showing MFI of NK cell ligands on stimulator cells; NK cell cognate receptor for ligands are indicated in parentheses. c U-MAP of CD107a expression in the coculture conditions PBMC only and PBMC co-cultured with wildtype K562. d Percentage of CD107a+ NK cells across healthy donors (n=20) when stimulated four allogeneic B cell stimulators, K562 WT and K562 HLA-E+. There were no differences between Stimulators 1–4 except between Stimulator 1 and Stimulator 3. All Stimulators induced less activation than K562 WT and K562 HLA-E+, and there was no difference between K562 WT and K562 HLA-E+. P value was calculated using 2-sided paired Student’s t-test and values were adjusted for multiple testing with Bonferroni correction.