Figure 7
Figure 7. IFNγ production, but not CD107a expression, is increased after CMV reactivation. PBMCs from seronegative (black bars, n = 10), seropositive (no CMV reactivation, white bars, n = 5), and seropositive (CMV reactivation, gray bars, n = 8) recipients at the indicated time points were incubated in either medium alone (not shown) or with K562 cells for 5 hours. (A) CD107a expression (left panel) and IFNγ production (right panel) was measured on NKG2C+ NK cells. Bars represent the means ± SEM. NKG2C+ cells from different groups of recipients were compared at each time point using the Student t test. *P ≤ .05. (B) NKG2C+ NK cells from seropositive (CMV reactivation) recipients were further gated as being KIR+ (black bars) or KIR− (white bars) based on staining with a cocktail of Abs against CD158a/h, CD158b/j, and CD158e, and IFNγ production was measured for each subset. Bars represent the means ± SEM. KIR+ and KIR− NK cells were compared at each time point using the paired t test. *P ≤ .05. (C) NKG2C+ NK cells from seropositive (CMV reactivation) recipients were further gated as being CD57+ (black bars) or CD57− (white bars) and IFNγ production was measured for each subset. Bars represent the means ± SEM. CD57+ and CD57− NK cells were compared at each time point using the paired t test. *P ≤ .05. (D) IFNγ, T-bet, and IL-15Rα mRNA expression were measured on resting PBMCs at 6 months and 1 year after HCT. Samples were normalized to 18S RNA. Bars represent the means ± SEM. Recipients who reactivated CMV and those who did not were compared at each time point using the t test. *P ≤ .05, **P < .01, and ***P < .001.

IFNγ production, but not CD107a expression, is increased after CMV reactivation. PBMCs from seronegative (black bars, n = 10), seropositive (no CMV reactivation, white bars, n = 5), and seropositive (CMV reactivation, gray bars, n = 8) recipients at the indicated time points were incubated in either medium alone (not shown) or with K562 cells for 5 hours. (A) CD107a expression (left panel) and IFNγ production (right panel) was measured on NKG2C+ NK cells. Bars represent the means ± SEM. NKG2C+ cells from different groups of recipients were compared at each time point using the Student t test. *P ≤ .05. (B) NKG2C+ NK cells from seropositive (CMV reactivation) recipients were further gated as being KIR+ (black bars) or KIR (white bars) based on staining with a cocktail of Abs against CD158a/h, CD158b/j, and CD158e, and IFNγ production was measured for each subset. Bars represent the means ± SEM. KIR+ and KIR NK cells were compared at each time point using the paired t test. *P ≤ .05. (C) NKG2C+ NK cells from seropositive (CMV reactivation) recipients were further gated as being CD57+ (black bars) or CD57 (white bars) and IFNγ production was measured for each subset. Bars represent the means ± SEM. CD57+ and CD57 NK cells were compared at each time point using the paired t test. *P ≤ .05. (D) IFNγ, T-bet, and IL-15Rα mRNA expression were measured on resting PBMCs at 6 months and 1 year after HCT. Samples were normalized to 18S RNA. Bars represent the means ± SEM. Recipients who reactivated CMV and those who did not were compared at each time point using the t test. *P ≤ .05, **P < .01, and ***P < .001.

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