Selective expression of immunogenic, virus-like particle-derived antibody-binding epitopes

The incorporation of linear and conformational antibody-binding epitopes into polyepitope, chimeric antigens with satisfactory immunogenicity is a challenge. We selectively expressed antigen fragments encoding the (linear e2 epitope (C_79-149) of hepatitis B virus (pre)core antigen (HBc/eAg) and the conformational 'a' epitope (S_80-180) of hepatitis B surface antigen (HBsAg) in a novel) system. The domains were expressed as chimeric antigens containing either heat shock protein (hsp)73-binding simian virus 40 large tumor antigen (e.g. T₇₇) or non-hsp-binding (e.g. T₆₀) sequences at their N-termini. We compared their type of expression with their immunogenicity for B cells (when delivered as a DNA vaccine). The type of expression investigated included their level of expression, the secretion or intracellular expression of the antigen and the stress protein (hsp)-associated versus nonassociated expression. The linear e2 epitope of HBc/eAg was efficiently expressed as an intracellular, hsp73-binding fusion protein, and efficiently primed an HBc/eAg-specific antibody response when delivered in this form. The conformational 'a' epitope of HBsAg most efficiently stimulated B cells as a secreted, non-hsp-associated fusion protein. These data demonstrate that different B cell-stimulating epitopes of vaccine-relevant viral antigens can be selectively isolated and expressed in suitable expression systems, but that the requirements that have to be fulfilled to obtain optimal immunogenicity differ strikingly between individual epitopes.