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

Fig. 5. Alloreactivity of educated NK cells transcend donor differences and is maintained post-transplant.

Fig. 5.

Cryopreserved pre- and post-transplant PBMC from CTOT01 kidney transplant (n=70) were recovered overnight in 10 ng/mL rhIL-15 and stimulated with donor allo-stimulator B cells for 6 hours at 3:1 E/T. Results were profiled by CyTOF and subsets were defined by gating on CD56dim NK cells followed by combinatorial expression of educating inhibitory receptors, NKG2A, KIR3DL1, KIR3DL2, KIR2DL1 and KIR2DL3 and the CD57 maturation marker. Boxplot shows percent CD107a+ in recipient NK cell subsets in response to co-culture with donor cells for which pre- and post-transplant cells were available (n=34). When recipient NK cells were stimulated with donor cells, the NKG2A+KIR+ educated subset produced the most CD107a while the uneducated NKG2A–KIR− subset produced the least CD107a. The effect of NKG2A/KIR education on CD107a expression in response to donor cells persisted 6+ months post-transplant. P value was calculated as one-way ANOVA as indicated and as 2-sided paired Student’s t-test; t-test p values were adjusted for multiple testing with Bonferroni correction and ns indicates p > 0.1.