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Full and empty particles of hepatitis B virus in hepatocytes from patients with HBsAg-positive chronic active
Abstract:
Complete and defective hepatitis B virus (HBV) particles in sera and hepatocytes were observed by electron microscopy for an understanding of the maturation process of hepatitis B virus. To distinguish Dane particles with or without DNA on the basis of staining density with uranyl acetate, Dane particles, purified from sera of asymptomatic carriers, and Dane particle cores, separated by ultracentrifugation in a metrizamide density gradient, were observed by electron microscopy. Complete cores at a low density (1.19 to 1.23 gm/cm3) were electron dense and incomplete cores at a high density (1.23 to 1.27 gm/cm3) were partially electron dense or empty. These findings demonstrated that the presence or absence of DNA is reflected by the electron density of the core. Our result, in which less than 10% serum Dane particles have full cores in ultrathin sections of the pellet, is in agreement with Gerin's finding that defective Dane particles are predominent in sera. Naked core particles and Dane particles in two biopsy specimens from patients with hepatitis B surface antigen, hepatitis B e antigen, and DNA polymerase-positive, chronic active hepatitis were classified into complete versus incomplete particles on the basis of electron density. Nine hundred and fourteen naked core particles were detected in nuclei and cytosol of 68 hepatocytes. One hundred and five core particles (11.5%) were electron dense and 809 core particles (88.5%) were partially electron dense or empty. Furthermore, 488 Dane particles were observed in the cisternae of the endoplasmic reticulum of these hepatocytes. Fifty Dane particles (10.2%) had full cores, and 438 Dane particles (89.8%) had partially full or empty cores. These findings suggest that DNA may be incorporated into about 1 to 10% of core particles when they are assembled in nuclei of hepatocytes. Morphologic differences in damage to hepatocytes containing various frequencies of full Dane particles were also studied, but no significant correlation was found between damage in hepatocytes and frequency of full Dane particles.