Integrin Mac-1 and beta-amyloid in microglial release of nitric oxide.
The beta-amyloid protein associated with Alzheimer's disease (AD) has been well characterized biochemically; however, its primary biological function and mode of action in AD has not been determined. We have shown previously that beta-amyloid (beta25-35), in combination with interferon-gamma (IFN-gamma), can induce nitric oxide release from cultured hippocampal microglial cells. In the present study, binding of beta-amyloid with the leukocyte integrin Mac-1, a cell surface receptor on microglia, was studied by observing (1) inhibition of beta-amyloid (beta25-35)-mediated release of nitric oxide from cultured microglial cells following exposure to monoclonal antibodies against Mac-1 (anti-CD18 and anti-CD11b) and (2) competitive binding of fluorochrome-labeled beta25-35 with anti-CD18 or anti-CD11b using fluorescent flow cytometry. Wt.3 (anti-CD18 antibody) and OX42 (anti-CD11b antibody) were as effective as opsonized zymosan at inducing the release of nitric oxide from microglia. Furthermore, Wt.3 and OX42 acted synergistically to induce maximum nitric oxide release. An interaction between beta-amyloid and CD18 of Mac-1 was evidenced by the suppressive action of beta25-35 on Wt.3-mediated release of nitric oxide and the synergistic action between OX42 and beta25-35 in inducing nitric oxide release from microglia. The tissue culture study was supported by competitive binding assays of fluorochrome-labeled beta25-35 and Mac-1 antibodies (Wt.3 or OX42). The majority of microglial cells (71%) did bind biotinylated beta-amyloid in the presence of cytochalasin B, suggesting that beta-amyloid binding to microglia is a receptor-mediated event. Furthermore, pre-exposure to Wt.3, but not OX42, significantly decreased binding of biotinylated beta25-35 to microglia. These findings suggest that CD18 of Mac-1 may play a role in beta-amyloid-mediated release of nitric oxide.
Goodwin
J L
JL
Department of Anatomy, Iowa State University, Ames 50011, USA.
Kehrli
M E
ME
Uemura
E
E
D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D019921:Biotinylation; D018821:CD18 Antigens / Q000276:immunology; D002478:Cells, Cultured; D004790:Enzyme Induction; D016177:Macrophage-1 Antigen / Q000378:metabolism*; D017628:Microglia / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis*; D009573:Nitrites / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D016229:Amyloid beta-Peptides; D018821:CD18 Antigens; D016177:Macrophage-1 Antigen; D009573:Nitrites; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase