Figure 10

Exposure to norepinephrine (NE) in vitro impairs T-cell cytokine production in response to PMA/ionomycin stimulation. Enriched splenic T-cells (10⁶ cells/ml) were cultured with 10 μM NE or its vehicle (Vehicle) in vitro. After two days of NE exposure, cells were stimulated with PMA/ionomycin for four hours in the presence of brefeldin A. Intracellular cytokine staining and flow cytometry analysis were performed to measure cytokine production. (A) Representative dot plots show the percentage of IFN-γ⁺ cells and TNF-α⁺ cells in gated CD4⁺ T-cells following PMA/ionomycin stimulation or with brefeldin A only (unstimulated). (B) Bar graph represents the mean ± SEM percentages cytokine producing CD4⁺ T-cell in response to PMA/ionomycin stimulation. (C) Representative dot plots show the percentage of IFN-γ⁺ cells and TNF-α⁺ cells in gated CD8⁺ T-cells following PMA/ionomycin stimulation or with brefeldin A only (unstimulated). (D) Bar graph represents the mean ± SEM percentages of cytokine producing CD8⁺ T-cells in response to PMA/ionomycin stimulation. Ten thousand events gated on lymphocytes were collected. Experiments were performed in triplicate, *P < 0.05, **P < 0.01, one-tailed Student’s t-test.