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. 2020 Oct 30;10:18703. doi: 10.1038/s41598-020-75734-x

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© Crown 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Increased PD-1 expression associates with cytokine⁺ T cells from the lung parenchyma. Groups of mice were immunised as per Fig. 1 and cytokine⁺ CD4 and CD8 T cells derived from the lung parenchyma and lung vasculature compartments detected as described in Figs. 2 and 4. Cells were simultaneously stained for surface expression of PD-1. Representative flow cytometry (A) contoured bivariate plot and (B) histogram both showing the differential expression of PD-1 by cytokine⁺ CD4 T cells derived from the vascular (CD45⁺, grey filled line) and parenchymal (CD45^-, unfilled line) compartments of the lungs of mice receiving the airway BCG/Ad regimen. (C) Graphs showing the median fluorescence intensity (MFI) of PD-1 staining of cytokine⁺ CD4 and CD8 T cells derived from the two lung compartments of mice receiving this vaccine regimen. Circles represent individual animals (n = 11). ***p < 0.001 ****p < 0.0001, two-tailed paired t-test. Data representative of one experiment.