Ethanol-Induced Alterations of T Cells and Cytokines after Surgery in a Murine Infection Model

Figure 6

Flow cytometric analysis of indicated T helper cell subsets in the lung, relative gene expression of key transcription factors, and release of cytokine protein in a murine model of ethanol exposition for 2 weeks, surgery, and pulmonary infection. (a) Flow cytometric analysis of CD4⁺/IL-17⁺ cells (left) and CD4⁺/Foxp3⁺ cells (right) in the lung of ethanol-treated (ETOH) and saline-treated controls (NaCl) at 24 hours or 48 hours after pulmonary infection with Klebsiella pneumoniae. Cell counts are given as number of cells per 50.000 lymphocytes. (b) Transcriptional regulation of indicated transcription factors assessed by real-time polymerase chain reaction. Expression of β-actin served as housekeeping gene; expression of controls was set to 100%. (c) Protein levels of indicated cytokines were analyzed by cytometric bead array (CBA) and given as μg/mg protein of lung tissue. when comparing ethanol treatment and saline-treated controls at the same time-point (ethanol-dependent effects); ; ; when compared to time- and treatment-matched control (infection-dependent effects); ; when comparing 24 hours to 48 hours (time-dependent effects); data are presented as mean ± SEM, Mann–Whitney U test ( each group). n.s.: not significant; CD: cluster of differentiation; RORγT: retinoid orphan receptor gamma T transcription factor; foxp3: forkhead box p3 T-cell-specific transcription factor; IL: interleukin; ETOH: ethanol treatment; K. pneumoniae: pulmonary infection with Klebsiella pneumoniae.

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