C5 inhibitor rEV576 protects against neural injury in an in vitro mouse model of Miller Fisher syndrome.
Guillain-Barré syndrome and its clinical variants, including the anti-GQ1b ganglioside-mediated Miller Fisher syndrome (MFS), comprise the world's leading cause of acute neuromuscular paralysis. Presently, no specific drug therapies exist. The complement cascade, which is activated in these patients, forms an attractive drug target. In this study, we tested whether the complement C5-inhibiting recombinant protein, rEV576, was able to prevent neural injury in a previously developed in vitro mouse model for MFS. Mouse hemidiaphragm preparations were treated with anti-GQ1b antibody and normal human serum as a source of complement with added rEV576 or control protein. Immunohistology in control tissue showed deposition of C3c and membrane attack complex at neuromuscular junctions (NMJs), along with terminal motor axonal neurofilament degradation as well as ethidium homodimer-2 staining showing perisynaptic Schwann cell (pSC) injury. Electrophysiological and functional analyses showed block of synaptic transmission at the NMJ after an initial period of a dramatically high level of asynchronous acetylcholine release. In tissue treated with rEV576, all these indicators of motor neuronal damage were absent, except for the presence of C3c, indicating effective inhibition of C5. These results demonstrate that rEV576 effectively prevents development of neuronal and pSC damage in experimental murine neuropathy.
Halstead
Susan K
SK
Divison of Clinical Neurosciences, Glasgow Biomedical Research Centre, University of Glasgow, Glasgow, UK.
Humphreys
Peter D
PD
Zitman
Femke M P
FM
Hamer
John
J
Plomp
Jaap J
JJ
Willison
Hugh J
HJ
D000818:Animals; D003182:Complement C5 / Q000037:antagonists & inhibitors*; D003169:Complement Inactivator Proteins / Q000494:pharmacology*; D015938:Complement Membrane Attack Complex / Q000187:drug effects*; D003964:Diaphragm / Q000187:drug effects; D004195:Disease Models, Animal; D005071:Evoked Potentials / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D019476:Insect Proteins / Q000494:pharmacology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D019846:Miller Fisher Syndrome / Q000188:drug therapy*; D009469:Neuromuscular Junction / Q000187:drug effects*; D009924:Organ Culture Techniques; D011994:Recombinant Proteins / Q000494:pharmacology
D003182:Complement C5; D003169:Complement Inactivator Proteins; D015938:Complement Membrane Attack Complex; D019476:Insect Proteins; D011994:Recombinant Proteins