Characterization of the nucleocapsid protein of Hantaan virus strain 76-118 using monoclonal antibodies.
We characterized the antigenic sites on the nucleocapsid protein (NP) of Hantaan virus (HTN) using 10 monoclonal antibodies (MAbs). At least seven antigenic sites were revealed by a competitive binding assay and divided into three partially overlapping antigenic regions (I, II and III). Regions I [amino acids (aa) 1-103], II (aa 104-204) and III (aa 205-402) were mapped on NP by examining the reactivity of truncated gene products. Those that corresponded to region I reacted with immune mouse serum, indicating that the region contained major linear epitopes as reported with Four corners virus (FCV) and Puumala virus (PUU) NP. At least one MAb to each region inhibited viral growth when they were introduced into cells by scrape-loading. In addition, they conferred protection from a lethal HTN challenge to newborn mice. A PEPSCAN assay localized the epitope of MAb E5/G6 between aa 166-175. Since E5/G6, which had the highest inhibitory effect both in cells and in mice, showed no virus neutralization activity by ordinary neutralization test, this region is suggested to be important for the virus growth after entry into the cells.
Yoshimatsu
K
K
Institute for Animal Experimentation, School of Medicine, Hokkaido University, Japan.
Arikawa
J
J
Tamura
M
M
Yoshida
R
R
Lundkvist
A
A
Niklasson
B
B
Kariwa
H
H
Azuma
I
I
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D001483:Base Sequence; D012831:Bombyx / Q000166:cytology; D002213:Capsid / Q000276:immunology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D017931:DNA Primers; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D005260:Female; D018063:Hantaan virus / Q000235:genetics / Q000276:immunology*; D006480:Hemorrhagic Fever with Renal Syndrome / Q000517:prevention & control; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011247:Pregnancy; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014709:Vero Cells; D014758:Viral Core Proteins / Q000276:immunology*; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D017931:DNA Primers; D000939:Epitopes; D011994:Recombinant Proteins; D014758:Viral Core Proteins