This paper describes the biophysical and biochemical properties as well as electron microscopical studies of a candidate hepatitis A vaccine strain propagated in human fibroblast cells. Our results indicated that, in CsCl, the density of hepatitis A virus (HAV) from cell culture supernatant and of HAV extracted from infected cells was influenced by the quantity of lipid material associated with HAV. Antigenicity of untreated HAV, therefore, was detected primarily in low density CsCl fractions (1.11 g/ml, 1.21 g/ml). After lipid reduction with NP40 detergent or chloroform/Genetron, antigenicity and infectivity were primarily detected in high density CsCl fractions (1.31 g/ml). Electron microscopy demonstrated a strong association between membranous material and virus particles of low density in CsCl as well as virus-like particles in ultrathin sections of HAV-infected human fibroblast cells. The uncovered virus particles banded in the 1.31 g/ml dense CsCl fraction lacked lipid material. The value was 79 for 1.19 g/ml to 1.22 g/ml dense HAV and 147 for 1.29 g/ml to 1.33 g/ml HAV. Autoradiography of the radioiodinated dense HAV revealed some proteins with high (120K to 67K) and others with low (37K to 15K).


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