Functional identification of agretopic and epitopic residues within an HBcAg T cell determinant

Residues 120-131 within the hepatitis B core Ag (HBcAg) represent a dominant T cell recognition site for mice of the H-2S haplotype. This study was undertaken in order to identify residues within the p120-131 sequence which either interact with the TCR termed epitopic residues or interact with MHC class II molecules termed agretopic residues. For this purpose a panel of analogs of p120-131 composed of peptides containing single alanine substitutions for each residue was synthesized. These peptides were analyzed functionally for their ability to stimulate p120-131 or HBcAg-primed T cells and for their immunogenicity in B10.S or [B10.S X B10 (nonresponder)]F1 mice. Furthermore, analogs of p120-131 were used as stimulators and inhibitors of T cell activation in competitive inhibition experiments. Cumulatively these functional studies allowed us to identify residue 125 as a dominant epitopic residue and residues 127 and 129 as dominant agretopic residues. Furthermore, a p120-131 analog containing an alanine substitution for the dominant agretopic residue was immunogenic in B10.S mice, but was nonimmunogenic in (B10.S X B10)F1 mice indicating that T cell responsiveness is influenced by MHC class II gene dosage effects and can be inherited in an apparent recessive manner. In this study, critical residues involved in the immunogenicity of this dominant T cell determinant of HBcAg were defined, in a companion study, the influence of these residues on tolerogenicity was examined.