Demonstration of hepatitis B e antigen in hepatitis B core particles obtained from the nucleus of hepatocytes infected with hepatitis B virus.
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.
D002467:Cell Nucleus / Q000276:immunology / Q000382:microbiology*; D005455:Fluorescent Antibody Technique; D006386:Hemagglutination Tests; D006509:Hepatitis B / Q000382:microbiology*; D006511:Hepatitis B Antigens / Q000032:analysis* / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000276:immunology*; D006801:Humans; D008099:Liver / Q000166:cytology*
D006511:Hepatitis B Antigens