title: Ligation of human CD46 with purified complement C3b or F(ab')(2) of monoclonal antibodies enhances isoform-specific interferon gamma-dependent nitric oxide production in macrophages.
abstract: CD46, a complement regulatory protein widely expressed on human cells, serves as an entry receptor for measles virus (MV). We have previously shown that the expression of human CD46 in mouse macrophages restricts MV replication in these cells and enhances the production of nitric oxide (NO) in the presence of gamma interferon (IFN-gamma). In this study, we show that crosslinking human CD46 expressed on the mouse macrophage-like cell line RAW264.7 with purified C3b multimer but not monomer enhances NO production. The enhanced production of NO in response to IFN-gamma was observed again with C3b multimer but not monomer. The augmentation of NO production is human CD46-dependent with a CYT1>CYT2 profile. Thus, the reported MV-mediated NO production, irrespective of whether it is IFN-gamma-dependent or -independent, should be largely attributable to CD46 signaling but not to MV replication. Similar CYT1-dependent augmentation of NO production was reproducible with two CD46 ligating reagents, CD46-specific monoclonal antibodies (mAb) or their F(ab')(2) and MV hemagglutinin (H) and fusion (F) glycoproteins. Co-cultivation of mouse macrophages bearing human CD46 with Chinese hamster ovary (CHO) cells expressing MV H and F enhanced IFN-gamma-induced NO production. Yet, the NO levels induced by F(ab')(2) against CD46 or MV H/F on CHO cells were much lower than those induced by CD46-crosslinking mAb with Fc or MV infection. Removing the cytoplasmic tails of CD46 abrogated the augmentation of NO production triggered by all three stimulators. Thus, the CD46 CYT1 and CYT2 isoforms functionally diverge to elicit innate immune responses, which can be modulated by purified C3b multimer or anti-CD46 mAbs.
authors.affiliation: Department of Microbiology, University of Washington School of Medicine, Seattle, Washington, WA 98195, USA.
authors.forename: Akiko
authors.initials: A
authors.lastname: Hirano
authors.forename: Mitsue
authors.initials: M
authors.lastname: Kurita-Taniguchi
authors.forename: Yuko
authors.initials: Y
authors.lastname: Katayama
authors.forename: Misako
authors.initials: M
authors.lastname: Matsumoto
authors.forename: Timocy C
authors.initials: TC
authors.lastname: Wong
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Seya
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism*; D016466:CHO Cells / Q000378:metabolism; D002460:Cell Line; D003179:Complement C3b / Q000276:immunology / Q000378:metabolism*; D006224:Cricetinae; D006389:Hemagglutinins, Viral / Q000096:biosynthesis; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000378:metabolism*; D007371:Interferon-gamma / Q000494:pharmacology*; D008264:Macrophages / Q000276:immunology / Q000378:metabolism*; D008459:Measles virus / Q000378:metabolism; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis*; D020033:Protein Isoforms; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; C497857:CD46 protein, human; D006389:Hemagglutinins, Viral; D007140:Immunoglobulin Fab Fragments; C497859:Mcp protein, mouse; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D020033:Protein Isoforms; D009569:Nitric Oxide; D003179:Complement C3b; D007371:Interferon-gamma