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. 2015 Mar 9;10(3):e0118685. doi: 10.1371/journal.pone.0118685

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© 2015 Sakala et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 5 — Mice (n = 5–10/group) were infected with ECTV-WT or ECTV-IFN-γbp^Δ, sacrificed on day 7 p.i. and splenocytes used for intracellular staining for IFN-γ, IL-4, T-bet or GATA-3 without stimulation (A and B) or with PMA + ionomycin stimulation (C and D). Shown are means of absolute numbers of IFN-γ⁺CD4 T cells with no stimulation (A) or PMA + ionomycin stimulation (C), IL-4⁺CD4 T cells with no stimulation (B) or PMA + ionomycin stimulation (D). Also shown are means of absolute numbers of T-bet⁺ and GATA-3⁺ CD4 T cells following ECTV-WT (E) or ECTV-IFN-γbp^Δ (F) infection. Two-way ANOVA followed by Fisher’s LSD test for significance was used. For panel A, numbers of IFN-γ⁺CD4 T cells in WT mice for both viruses were significantly higher (p<0.0001) compared to numbers in all GKO strains. In IL-4^−/− and STAT-6^−/− mice, IFN-γ⁺CD4 T cell numbers generated by ECTV-IFN-γbp^Δ infection were significantly higher (p = 0.0005 and p = 0.0013, respectively) compared to WT virus infection. In IL-13^−/− mice, IFN-γ⁺CD4 T cell numbers generated by WT virus infection were significantly higher (p = 0.0034) compared to ECTV-IFN-γbp^Δ infection. For panel B, numbers of IL-4⁺CD4 T cells in WT mice for both viruses were significantly higher (p<0.01) compared to numbers in all GKO strains. No other significant differences were found. For panel C, numbers of IFN-γ⁺CD4 T cells in WT mice for both viruses were significantly higher (p<0.0001 in ECTV-WT and p < 0.001 in ECTV-IFN-γbp^Δ) compared to numbers in all GKO strains. In IL-4^−/−, STAT-6^−/− IL-4Rα^−/− and IL-13^−/− / IL-4Rα^−/− mice, IFN-γ⁺CD4 T cell numbers generated by ECTV-IFN-γbp^Δ infection were significantly higher compared to WT virus infection. For panel D, numbers of IL-4⁺CD4 T cells in WT mice for ECTV-WT virus was significantly higher (p<0.05) compared to numbers in all GKO strains. In WT mice, IL-4⁺CD4 T cell numbers generated by WT virus infection were significantly higher (p<0.05) compared to ECTV-IFN-γbp^Δ infection. For panel E, T-bet⁺ CD4 T cell numbers were significantly increased (p<0.01) in STAT-6^−/− and IL-13^−/− mice compared to WT animals. For panel F, no significant differences were found between strains. Data shown for panels A and B are from one of two independent experiments with similar results. Data shown for panels C-F are from one experiment. *, p < 0.05; **, p < 0.01; ***, p < 0.0001.