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/.
Bone marrow effector CD4+ T-cell hematopoiesis. (a–c) Marrow Transcription Factor Expression Assays: 12 week-old male SPF & GF mice were euthanized; femoral bone marrow cells were isolated and stained (n = 4/gp) for flow cytometric analysis to assess (a) % CD4+FOXP3+ (TREG) cells, (b) % CD4+RORγt+ (TH17) cells, and (c) % CD4+T-bet+ (TH1) cells. Percentages are expressed relative to CD4+ cells. (d–g) Marrow Intracellular Cytokine Expression Assays: 11 week-old gender matched (2 male and 2 female per group) SPF & GF mice were euthanized; femoral whole marrow was plated overnight for cytokine activation (PMA, Ionomycin, Monensin). Cells were isolated and stained (n = 4/gp) for flow cytometric analysis to assess (d) % CD3+CD4+CD8−IL10+IL17a− (CD4+IL10+) cells, (e) % CD3+CD4+CD8−IL10−IL17a+ (CD4+IL17a+) cells, (f) % CD3+CD4+CD8−IFNγ−IL17a+ (CD4+IL17a+) cells, and (g) % CD3+CD4+CD8−IFNγ+IL17a− (CD4+IFNγ+) cells. Percentages are expressed relative to CD3+CD4+CD8− cells. Data are reported as mean ± SEM. *p < 0.05 vs. SPF; **p < 0.01 vs. SPF.