Characterization of Zaire ebolavirus glycoprotein-specific monoclonal antibodies

Clin Immunol. 2011 Nov;141(2):218-27. doi: 10.1016/j.clim.2011.08.008. Epub 2011 Aug 31.

Abstract

Zaire ebolavirus (ZEBOV) can be transmitted by human-to-human contact and causes acute haemorrhagic fever with case fatality rates up to 90%. There are no effective therapeutic or prophylactic treatments available. The sole transmembrane glycoprotein (GP) is the key target for developing neutralizing antibodies. In this study, recombinant VSVΔG/ZEBOVGP was used to generate monoclonal antibodies (MAbs) against the ZEBOV GP. A total of 8 MAbs were produced using traditional hybridoma cell fusion technology, and then characterized by ELISA using ZEBOV VLPs, Western blotting, an immunofluorescence assay, and immunoprecipitation. All 8 MAbs worked in IFA and IP, suggesting that they are all conformational MAbs, however six of them recognized linearized epitopes by WB. ELISA results demonstrated that one MAb bound to a secreted GP (sGP 1-295aa); three bind to a part of the mucin domain (333-458aa); three MAbs recognized epitopes on the C-terminal domain of GP1 (296-501aa); and one bound to full length GP (VLPs/GP1,2 ΔTm). Using a mouse model these MAbs were evaluated for their therapeutic capacity during a lethal infection. All 8 MAb improved survival rates by 33%-100% against a high dose lethal challenge with mouse-adapted ZEBOV. This work has important implications for further development of vaccines and immunotherapies for ZEBOV infection.

MeSH terms

  • Animals
  • Antibodies, Monoclonal, Murine-Derived / biosynthesis
  • Antibodies, Monoclonal, Murine-Derived / immunology*
  • Antibodies, Viral / biosynthesis
  • Antibodies, Viral / immunology*
  • Antibody Specificity
  • Antigens, Viral / immunology*
  • Blotting, Western
  • Dose-Response Relationship, Immunologic
  • Ebolavirus / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / immunology
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Hemorrhagic Fever, Ebola / therapy
  • Humans
  • Hybridomas / immunology
  • Immunization, Passive
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / immunology*
  • Immunoprecipitation
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Protein Structure, Tertiary
  • Viral Envelope Proteins / chemistry
  • Viral Envelope Proteins / immunology*

Substances

  • Antibodies, Monoclonal, Murine-Derived
  • Antibodies, Viral
  • Antigens, Viral
  • Epitopes
  • Immunoglobulin G
  • Viral Envelope Proteins
  • envelope glycoprotein, Ebola virus