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. 2019 Nov 12;34(3):895–908. doi: 10.1038/s41375-019-0625-3

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© The Author(s) 2019

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — Therapeutic Treg dampen inflammation and support tissue regeneration in the GI tract. CB6F1 recipients were conditioned and transplanted with either BM cells alone (BM control; n = 12–24/time point) or with additional splenocytes (GvHD; n = 10–17/time point) as detailed in Fig. 2. On day 11 after BMT, part of the GvHD animals received donor-derived in vitro expanded Treg cells (Therapy; n = 10–20/time point). At the indicated time points mice were sacrificed and small intestine (SI) and colon were analyzed histopathologically, by FACS and by qRT-PCR. a Absolute numbers of CD45⁺ leukocytes in SI lamina propria (LP) and epithelium (EP). Presence and function of Paneth cells in the SI: b Paneth cells/high power field (HPF, magnification 40×, BM control: n = 6–7/time point; GvHD: 5–15/time point; therapy: n = 8–13/time point) and c lysozyme staining of representative SI specimens (magnification 40×). d Absolute cell numbers of leukocytes in colon LP and epithelium (BM control: n = 12–24/time point; GvHD: 10–17/time point; therapy: n = 10–20/time point) and e histopathological score of the colon at indicated time points post BMT (GvHD day 40: n = 12; therapy day 40: n = 9; therapy day 100: n = 12; BM control day 100: n = 7). f Representative histology after Treg therapy showing mostly normal colon architecture and an area of leukocyte infiltrates (left panel, H/E staining, magnification 200×) and the high frequency of Treg (dark brown/black) in such colonic infiltrates (right panel, staining for Foxp3, magnification 400×). Absolute cell numbers of indicated leukocyte subpopulations, Treg/Tconv ratios and TNF and IFN-γ expression (normalized to levels in nontransplanted animals) in the colon of aGvHD and Treg therapy animals at day 40 after BMT (g) and of Treg therapy and BM control animals at day 100 after BMT (h) (n = 3 for cytokines day 40; n = 4–8 for all other analyses on day 40 and day 100). Combined data from 1 (cytokines day 40) or 3–4 independent experiments are shown. Neutrophils are defined as CD45⁺CD11b⁺Gr-1⁺, DC as CD45⁺CD11c⁺. Proliferating CD4⁺TCRαβ⁺Foxp3⁻ Tconv are identified by Ki-67 expression. Gray-shaded areas indicate respective levels in nontransplanted CB6F1 mice. Summarized data are shown as mean ± s.e.m, (*p < 0.05, **p < 0.01, ***p < 0.001)