Identification of protective epitopes on ebola virus glycoprotein at the single amino acid level by using recombinant vesicular stomatitis viruses.
Ebola virus causes lethal hemorrhagic fever in humans, but currently there are no effective vaccines or antiviral compounds for this infectious disease. Passive transfer of monoclonal antibodies (MAbs) protects mice from lethal Ebola virus infection (J. A. Wilson, M. Hevey, R. Bakken, S. Guest, M. Bray, A. L. Schmaljohn, and M. K. Hart, Science 287:1664-1666, 2000). However, the epitopes responsible for neutralization have been only partially characterized because some of the MAbs do not recognize the short synthetic peptides used for epitope mapping. To identify the amino acids recognized by neutralizing and protective antibodies, we generated a recombinant vesicular stomatitis virus (VSV) containing the Ebola virus glycoprotein-encoding gene instead of the VSV G protein-encoding gene and used it to select escape variants by growing it in the presence of a MAb (133/3.16 or 226/8.1) that neutralizes the infectivity of the virus. All three variants selected by MAb 133/3.16 contained a single amino acid substitution at amino acid position 549 in the GP2 subunit. By contrast, MAb 226/8.1 selected three different variants containing substitutions at positions 134, 194, and 199 in the GP1 subunit, suggesting that this antibody recognized a conformational epitope. Passive transfer of each of these MAbs completely protected mice from a lethal Ebola virus infection. These data indicate that neutralizing antibody cocktails for passive prophylaxis and therapy of Ebola hemorrhagic fever can reduce the possibility of the emergence of antigenic variants in infected individuals.
Takada
Ayato
A
Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Japan.
Feldmann
Heinz
H
Stroeher
Ute
U
Bray
Mike
M
Watanabe
Shinji
S
Ito
Hiroshi
H
McGregor
Martha
M
Kawaoka
Yoshihiro
Y
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000276:immunology; D002460:Cell Line; D000939:Epitopes / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007116:Immunization, Passive; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011995:Recombination, Genetic; D017386:Sequence Homology, Amino Acid; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000276:immunology; D014764:Viral Proteins
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000939:Epitopes; D006023:Glycoproteins; D014764:Viral Proteins; C110613:secreted glycoprotein, Ebola virus