High prevalence of NK cells displaying lack of intracellular signaling protein and PLZF expression in patients with GATA2 mutation. PBMCs from CMV-seronegative (n = 69) and -seropositive (n = 127) healthy donors as well as patients (n = 10) and asymptomatic carriers (n = 3) with heterozygous GATA2 mutations were examined by flow cytometry. Graphs depict (A) the number of total NK cells as defined by CD3⁻CD56⁺/CD16⁺ cells in the lymphocyte gate, (B) the percentage of CD3⁻CD56^bright NK cells within the total CD3⁻CD56⁺/CD16⁺ NK-cell population, and (C) the percentage of CD3⁻CD56^dim NK cells expressing CD57. (D-H) Contour plots show CD3⁻CD56^dim NK cells from representative CMV-seronegative and -seropositive healthy donors as well as 2 patients and 1 asymptomatic carrier with GATA2 mutations. Expression of CD57 is plotted against (D) NKG2C or the intracellular proteins (E) FcεRγ, (F) SYK, (G) EAT-2, and (H) PLZF. (D-H) Graphs depict the percentages of CD3⁻CD56^dim NK cells expressing NKG2C, FcεRγ, SYK, EAT-2, or PLZF, as indicated. In the graphs (A-H), CMV serostatus for GATA2 patients and carriers is indicated as seronegative (open circles), seropositive (gray circles), or undetermined (black circles). (A-H) Boxes indicate 25th, 50th (median), and 75th percentiles, with a plus indicating the mean. For box plots, whiskers indicate 95% confidence intervals. Otherwise, error bars indicate mean with standard deviation (SD). P values were determined using the Student unpaired t test. *P < .05, **P < .01, and ***P < .001. (I) The overall frequency of healthy controls (ctrls, stratified on CMV seropositivity, or individuals with GATA2 mutations with significant, sizeable downregulations of 1, 2, or 3 of the signaling proteins FcεRγ, SYK, and EAT-2 is plotted. (J) In individuals with GATA2 mutation, the percentage of CD3⁻CD56^bright NK cells within the total CD3⁻CD56⁺/CD16⁺ NK-cell population is plotted as a function of the frequency of PLZF-expressing CD3⁻CD56^dim NK cells. (K) Among healthy controls, the percentage of CD3⁻CD56^bright NK cells within the total CD3⁻CD56⁺/CD16⁺ NK-cell population is plotted as a function of the frequency of PLZF-expressing CD3⁻CD56^dim NK cells. The CMV serostatus is indicated as seronegative (open circles) seropositive (gray circles). (L) Among healthy controls with sizeable adaptive NK-cell expansions as defined by PLZF deficiency, the percentage of CD3⁻CD56^bright NK cells within the total CD3⁻CD56⁺/CD16⁺ NK-cell population is plotted as a function of the frequency of PLZF-expressing CD3⁻CD56^dim NK cells. Healthy controls with sizeable adaptive NK-cell expansions were overwhelmingly CMV seropositive (gray circles).