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. 2018 Jun 13;9:1344. doi: 10.3389/fimmu.2018.01344

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Copyright © 2018 Littringer, Moresi, Rakebrandt, Zhou, Schorer, Dolowschiak, Kirchner, Rost, Keller, McHugh, LeibundGut-Landmann, Robinson and Joller.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Expression of co-inhibitory receptors and granzymes is a mutual feature of Th1-driven Treg specialization. C57BL/6 mice were acutely infected with LCMV WE (blue), Legionella pneumophila (Lpn, red), Vaccinia Virus (VV, black), C. albicans (yellow), or left naïve and expression of the co-inhibitory receptors Tim-3, CD85k, Lag-3, and TIGIT (A,C), or granzyme B (B,D) among live CD4⁺Foxp3⁺ Treg cells was determined by flow cytometry. (A,B) Peak expression levels of (A) TIGIT, CD85k, Lag-3, Tim-3, and (B) granzyme B among splenic CD4⁺Foxp3⁺ Treg cells (Lpn: day 5–7; VV: day 7–10; LCMV: day 10–14) or among CD4⁺Foxp3⁺ Treg cells isolated from cervical LNs (C. albicans, day 7). (C,D) Frequencies of CD4⁺Foxp3⁺ Treg cells expressing the indicated markers over time or in naïve controls are depicted. Summary data (mean ± SD; biological replicates; LCMV, VV and Lpn n = 5; C.a n = 6; naïve spleen n = 10, naïve LN n = 3) and representative plots of >3 (LCMV, VV) or two (Lpn, C. albicans) independent experiments are shown (*p < 0.05, **p < 0.01, ***p < 0.001) (Mann–Whitney test).

Expression of co-inhibitory receptors and granzymes is a mutual feature of Th1-driven Treg specialization. C57BL/6 mice were acutely infected with LCMV WE (blue), Legionella pneumophila (Lpn, red), Vaccinia Virus (VV, black), C. albicans (yellow), or left naïve and expression of the co-inhibitory receptors Tim-3, CD85k, Lag-3, and TIGIT (A,C), or granzyme B (B,D) among live CD4⁺Foxp3⁺ Treg cells was determined by flow cytometry. (A,B) Peak expression levels of (A) TIGIT, CD85k, Lag-3, Tim-3, and (B) granzyme B among splenic CD4⁺Foxp3⁺ Treg cells (Lpn: day 5–7; VV: day 7–10; LCMV: day 10–14) or among CD4⁺Foxp3⁺ Treg cells isolated from cervical LNs (C. albicans, day 7). (C,D) Frequencies of CD4⁺Foxp3⁺ Treg cells expressing the indicated markers over time or in naïve controls are depicted. Summary data (mean ± SD; biological replicates; LCMV, VV and Lpn n = 5; C.a n = 6; naïve spleen n = 10, naïve LN n = 3) and representative plots of >3 (LCMV, VV) or two (Lpn, C. albicans) independent experiments are shown (*p < 0.05, **p < 0.01, ***p < 0.001) (Mann–Whitney test).