Figure 5.

Intracellular TLR2 localization and effect of Bafilomycin A1 treatment in HCMV sensing by NK cells. PBMC samples were analyzed by three color flow cytometry with mAb specific for TLR2 or a control IgG in combination with anti-CD3-PerCP and anti-CD56-APC. For intracellular staining PBMC were fixed and permeabilized prior to the incubation with mAbs. Dot plots display the staining for TLR2 in the NK cell gate (CD56+CD3-) and in monocytes gated by forward and side light scatter. Data correspond to one representative donor out of three analyzed (A). IFNγ secretion was measured by ELISA in supernatants from NK cells incubated with IL-12 and infective- or UV-inactivated HCMV, in the presence or absence of Bafilomycin A1 (Bfa A1, 25 nM). Empty and gray bars correspond to non-treated (NT) and Bfa A1 treated cultures. Bar graph showing the mean ± SEM of the data obtained from six donors (B). Statistic analysis by the Student t-test (*p < 0.05; **p < 0.01; ***p < 0.0001).