Hyper-activated CD16⁺ monocytes preferentially produce Th17-related cytokines.

<p>(A) Histogram plots of HLA-DR expression on monocytes and CD16⁺ monocytes from circulating monocytes (blood) and liver-infiltrating monocytes (liver) in a representative IA individual. (B) The MFI values of HLA-DR expressed on monocytes and CD16⁺ monocytes from CHB patients in each group. Each open dot represents one subject with lower HAI scores and each solid dot represents patients with higher HAI scores. (C) Representative histogram plots of TNF-α, IL-6, IL-1β, IL-12/23 p40 expression (red line histograms) or isotype control (green line histograms) on CD14⁺ CD16⁻ monocytes and CD16⁺ monocytes in an IA subject. (D) Data shown are representative of ten independently performed experiments from IA subjects. (E) Representative expansion of Th17 cells with the cytokine cocktail (IL-6, IL-1β and IL-23) and TCR-stimulation. Values in the upper left quadrant indicate the percentages of CFSE^low Th17 cells. (F) Individual values of the fold changes in the percentage of total and CFSE^low Th17 cells. Horizontal bars indicate the median fold changes. (G) Representative expansion of Th17 cells with activated CD16⁺ monocytes in HC subjects (n = 4). Purified CD4⁺ T cells from HCs were cocultured with autologous CD16⁺ monocytes and anti-CD3/CD28 microbeads in the absence or presence of LPS. After 4 days, cells were stimulated for 6 h with PMA/ionomycin and Golgi-stop (during the last 5 h), and IL-17 expression and production were determined by ICC staining. Values in the upper quadrant represent the percentage of Th17 cells. MFI, mean fluorescence intensity; HAI, histological activity index; IA, immune activated patients; CFSE, Carboxyfluorescein succinimidyl ester.