IgGs and complement bound to nHZ play a subsidiary role in the immediate nHZ-elicited release of ROS, TNF, and MCP-1 from monocytes. (A) Suspended monocytes from healthy donors were fed with serum-opsonized nHZ (nHZ ops), nonopsonized nHZ (nHZ not ops), or nHZ opsonized with decomplemented serum (nHZ w/o C). Cells were fed immediately after isolation at time 0, briefly spun down, resuspended, and incubated at 37°C. nHZ-elicited burst was quantified 4 minutes after nHZ addition by luminol-enhanced luminescence. (B) Monocytes from healthy donors were enriched by Ficoll passage and adhesion, maintained in culture overnight with 200 U/mL IFNγ, and then supplemented with differently treated nHZ at equal heme content (100nmol/106 monocytes) after 15 hours from isolation. Monocytes were supplemented with opsonized nHZ (nHZ ops), nonopsonized nHZ (nHZ not ops), nHZ opsonized with decomplemented serum (nHZ w/o C), or nHZ opsonized and treated with anti IgG-Fc Abs (nHZ αFc). After 3 hours, supernatants were collected and analyzed for TNF and MCP-1 by ELISA. Four, 5, and 7 independent experiments were performed for TNF, MCP-1, and ROS analysis, respectively. Results are expressed as medians with 95% CI; *P < .05 and #P < .01. Absolute values of nHZ-induced TNF, MCP-1, and ROS release were, respectively, 110 pg/mL (88-122 pg/mL), 430 pg/mL (200-877 pg/mL), and 146 × 103 cps/150 000 cells (74-369 × 103 cps/150 000 cells).
