Complement-mediated neurotoxicity is regulated by homologous restriction.
The ability of beta-amyloid peptides to activate the classical complement cascade and the presence of various complement proteins including the membrane attack complex (C5b-9) on dystrophic neurites in Alzheimer's disease brains, raises the possibility that the complement system may contribute to this neurodegenerative disorder. To address this issue, we have studied the effect of complement activation on nerve growth factor (NGF)-differentiated rat pheochromocytoma PC12 cells, and on retinoic acid (RA)-differentiated human neuroblastoma SH-SY5Y cells. Although incubation of both cell types with human serum resulted in activation of complement, as indicated by iC3b formation, only PC12 but not SH-SY5Y cells were killed by human serum treatment. In contrast, heat-inactivated serum (56 degrees C, 45 min) was not neurotoxic. On SH-SY5Y cells, both PCR amplification and immunocytochemistry demonstrated the presence of CD59, a glycosylphosphatidylinositol-anchored protein that restricts homologous complement activation by inhibiting the formation of the membrane attack complex. The presence of CD59 probably accounts for the inability of human complement to lyse the human cell lines. Indeed, removal of glycosylphosphatidylinositol (GPI)-anchored proteins with phosphatidylinositol-specific phospholipase C (PI-PLC) rendered SH-SY5Y cells vulnerable to complement attack and eventually led to serum-medicated cell death. Reconstituted C5b-9 was also toxic to both PC12 and PI-PLC-pretreated SH-SY5Y cells. These observations suggest that complement activation can cause neuronal cell death and that this process is regulated by homologous restriction.
Shen
Y
Y
Neuroscience Department, Abbott Laboratories, Abbott Park, IL 60064-3500, USA.
Halperin
J A
JA
Lee
C M
CM
D000818:Animals; D001483:Base Sequence; D002470:Cell Survival / Q000187:drug effects; D003167:Complement Activation; D003169:Complement Inactivator Proteins / Q000494:pharmacology; D003165:Complement System Proteins / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008969:Molecular Sequence Data; D009474:Neurons / Q000201:enzymology / Q000502:physiology*; D016716:PC12 Cells; D010716:Phosphatidylinositols / Q000378:metabolism; D016133:Polymerase Chain Reaction; D051381:Rats; D013045:Species Specificity; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000378:metabolism
D003169:Complement Inactivator Proteins; D010716:Phosphatidylinositols; D003165:Complement System Proteins; D007770:L-Lactate Dehydrogenase; D010738:Type C Phospholipases