Morphological identification of hepatitis C virus E1 and E2 envelope glycoproteins on the virion surface using immunogold electron microscopy.
It is known that hepatitis C virus (HCV) particles are spherical, 55-65 nm particles with fine surface projections of about 6 nm in length and with a 30-35 nm inner core. We have reported that free HCV particles labeled with gold particles specific to the HCV E1 glycoprotein are located in 1.14-1.16 g/ml fractions from plasma samples with high HCV RNA titers after sucrose density gradient centrifugation. However, the morphology of the HCV E2 glycoprotein on the virion has not yet been elucidated. To visualize HCV E2 localization on the virion, we used the same plasma samples where HCV particles were clearly shown. An indirect immunogold electron microscopic study was carried out using monoclonal and polyclonal anti-HCV E2 antibodies. HCV-like particles specifically reacted with the anti-HCV E2 antibodies. Moreover, to evaluate the localization of the HCV E1 and E2 glycoproteins on the virion surface, an immunogold electron microscopic study using double labeling with anti-HCV E1 antibodies and anti-HCV E2 antibodies was also performed. These particles also specifically reacted with both anti-E1 and E2 antibodies. This is the first report showing the presence of both HCV E1 and E2 glycoproteins on HCV virion surface in human plasma samples.
Kaito
Masahiko
M
Department of Gastroenterology and Hepatology, Division of Clinical Medicine and Biomedical Science, Institute of Medical Science, Mie University Graduate School of Medicine, Mie 514-8507, Japan. kaitoma@clin.medic.mie-u.ac.jp
Watanabe
Shozo
S
Tanaka
Hideaki
H
Fujita
Naoki
N
Konishi
Masayoshi
M
Iwasa
Motoh
M
Kobayashi
Yoshinao
Y
Gabazza
Esteban Cesar
EC
Adachi
Yukihiko
Y
Tsukiyama-Kohara
Kyoko
K
Kohara
Michinori
M
D016174:Hepacivirus / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron / Q000379:methods; D012367:RNA, Viral / Q000097:blood; D014759:Viral Envelope Proteins / Q000032:analysis / Q000648:ultrastructure*; D014771:Virion / Q000737:chemistry / Q000648:ultrastructure*
C087035:E1 protein, Hepatitis C virus; D012367:RNA, Viral; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus