Direct evidence that naturally occurring mutations within hepatitis B core epitope alter CD4+ T-cell reactivity

Exacerbations of chronic hepatitis B have been associated with accumulation of mutations in the HBV core gene, with amino acid (aa) substitutions clustering between aa 50 and 69. This region of the nucleocapsid protein is known as an immunodominant epitope for CD4+ T-lymphocytes, however the impact of these mutations on T-cell reactivity has not been investigated. For this purpose, we undertook fine mapping of the reactivity of peripheral blood lymphocytes, isolated from patients with acute (n = 8) or chronic hepatitis B (n = 10), against a panel of branched synthetic peptides. The peptide aa sequences corresponded to the wild type HBV (aa 50-69), or contained 1-3 aa changes derived on the basis of naturally occurring mutations. In four of eight patients with acute hepatitis B the wild type peptide 50-69, which corresponded to the core gene sequence of HBV present in these patients, induced a strong T-cell proliferative response. In the same cases, the T-cell response to the mutant peptides was altered at various degrees, depending on the number and the position of aa changes. The most pronounced inhibition of CD4+ T-cell response (between 44 and 92%) was caused by a peptide ligand with two aa substitutions at positions 64 and 67. These results demonstrate that mutations within immunodominant epitopes of the HBV nucleocapsid can affect the CD4+ T-lymphocyte reactivity, which may have a role for the accumulation of certain HBV strains after hepatitis flares during the course of chronic HBV infection.