Transgenic (Tg) mice expressing HLA class I alleles and lacking murine MHC class I represent a useful model for the pre-clinical evaluation of human
vaccines, which focus on induction of CD8(+) T-cell responses. We have developed a platform to be used in Tg mice for exploring the immunogenicity of T-cell targets, whose immunologic
epitopes have yet to be defined. To test the attributes of the evaluation system in the context of an important human pathogen, we have explored multiple
antigens from cytomegalovirus (CMV). A panel of recombinant modified
vaccinia Ankara (MVA) vectors, expressing various CMV
proteins (CMV-MVA) was used to immunize
HLA-A*0201, B*0702 and A*1101 Tg mice. Immune splenocytes were in vitro stimulated (IVS) either using syngeneic lipo-
polysaccharide activated lymphoblasts or Tg HLA-I matched human EBV-transformed B-lymphoblastoid cells (LCL), both loaded with
peptide libraries, encompassing the CMV
protein under investigation. IVS performed with
peptide library loaded lymphoblasts failed to provide a reliable stimulation. In contrast, the usage of LCL as antigen presenting cells (APC) of CMV
peptide libraries resulted in a consistent and specific amplification of the Tg T-cell response in animals immunized with CMV-MVAs. The LCL IVS method reliably allowed defining the immunogenicity and immunodominant CD8(+) T-cell regions of uncharacterized CMV
antigens. The combination of CMV-MVA vectors, unbiased pools of CMV-specific
peptide libraries presented by Tg HLA-I matched LCL constitutes a valid tool for the pre-clinical evaluation of model candidate
vaccines. This convenient method could find application to investigate the immunogenicity profile of
cancer antigens or
proteins from infectious human pathogens.