CYTOKINES AND PAF RELEASE FROM HUMAN MONOCYTES AND MACROPHAGES - EFFECT OF HEMOGLOBIN AND CONTAMINANTS

Monocytes [M] were isolated from venous blood of healthy volunteers an d activated macrophage-leukocytes (M0-L) were obtained from peritoneal fluid of patients with mild endometriosis. The M were incubated with pyrogen free CELLGRO culture medium [Control], and with 0.2 mM of [A] unmodified bovine hemoglobin (UHb), [B] Hb crosslinked to form polymer s with M.W.<400 kDa (LMWHb), [C] Hb crosslinked to form large polymers (<1,020 kDa) (HMWHb), and M0-L additionally with [D] UHb contaminated with endotoxin (Hb+E) (2.5 EU/mL), and [E] UHb, contaminated with pho spholipids (Hb+PLs). The M0-L medium of incubation was tested for TNF alpha, IL-1 alpha, IL-6, GM-CSF and PAF after 6 and 24 hours, but M fo r TNF alpha and GM-CSF at 12, 24 and 36 hours. MT?-L were found more r esponsive than M colonies. The strongest reaction of M0-L was to Hb+E, which produced levels of cytokines and PAF higher than Controls (p<0. 001). Hb+PLs induced smaller increases of TNF and IL-6, and a decrease in the levels of IL-1 and GM-CSF. However, the release of PAF was muc h greater with this Hb than with Hb+E. UHb caused an increase in TNF, as compared to control (p<0.01). LMWHb generated a similar increase in TNF, but also a decrease in IL-1. Both polymerized Hb forms inhibited expression of GM-CSF. HMWHb induced high levels of TNF, IL-1 and PAF. UHb, LMWHb and HMWHb significantly increase levels of TNF in M cultur es after 36 hours of incubation.