Demonstration of Hepatitis B e Antigen in Hepatitis B Core Particles Obtained from the Nucleus of Hepatocytes Infected with Hepatitis B Virus

Authors:
  • HYH. Yoshizawa
    H. Yoshizawa
  • YIY. Itoh
    Y. Itoh
  • JSJ. P. Simonetti
    J. P. Simonetti
  • TTT. Takahashi
    T. Takahashi
  • AMA. Machida
    A. Machida
  • YMY. Miyakawa
    Y. Miyakawa
  • MMM. Mayumi
    M. Mayumi
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Abstract
Liver tissue infected with hepatitis B virus was homogenized and nuclei were separated by centrifugation. Hepatitis B core particles were obtained from the nucleus by the digestion with pronase followed by ultracentrifugation in a sucrose density gradient. Hepatitis B core particles were then treated with sodium dodecyl sulphate and 2 mercaptoethanol and tested for hepatitis B e antigen (HBeAg) by the haemagglutination method. The antigenicity of HBeAg was clearly demonstrated in hepatitis B core particles so treated, although untreated core particles did not reveal any detectable HBeAg activity. The localization of HBeAg in hepatitis B core particles was further supported by the results of a fluorescent antibody technique. When a frozen section of the liver infected with hepatitis B virus was stained with the specific rabbit antibody against HBeAg labelled with fluorescent isothiocyanate, only nuclei of hepatocytes were stained, in a similar distribution to hepatitis B core antigen visualized by fluorescent antibody against hepatitis B core antigen in a nearby section.