Contribution of CR3 to nitric oxide production from macrophages stimulated with high-dose of LPS.
The contribution of the complement receptor type 3 (CR3) to nitric oxide (NO) production from macrophages stimulated by LPS was investigated. When thioglycollate-elicited mouse peritoneal macrophages were stimulated with a high dose of LPS (10 micrograms/ml) in both the presence and absence of fetal calf serum, a source of LPS binding protein (LBP) necessary for the binding of LPS to CD14, NO production was observed. These findings suggest that CD14-dependent and CD14-independent signaling pathways for NO production are present in macrophages. Because binding and phagocytosis of bacteria by macrophages through the CR3 has been previously reported, we investigated whether the CR3 acts in CD14-independent signaling pathway for NO production. By flow cytometric analysis, the binding of FITC-labeled anti-CR3 monoclonal antibody (anti-CR3 mAb) to macrophages was inhibited by LPS. Anti-CR3 mAb induced iNOS protein and produced NO in a dose dependent manner. Further, NO production induced by anti-CR3 mAb was also inhibited by zymocel, beta-glucan with a high affinity to CR3. These results suggest that the CR3 molecule acts in a CD14-independent signaling pathway, and contributes to NO production by macrophages stimulated with high doses of LPS.
Matsuno
R
R
School of Pharmacy, Tokyo University of Pharmacy and Life Science, Japan.
Aramaki
Y
Y
Arima
H
H
Adachi
Y
Y
Ohno
N
N
Yadomae
T
T
Tsuchiya
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001666:Binding Sites, Antibody / Q000187:drug effects; D001667:Binding, Competitive / Q000276:immunology; D015153:Blotting, Western; D002417:Cattle; D004306:Dose-Response Relationship, Immunologic; D005312:Fetal Blood / Q000502:physiology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation / Q000187:drug effects; D016177:Macrophage-1 Antigen / Q000276:immunology / Q000502:physiology*; D008264:Macrophages / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis
D000911:Antibodies, Monoclonal; D008070:Lipopolysaccharides; D016177:Macrophage-1 Antigen; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase