Macrophage phagocytosis of myelin in vitro determined by flow cytometry: phagocytosis is mediated by CR3 and induces production of tumor necrosis factor-alpha and nitric oxide.
Demyelination of axons in the central nervous system (CNS) during multiple sclerosis (MS) and its animal model experimental allergic encephalomyelitis (EAE) is a result of phagocytosis and digestion by macrophages (M phi) and the local release of inflammatory mediators like tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO). We have investigated the process of myelin phagocytosis by M phi in vitro using flow cytometric analysis. The binding and uptake of CNS-derived myelin was dose dependent, was abolished in the presence of EDTA and was enhanced after opsonization with complement. The phagocytosis of opsonized myelin could be inhibited by antibodies directed against complement receptor type 3 (CR3). Furthermore, CR3 also contributes to phagocytosis of non-opsonized myelin, e.g. under serum-free conditions. The phagocytosis of CNS-derived myelin induced the production of substantial amounts of TNF-alpha and NO by the M phi. Our results indicate an important role for CR3 in myelin phagocytosis. The induction of TNF-alpha and NO which accompanies this phagocytosis may further contribute to the overall process of demyelination during MS or EAE.
Department of Immunology and Cell Biology, Faculty of Medicine, Vrije Universiteit, Amsterdam, The Netherlands. l.van-der-laan.cell@med.vu.nl
L J
LJ
van der Laan
S R
SR
Ruuls
K S
KS
Weber
I J
IJ
Lodder
E A
EA
Döpp
C D
CD
Dijkstra
D000818:Animals; D001921:Brain / Q000276:immunology; D003176:Complement C3 / Q000378:metabolism; D005434:Flow Cytometry; D007111:Immunity, Cellular; D016177:Macrophage-1 Antigen / Q000502:physiology*; D008264:Macrophages / Q000276:immunology*; D008854:Microscopy, Electron; D009186:Myelin Sheath / Q000276:immunology*; D009569:Nitric Oxide / Q000096:biosynthesis; D010587:Phagocytosis; D051381:Rats; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D003176:Complement C3; D016177:Macrophage-1 Antigen; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide