Sanne Hoeks

Chapter 3 36 Fig 1 Journal of Allergy and Clinical Immunology 2013 132754-756.e3DOI: (10.1016/j.jaci.2013.04.014) FIGURE 1. CB T cells are deficient in T H 17 development but do upregulate FOXP3. APB and CB naïve T cells were activated by plate bound anti-CD3 in the presence of viable APC for 6 days. IL-17 + and FOXP3 + CD4 + T cell numbers were assessed by flow cytometry. A, Representative cytometry plots. B, Supernatant IL-17 concentration (n=8) and percentage of IL-17 + CD4 cells (n=8 APB, n=5 CB). C, Supernatant IL-21, IL-22 and GM-CSF (n=4 APB, n=5 CB). D, Percentage of FOXP3 + CD4 + cells (n=8). *p<0.05, **p<0.01, Mann Whitney U Test. FIGURE E1. A, Culture supernatant IL-17 concentration after culture for 6 days with anti-CD3 and APCs or with APCs replaced by anti-CD28. n = 4 for APB and 5 for CB. B, IL-10, IFN-γ, and IL-13 concentration in APB and CB cell culture supernatants. APB and CB-naive T cells were cultured with APCs and plate- bound anti-CD3 for 6 days. Supernatants were subsequently analyzed for cytokine. n = 4 for APB and 5 for CB. C, Flow cytometric analysis of IL-17 and IL-21 or IL-22 production in APB cells. Naive T cells were cultured with APCs and plate-bound anti-CD3 for 6 days, restimulated with phorbol 12-myristate 13-acetate, ionomycin, and monensin and subsequently analyzed for production of IL-17, IL-21, and IL-22. Representative for n = 5. D, CD161 expression does influence T H 17 development in CB. APB and CB T cells were sorted as CD25 − CD45RO − CD161 − or CD4 + CD161 + and subsequently activated by anti-CD3 in the presence of viable APCs without addition of recombinant cytokines. Left plot, APB CD161 + cells become IL-17 producers rather than APB CD161 − cells. CB cells do not produce IL-17 regardless of their CD161 expression. Right plot, CB T cells upregulate FOXP3 independently of CD161 expression. n = 3. *P < .05 and **P < .01, Mann-Whitney U Test. ▶