Characterization of contrasting features between hepatitis B virus genotype A and genotype D in small envelope protein expression and surface antigen secretion

Hepatitis B virus (HBV) genotypes A and D are prevalent in many parts of the world and show overlapping geographic distributions. We amplified the entire HBV genome from sera of patients with genotypes A and D and generated overlength constructs for transient transfection into Huh7 or HepG2 cells. Genotype D clones were associated with less HBsAg in culture supernatant and even less intracellular HBsAg. They produced less 2.1-kb RNA due to a weaker SPII promoter. Chimeric promoter constructs identified three divergent positions as most critical, and their exchange reversed extracellular HBsAg phenotype. The S protein of genotype D was more efficient at secretion, while its L protein possessed greater inhibitory effect. Swapping the S gene diminished genotypic difference in intracellular S protein but widened the difference in secreted HBsAg. In conclusion, HBV genotypes A and D differ in S protein expression, secretion and modulation by L protein.