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. 2017 Dec 7;27:27–39. doi: 10.1016/j.ebiom.2017.12.004

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© 2017 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Fig. 4 — ESAT-6-specific effectors in Mtb memory mice exhibit increased differentiation within the lung parenchyma early during infection compared to H56 memory mice.

A) KLRG1 expression among lung localized I-A^b:ESAT-6-specific CD4 T cells. Representative FACS plots showing KLRG1 expression on ESAT-6_4-17 tet + cells from Mtb (upper plot) and H56 (lower plot) memory mice infected for ten weeks with Mycobacterium tuberculosis relative to their localization in the lung vasculature (CD45.2 IV + ve) or lung parenchyma (CD45.2 IV –ve). Numbers in parentheses represent the percentage of cells expressing KLRG1 in the IV + ve (Red) and the IV − ve population (Blue). Graphs show the proportion of KLRG1 + of I-A^b:ESAT-6_4-17 tetramer + CD4 T cells in the lung vasculature (IV + ve; upper graph) and in the lung parenchyma (IV –ve, lower graph) over the course of infection (unperfused lungs). Mean ± s.d. of 3–4 mice per group at any given time-point. Two-way ANOVA with Sidak's multiple comparison test for simple row effects. Differences between Mtb and H56 memory. ****P < 0.0001, ***P < 0.001, **P < 0.01.

B) PD-1 expression among lung localized I-A^b:ESAT-6-specific CD4 T cells. Representative FACS plots showing PD-1 expression on ESAT-6_4–17 tet + cells from Mtb (upper plot) and H56 (lower plot) memory mice infected for ten weeks with Mycobacterium tuberculosis relative to their localization in the lung vasculature (CD45.2 IV + ve) or lung parenchyma (CD45.2 IV –ve). Numbers in parentheses represent the percentage of cells expressing PD-1 in the IV + ve (Red) and the IV − ve population (Blue). Graphs show the proportion of PD-1 + of I-A^b:ESAT-6_4-17 tetramer + CD4 T cells in the lung vasculature (IV + ve; upper graph) and in the lung parenchyma (IV –ve, lower graph) over the course of infection (unperfused lungs). Mean ± s.d. of 3–4 mice per group at any given time-point. Two-way ANOVA with Sidak's multiple comparison test for simple row effects (ns).

C) ICOS expression among lung localized I-A^b:ESAT-6-specific CD4 T cells. Representative FACS plots showing ICOS expression on ESAT-6_4-17 tet + cells from Mtb (upper plot) and H56 (lower plot) memory mice infected for ten weeks with Mycobacterium tuberculosis relative to their localization in the lung vasculature (CD45.2 IV + ve) or lung parenchyma (CD45.2 IV –ve). Numbers in parentheses represent the percentage of cells expressing ICOS in the IV + ve (Red) and the IV –ve population (Blue). Graphs show the proportion of ICOS + of I-A^b:ESAT-6_4–17 tetramer + CD4 T cells in the lung vasculature (IV + ve; upper graph) and in the lung parenchyma (IV –ve, lower graph) over the course of infection (unperfused lungs). Mean ± s.d. of 3–4 mice per group at any given time-point. Two-way ANOVA with Sidak's multiple comparison test for simple row effects. Differences between Mtb and H56 memory. **P < 0.01.

Gating as depicted in Supplementary Fig. 4.

One of two comparable experiments shown.