Figure 3

Chronic alcohol drinking alters immune responses to LPS. (a) Experimental design: PBMC from controls (n = 4), CMD (n = 4), and CHD (n = 4) animals were cultured in the presence or absence of LPS for 16 h. Supernatants were collected for immune mediator measurements using multiplex bead assay and the cell pellets were used for RNA extraction and RNA-Seq. (b) Principal Component Analysis (PCA) of overall immune mediator profiles (21 analytes detected above limits of detection) produced in the absence of LPS stimulation. Each circle represents a biological sample. (c) Heatmap of median secreted analyte concentrations (pg/mL) before and after LPS stimulation. Dose associated linear increases or decreases in secreted levels both before and after stimulation were independently tested using linear regression analysis. Analytes with statistically significant and trending linear association with ethanol consumption dose are marked (t – p = 0.05; *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.001, F-tests). Protein concentrations are scaled from low (light green) to high (dark green).