Cytokines and PAF release from human monocytes and macrophages: effect of hemoglobin and contaminants.
Monocytes [M] were isolated from venous blood of healthy volunteers and activated macrophage-leukocytes (Mø-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 polymers with M.W. < 400 kDa (LMWHb), [C] Hb crosslinked to form large polymers (< 1,020 kDa) (HMWHb), and Mø-L additionally with [D] UHb contaminated with endotoxin (Hb+E) (2.5 EU/mL), and [E] UHb contaminated with phospholipids (Hb+PLs). The Mø-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 for TNF alpha and GM-CSF at 12, 24 and 36 hours. Mø-L were found more responsive than M colonies. The strongest reaction of Mø-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 much 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 cultures after 36 hours of incubation.
Simoni
J
J
Texas Tech University Health Sciences Center, Department of Surgery, Lubbock, 79430.
Simoni
G
G
Lox
C D
CD
McGunegle
D E
DE
Feola
M
M
D000818:Animals; D001802:Blood Substitutes / Q000302:isolation & purification / Q000633:toxicity; D002417:Cattle; D003432:Cross-Linking Reagents; D016207:Cytokines / Q000378:metabolism*; D004340:Drug Contamination; D004715:Endometriosis / Q000503:physiopathology; D004731:Endotoxins / Q000633:toxicity; D005260:Female; D006454:Hemoglobins / Q000302:isolation & purification / Q000633:toxicity*; D006801:Humans; D066298:In Vitro Techniques; D008055:Lipids / Q000633:toxicity; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D008854:Microscopy, Electron; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D010972:Platelet Activating Factor / Q000378:metabolism*; D011108:Polymers / Q000302:isolation & purification / Q000633:toxicity
D001802:Blood Substitutes; D003432:Cross-Linking Reagents; D016207:Cytokines; D004731:Endotoxins; D006454:Hemoglobins; D008055:Lipids; D010972:Platelet Activating Factor; D011108:Polymers