Experimental infection of rats with Borna disease virus (BDV) and natural BDV infection of horses and sheep leads to a virus-induced T-cell-mediated immunopathology in the central nervous system. Earlier work revealed the importance of the BDV-specific T-cell response and of CD8(+) effector cells in particular in the destruction of virus-infected cells. Evidence was also presented that this major histocompatibility complex class I-restricted lysis detected in vitro might play a functional role in the immunopathogenesis of Borna disease. The present study employed different vaccinia virus recombinants expressing single BDV-specific proteins to investigate the specificity of the cytolytic CD8(+)-T-cell response, revealing a major epitope on the BDV nucleoprotein p40. In contrast, no direct evidence in favor of the presence of in vivo relevant cytotoxic T-lymphocyte epitopes on other BDV-specific proteins was found.