title: Prevention of experimental autoimmune myasthenia gravis by rat Crry-Ig: A model agent for long-term complement inhibition in vivo.
abstract: Despite its vital role in innate immunity, complement is involved in a number of inflammatory pathologies and has therefore become a therapeutic target. Most agents generated for anti-complement therapy have short half-lives in plasma, or have been of mouse or human origin, thereby limiting their use either to murine models of disease or to short-term therapy. Here we describe the generation of a long-acting rat therapeutic agent based on the rat complement inhibitor, Crry. Characterisation of various soluble forms of Crry demonstrated that the amino-terminal four short-consensus repeat domains were required for full regulatory and C3b-binding activities. Fusion of these domains to rat IgG2a Fc generated an effective complement inhibitor (rCrry-Ig) with a circulating half-life prolonged from 7 min for Crry alone to 53 h for rCrry-Ig. Systemic administration of rCrry-Ig over 5 weeks generated a weak immune response to the recombinant agent, however this was predominantly IgM in nature and did not neutralise Crry function or cause clearance of the agent from plasma. Administration of rCrry-Ig completely abrogated clinical disease in a rat model of myasthenia gravis whereas soluble Crry lacking the immunoglobulin Fc domain caused a partial response. rCrry-Ig not only ablated clinical disease, but also prevented C3 and C9 deposition at the neuromuscular junction and inhibited cellular infiltration at this site. The long half-life and low immunogenicity of this agent will be useful for therapy in chronic models of inflammatory disease in the rat.
authors.affiliation: Department of Medical Biochemistry and Immunology, School of Medicine, Cardiff University, Heath Park, Cardiff CF14 4XN, United Kingdom.
authors.forename: Natalie J
authors.initials: NJ
authors.lastname: Hepburn
authors.forename: Jayne L
authors.initials: JL
authors.lastname: Chamberlain-Banoub
authors.forename: Anwen S
authors.initials: AS
authors.lastname: Williams
authors.forename: B Paul
authors.initials: BP
authors.lastname: Morgan
authors.forename: Claire L
authors.initials: CL
authors.lastname: Harris
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000097:blood / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*; D003179:Complement C3b / Q000276:immunology; D003169:Complement Inactivator Proteins / Q000494:pharmacology*; D003165:Complement System Proteins / Q000276:immunology*; D005260:Female; D006207:Half-Life; D007074:Immunoglobulin G / Q000494:pharmacology*; D018482:Muscle, Skeletal / Q000187:drug effects / Q000473:pathology; D020720:Myasthenia Gravis, Autoimmune, Experimental / Q000517:prevention & control*; D009469:Neuromuscular Junction / Q000187:drug effects / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D011956:Receptors, Cell Surface / Q000097:blood / Q000276:immunology / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000097:blood / Q000302:isolation & purification / Q000494:pharmacology; D012995:Solubility / Q000187:drug effects; D020349:Surface Plasmon Resonance
chemical_list: D000954:Antigens, Surface; D003169:Complement Inactivator Proteins; C513105:Cr1l protein, rat; D007074:Immunoglobulin G; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D003179:Complement C3b; D003165:Complement System Proteins