title: Restoration of complement-enhanced neutralization of vaccinia virus virions by novel monoclonal antibodies raised against the vaccinia virus complement control protein.
abstract: The vaccinia virus complement control protein (VCP) is secreted by infected cells and has been shown to inhibit complement activation through interactions with C3b/C4b. It contains four short consensus repeat (SCR) domains. It has been suggested that all four SCRs are required for VCP's activity. To elucidate which SCR domains are involved in abolishing complement-enhanced neutralization of vaccinia virus virions, we generated and characterized a panel of mouse monoclonal antibodies (MAbs) raised against VCP. Ten MAbs were isolated and all recognized VCP on Western blots under reducing conditions as well as native-bound VCP in a sandwich enzyme-linked immunosorbent assay. Three of the 10 MAbs (2E5, 3D1, and 3F11) inhibited VCP's abolition of complement-enhanced neutralization of vaccinia virus virions. These MAbs blocked the interaction of VCP with C3b/C4b. The seven remaining MAbs did not alter VCP function in the complement neutralization assay and recognized VCP bound to C3b/C4b. To understand MAb specificity and mode of interaction with VCP, we mapped the MAb binding regions on VCP. The seven nonblocking MAbs all bound to the first SCR of VCP. One of the blocking MAbs recognized SCR 2 while the other two recognized either SCR 4 or the junction between SCRs 3 and 4, indicating that structural elements involved in the interaction of VCP with C3b/C4b are located within SCR domains 2 and 3 and 4. These anti-VCP MAbs may have clinical significance as therapeutic inhibitors of VCP's complement control activity and may also offer a novel approach to managing vaccinia virus vaccine complications that occur from smallpox vaccination.
authors.affiliation: Department of Medicine, Division of Infectious Diseases, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Stuart N
authors.initials: SN
authors.lastname: Isaacs
authors.forename: Emelia
authors.initials: E
authors.lastname: Argyropoulos
authors.forename: Georgia
authors.initials: G
authors.lastname: Sfyroera
authors.forename: Shamim
authors.initials: S
authors.lastname: Mohammad
authors.forename: John D
authors.initials: JD
authors.lastname: Lambris
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D002522:Chlorocebus aethiops; D003179:Complement C3b / Q000276:immunology* / Q000378:metabolism; D015935:Complement C4b / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D014616:Vaccinia virus / Q000276:immunology*; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D014771:Virion / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; C066459:complement-control protein, Vaccinia virus; D003179:Complement C3b; D015935:Complement C4b