Adrenal-Derived Hormones Differentially Modulate Intestinal Immunity in Experimental Colitis

Figure 2

Reduced accumulation of regulatory T cells (Treg) in the absence of adrenal glands. C57BL/6 mice were adrenalectomized and exposed to dextran sulfate sodium (DSS) 3%. The spleen and MLN (mesenteric lymph nodes) were collected on the 6th day of colitis and processed for staining with specific antibodies for regulatory markers and flow cytometry. Analyses were performed according to the cell size (FSC) and granularity (SSC), followed by a selecting gate in the lymphocytes region and evaluation of dot plots for CD4⁺CD25⁺ and CD4⁺CD25⁻ cells (a), in which the frequency of Treg markers was quantified by histograms. (b) Percentage of CD4⁺CD25⁻PD1⁺ and CD4⁺CD25⁺PD1⁺ leukocytes from spleen and MLN. (c) Frequency of CD4⁺CD25⁻CD73⁺ and CD4⁺CD25⁺CD73⁺ cells from spleen and MLN. (d) Percentage of CD4⁺CD25⁻FR4⁺ and CD4⁺CD25⁺FR4⁺ leukocytes from spleen and MLN. (e) Frequency of CD4⁺CD25⁻FoxP3⁺ and CD4⁺CD25⁺FoxP3⁺ cells from spleen and MLN. (f) Mean fluorescence intensity (MFI) of FoxP3 in the CD4⁺CD25⁺ population of spleen and MLN. DSS: C57BL/6 mice exposed to DSS; ADX + DSS: C57BL/6 adrenalectomized mice exposed to DSS. The dotted lines correspond to the control group without colitis. Results are representative of two independent experiments, with 5 mice/group. .

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