A virulent Salmonella expressing hybrid hepatitis B virus core/pre-S genes for oral vaccination.

This report reviews and extends data on the use of hepatitis B virus (HBV) core (HBcAg) particles as a carrier moiety for B-cell epitopes of the HBV envelope proteins. Virus-neutralizing epitopes of the HBV pre-S region were inserted at the N-terminus, the N-terminus through a precore linker sequence, the C-terminus and an internal position of HBcAg by genetic engineering in Escherichia coli. The hybrid HBc/pre-S proteins were purified and their antigenicity and immunogenicity analysed. All purified HBc/pre-S particles were particulate. Pre-S epitopes inserted at the N-terminus through a precore polylinker, the truncated C-terminus and at the internal position between HBcAg amino acids 75 and 81 were accessible on the particle surface. N-terminal fusions required the presence of the linker sequence to become surface accessible and immunogenic. Fusions to the N- and C-termini of HBcAg did not interfere with HBcAg antigenicity and immunogenicity. In contrast, insertion at the internal site abrogated recognition of HBcAg by five out of six monoclonal antibodies and diminished recognition by human polyclonal anti-HBc antibodies as well as HBcAg immunogenicity. A pre-S(2) sequence fused to the C-terminus of HBcAg was surface accessible and weakly immunogenic. Pre-S(1) sequences fused to the N-terminus through a precore linker were surface accessible and highly immunogenic. The same sequence fused to the core methionine was not surface accessible or immunogenic. Insertion of the same pre-S(1) sequence at an internal position of HBcAg resulted in the most efficient anti-pre-S(1) antibody response.(ABSTRACT TRUNCATED AT 250 WORDS)