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Hepatitis delta virus (HDV) is a defective virus that requires the supply of hepatitis B virus surface antigen (HBsAg) for replication and transmission. We have previously demonstrated that co-transduction of BHK cells with Bac- GD, a recombinant baculovirus expressing large hepatitis delta antigen (L-HDAg), and Bac-GS2, another recombinant baculovirus expressing HBsAg, gives rise to the assembly and secretion of 22 nm HBsAg subviral particles and 35-37 nm HDV-like particles (HDV VLP). In this study we uncovered oversize particles (>50 nm in diameter) comprised of HBsAg and L-HDAg and the particle properties varied with the relative dosages of Bac-GD and Bac-GS2, as demonstrated by Western blot, transmission electron microscopy and immunogold labeling. At a given Bac-GS2 dosage, decreasing the Bac-GD dosage resulted in the expression of more HBsAg, elevated secretion of HBsAg subviral particles, incorporation of more HBsAg into the HDV VLP, narrower particle size distribution and lower particle density. These data collectively demonstrated that the composition, and hence the properties, of HDV VLPs could be manipulated by altering the relative expression levels of structure proteins.

Varied Properties of Hepatitis-Delta Virus-like Particles Produced by Baculovirus-Transduced Mammalian Cells