A single amino acid substitution in the S1 and S2 Spike protein domains determines the neutralization escape phenotype of SARS-CoV.
In response to SARS-CoV infection, neutralizing antibodies are generated against the Spike (S) protein. Determination of the active regions that allow viral escape from neutralization would enable the use of these antibodies for future passive immunotherapy. We immunized mice with UV-inactivated SARS-CoV to generate three anti-S monoclonal antibodies, and established several neutralization escape mutants with S protein. We identified several amino acid substitutions, including Y442F and V601G in the S1 domain and D757N and A834V in the S2 region. In the presence of each neutralizing antibody, double mutants with substitutions in both domains exhibited a greater growth advantage than those with only one substitution. Importantly, combining two monoclonal antibodies that target different epitopes effected almost complete suppression of wild type virus replication. Thus, for effective passive immunotherapy, it is important to use neutralizing antibodies that recognize both the S1 and S2 regions.
Mitsuki
Yu-ya
YY
Department of Immunology, National Institute of Infectious Diseases, Toyama 1-23-1, Tokyo 162-8640, Japan.
Ohnishi
Kazuo
K
Takagi
Hirotaka
H
Oshima
Masamichi
M
Yamamoto
Takuya
T
Mizukoshi
Fuminori
F
Terahara
Kazutaka
K
Kobayashi
Kazuo
K
Yamamoto
Naoki
N
Yamaoka
Shoji
S
Tsunetsugu-Yokota
Yasuko
Y
D019943:Amino Acid Substitution / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology*; D051379:Mice; D020125:Mutation, Missense; D009500:Neutralization Tests; D017434:Protein Structure, Tertiary; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000254:growth & development / Q000276:immunology*; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology*; D010948:Viral Plaque Assay; D014779:Virus Replication / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D008562:Membrane Glycoproteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins; C578557:spike glycoprotein, SARS-CoV; C578553:spike protein, mouse hepatitis virus