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Medical Molecular Morphology

, Volume 39, Issue 2, pp 63–71 | Cite as

Morphology of hepatitis C and hepatitis B virus particles as detected by immunogold electron microscopy

  • Masahiko Kaito
  • Satoshi Ishida
  • Hideaki Tanaka
  • Shinichiro Horiike
  • Naoki Fujita
  • Yukihiko Adachi
  • Michinori Kohara
  • Masayoshi Konishi
  • Shozo Watanabe
SPECIAL REVIEW SERIES: Molecular morphological approach for pathogenesis of hepatogastrointestinal diseases

Abstract

We performed indirect immunogold electron microscopy (EM) for immunological identification and characterization of hepatitis C virus (HCV). To clarify the morphology of HCV, an indirect immunogold EM of two plasma samples from patients with high HCV RNA titers was carried out using antibodies specific for the putative HCV envelope protein (E) 1. Spherical virus particles 55–65 nm in diameter with delicate spike projections were detected in the 1.14–1.16 g/ml fractions after sucrose density gradient centrifugation. Polyclonal and monoclonal antibodies to the putative HCV E1 specifically recognized these particles. In addition, immunogold EM of the samples was also performed to uncover the morphology of HCV core particles. Spherical particles 33–40 nm in diameter (average, 37 nm) were detected in the 1.22- to 1.25-g/ml fractions by conventional EM after sucrose density gradient centrifugation. Immunogold EM using rabbit polyclonal antibody (RR8) specific for the putative HCV core protein and colloidal gold-labeled goat antirabbit IgG showed binding of the gold particles with RR8. Some of the HCV core particles showed icosahedric morphology. Optical rotation technique showed that the HCV core particles exhibit sixfold symmetry and that the length of the regular hexagon side is approximately 20 nm, suggesting that they have an icosahedric structure. Further, the detection limit of the indirect immunogold EM was evaluated in 11 plasma samples from chronic hepatitis B patients with different degrees of hepatitis B virus (HBV) DNA titers using antihepatitis B surface antigen antibody. The study showed that the detection limit of virus using this method is 107 virions/ml.

Key words

HBV HCV Immune electron microscopy Colloidal gold Buoyant density Optical rotation technique 

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Copyright information

© The Japanese Society for Clinical Molecular Morphology 2006

Authors and Affiliations

  • Masahiko Kaito
    • 1
  • Satoshi Ishida
    • 1
  • Hideaki Tanaka
    • 1
  • Shinichiro Horiike
    • 1
  • Naoki Fujita
    • 1
  • Yukihiko Adachi
    • 1
  • Michinori Kohara
    • 2
  • Masayoshi Konishi
    • 3
  • Shozo Watanabe
    • 3
  1. 1.Department of Gastroenterology and Hepatology, Division of Clinical Medicine and Biomedical Science, Institute of Medical ScienceMie University Graduate School of MedicineMieJapan
  2. 2.Department of Microbiology and Cell BiologyThe Tokyo Metropolitan Institute of Medical ScienceTokyoJapan
  3. 3.Health Administration CenterMie UniversityMieJapan

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