title: Selective and efficient inhibition of the alternative pathway of complement by a mAb that recognizes C3b/iC3b.
abstract: The alternative pathway (AP) of the complement system plays an important role in tissue damage and inflammation associated with certain autoimmune diseases and with ischemia-reperfusion injury. Selective inhibition of the AP could prevent such pathologies while allowing the classical and lectin pathways of complement activation to continue to provide protection. Here we present data describing selective inhibition of the AP of complement by anti-C3b/iC3b monoclonal antibody (mAb) 3E7, and by a chimeric, "deimmunized" form of this mAb, H17, which contains the human IgG1 Fc region and was further modified by substitution of amino acids in order to remove T cell epitopes. Both mAbs block AP-mediated deposition of C3b onto zymosan or Sepharose 4B, and they also inhibit AP-promoted lysis of rabbit erythrocytes. MAbs 3E7 and H17 also successfully compete with both factors B and H for binding to C3b-opsonized substrates, and the ability of both mAbs to inhibit the AP is blocked by pre-incubation with two different sources of C3(H2O). Kinetic measurements demonstrate that mAb 3E7 effectively stops progression of C3b deposition after AP activation is initiated. Our results therefore suggest that these mAbs block activation of the AP by binding to both C3(H2O) and to C3b, and thus prevent binding and activation of factor B. Based on these and other observations, mAb H17 may find future use in therapeutic applications focused on selective inhibition of the AP.
authors.affiliation: Department of Biochemistry and Molecular Genetics, Box 800733, University of Virginia Health Sciences Center, School of Medicine, Charlottesville, VA 22908-0733, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: DiLillo
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Pawluczkowycz
authors.forename: Wu
authors.initials: W
authors.lastname: Peng
authors.forename: Adam D
authors.initials: AD
authors.lastname: Kennedy
authors.forename: Paul V
authors.initials: PV
authors.lastname: Beum
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Lindorfer
authors.forename: Ronald P
authors.initials: RP
authors.lastname: Taylor
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology; D003179:Complement C3b / Q000037:antagonists & inhibitors*; D011415:Complement Factor B / Q000037:antagonists & inhibitors; D017242:Complement Factor H / Q000037:antagonists & inhibitors; D003170:Complement Pathway, Alternative / Q000187:drug effects*; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D003179:Complement C3b; D017242:Complement Factor H; D011415:Complement Factor B