a therapeutic antibody against west nile virus neutralizes infection by blocking fusion within endosomes治疗感染西尼罗河病毒中和抗体通过阻断在核内体融合.pdfVIP

A Therapeutic Antibody against West Nile Virus Neutralizes Infection by Blocking Fusion within Endosomes 1 2 2 2 1,3,4 Bruce S. Thompson , Bastiaan Moesker , Jolanda M. Smit , Jan Wilschut , Michael S. Diamond *, Daved H. Fremont1* 1 Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, United States of America, 2 Department of Medical Microbiology-Molecular Virology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands, 3 Medicine, Washington University School of Medicine, St. Louis, Missouri, United States of America, 4 Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, United States of America Abstract Defining the precise cellular mechanisms of neutralization by potently inhibitory antibodies is important for understanding how the immune system successfully limits viral infections. We recently described a potently inhibitory monoclonal antibody (MAb E16) against the envelope (E) protein of West Nile virus (WNV) that neutralizes infection even after virus has spread to the central nervous system. Herein, we define its mechanism of inhibition. E16 blocks infection primarily at a post- attachment step as antibody-opsonized WNV enters permissive cells but cannot escape from endocytic compartments. These cellular experiments suggest that E16 blocks the acid-catalyzed fusion step that is required for nucleocapsid entry into the cytoplasm. Indeed, E16 directly inhibits fusion of WNV with liposomes. Additionally, low-pH exposure of E16–WNV complexes in the absence of target membranes did not fully inactivate infectious virus, further suggesting that