The ultrastructural morphology of native hepatitis B virus |
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Authors: | Masahiko Kaito Hiroyoshi Ohba Joe Chiba Michinori Kohara Hideaki Tanaka Naoki Fujita Esteban Cesar Gabazza Shozo Watanabe Masayoshi Konishi Yukihiko Adachi |
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Institution: | (1) Department of Gastroenterology and Hepatology, Division of Clinical Medicine and Biomedical Science, Institute of Medical Science, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu, Mie 514-8507, Japan;(2) Department of Biological Science and Technology, Tokyo University of Science, Chiba, Japan;(3) Department of Microbiology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan;(4) Health Administration Center, Mie University, Mie, Japan |
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Abstract: | Cell lines (2.2.15 cells) capable of supporting the replication of hepatitis B virus (HBV) DNA and intact viral particles
have been established by HBV DNA transfection into HepG2 cells. The purpose of this study was to determine the ultrastructural
morphology of native HBV particles without purification in the culture supernatants and in sera from patients. Electron microscopy
(EM) and immunogold EM of the samples were carried out using polyclonal and monoclonal anti-hepatitis B surface antigen antibodies.
HBV particles in the purified samples from the culture supernatants by density-gradient centrifugation were examined to compare
the morphology with that of unpurified samples. EM and immunogold EM studies demonstrated the presence of Dane particles (41.8 nm
in diameter), cobra-shaped (head diameter, 42.4 nm), and horn-shaped (head diameter, 43.5 nm) particles in the culture supernatants
and in the sera from two patients. The tail of the cobra-like particles had a diameter of 21.0 nm and a length of 214 nm.
The hornlike particles had a long branch 20.1 nm in diameter with a length of 189 nm, and a short branch 21.4 nm in diameter
with a length of 112 nm. The ratio of Dane particles and cobra- and horn-shaped particles in the supernatants was 5 : 4 : 1.
After ultracentrifugation, the cobra- and horn-shaped particles completely disappeared; there were only Dane particles together
with spheres of 22 nm and filaments. In conclusion, this study showed for the first time that the native replicative form
of HBV is cobra- and horn-shaped. |
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Keywords: | Hepatitis B virus Particle Electron microscopy Immunoelectron microscopy Cell culture 2 2 15 cells Human serum |
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