Fig. 5
Dopamine increases IL-1β and inflammasome expression in HIV-infected primary macrophages. A Representative immunofluorescent images at 20X objective (DAPI, blue; Phalloidin, red; p24, green) show human monocyte-derived macrophages (hMDM) infected with 0.5 ng/mL HIVADA and treated with dopamine (10–6 M) for 7 days. Higher levels of cell fusion and giant cell formation as well as p24 expression (white arrows) are seen in infected dopamine-treated cultures relative to cultures only infected with HIV. hMDM were also infected with 0.5 ng/mL HIVADA for 7 days and then treated with dopamine (10–6 M) for 3 h and examined for (B) IL-1β, (C) NLRP3, (D) NLRC4, and (E) AIM2 expression by qPCR. There was a significant increase in IL-1β, NLRP3, and NLRC4 and a trending increase in AIM2 expression with HIV + Dopamine relative to HIV alone. With these same samples we simultaneously extracted protein and examined (F) NLRP3, (G) NLRC4, and (H) AIM2 expression by Western Blot. There was only a significant increase in NLRP3 expression with HIV + Dopamine relative to HIV alone. Significance was determined by paired t-tests and Wilcoxon tests, *p < 0.05 and **p < 0.01