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. 2018 Feb 28;217(12):1997–2007. doi: 10.1093/infdis/jiy096

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© The Author(s) 2018. Published by Oxford University Press for the Infectious Diseases Society of America.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Interleukin (IL)-7 supports the accumulation of antiviral cytomegalovirus (CMV)-specific T cells. (A) Freshly derived peripheral blood mononuclear cells (PBMCs) from CMV⁺ donors were analyzed for inflammatory cytokine release after CMV-lysate intracellular cytokine staining (ICS) assay after a 7-day culture in the absence (d7, control medium [med], dashed black line) or presence of human recombinant IL-7 (d7, IL-7, black line). Left and right dot plots show the levels of IL-2 and tumor necrosis factor (TNF)α or interferon (IFN)γ and TNFα in gated CD4 T cells, respectively. Background levels of cytokine secretion were typically measured in unstimulated controls (nil). (B) Freshly isolated PBMCs from CMV⁺ donors were rested for 5 days in plain medium (dashed black line) before a 7-day culture in the absence (d12, med, dashed black line) or presence of human recombinant IL-7 (d12, IL-7, black line). At day 12 (d12), CD4⁺ T cells were analyzed for IFNγ and TNFα release (top row) alongside expression of CD127 (bottom row), after CMV-lysate ICS assay. Cytomegalovirus-specific IFNγ⁺ CD4⁺ T cells show high CD127 expression in the resting cultures, whereas they downregulated CD127 expression upon exposure to IL-7. Background levels of cytokine secretion were typically measured in ICS unstimulated (nil) controls. A representative of >10 independent experiments is shown. (C) After subtraction of individual background levels of IFNγ⁺ and TNFα⁺ and IFNγ⁺TNFα⁺ CD4 T cells detected in unstimulated controls (nil), the percentage of CMV-specific cytokine⁺ (IFNγ⁺ and TNFα⁺) CD4 T cells was evaluated in 10 independent CMV⁺ donors in IL-7 (IL-7, open triangles) compared with control medium ([med], open circles) cultures. The graph shows a statistically significant (paired Wilcoxon test, *P = .03) increase of the frequency (Log₁₀) of CMV-specific CD4 T cells after IL-7 culture. (D) CD127 expression (Log₁₀ mean fluorescence intensity [MFI]) is significantly downregulated in CMV-specific IFNγ⁺CD4⁺ T cells exposed to IL-7 (IL-7) compared with control medium (med) at d12 in 6 biologically independent replicates (paired Wilcoxon test, *P = .03).