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. 2008 Feb 8;4(2):e30. doi: 10.1371/journal.ppat.0040030

Figure 1. The Rapid Kinetics of the NK Cell Response Controls the Early Dissemination of ECTV to Central Organs by Using IFN-γ- and Perforin-Dependent Mechanisms.

Figure 1

Intact or NK cell–depleted (treated with anti-NK1.1 mAb PK136) B6 mice were infected with 3,000 pfu ECTV. (A) 7 d PI, the absolute numbers of live lymphocytes in the spleen were determined by trypan blue exclusion. Data correspond to the average ± SD of pooled spleens of three mice from five independent experiments.

(B) 7 d PI, virus titers in spleen and liver were determined by plaque assay. Data correspond to the average ± SD of six individual mice from two independent experiments.

(C) B6 mice were infected with ECTV in the footpad. The NK cell response, as determined by the percentage of NK cells expressing intracellular IFN-γ, at the indicated times PI was determined in the indicated organs. Data correspond to pools of three mice and are representative of three similar experiments.

(D) Representative flow cytometric analysis of D-LN from mice infected for 2 d with ECTV and from control uninfected mice. Upper panel: Dot plots indicating the proportion of NK cells (NK1.1+, CD3ɛ−). Lower panel: GzB and IFN-γ production by gated NK cells (NK1.1+/CD3ɛ− gate of the upper panels). Data correspond to pools of three mice and are representative of at least five experiments. >98% of cells stained with control Ig were in the lower left quadrant of the dot plots (not shown).

(E) B6 mice were infected with ECTV, and NK cell (NK1.1+, CD3ɛ−) proliferation was determined at different times PI in the indicated organs by using a 3-h BrdU incorporation assay. Data are representative of three independent experiments.

(F) Flow cytometric analysis of D-LN from mice infected for 2 d with ECTV and from control uninfected mice. Upper panels: Plots are gated on NK cells (NK1.1+, CD3ɛ−). Lower panels: Gated on R1, R2, and R3 populations from the upper-right (infected) plot. Data are representative of three independent experiments.

(G) Intact B6 mice and B6 mice depleted of NK cells (treated with anti-NK1.1) or T cells (treated with anti-CD4 and anti-CD8) were infected with ECTV and virus titers in spleen and liver were determined on day 3 PI. Data are the average ± SD of six individual mice from two experiments.

(H) Wild-type and IFN-γ-deficient B6 mice were infected with ECTV in the footpad, and virus titers were determined 3 d PI. Data are the average ± SD of three individual mice and is representative of two individual experiments.

(I) Wild-type and Pf-deficient B6 mice were infected with ECTV in the footpad and virus titers were determined 3 d PI. Data are the average ± SD of three individual mice and are representative of two individual experiments.