title: A novel approach to evaluate the immunogenicity of viral antigens of clinical importance in HLA transgenic murine models.
abstract: 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.
authors.affiliation: Laboratory of Vaccine Research, Beckman Research Institute of the City of Hope, City of Hope Comprehensive Cancer Center, Duarte, CA 91010, USA.
authors.forename: Aparna
authors.initials: A
authors.lastname: Krishnan
authors.forename: Zhongde
authors.initials: Z
authors.lastname: Wang
authors.forename: Tumul
authors.initials: T
authors.lastname: Srivastava
authors.forename: Ravindra
authors.initials: R
authors.lastname: Rawal
authors.forename: Pooja
authors.initials: P
authors.lastname: Manchanda
authors.forename: Don J
authors.initials: DJ
authors.lastname: Diamond
authors.forename: Corinna
authors.initials: C
authors.lastname: La Rosa
mesh_terms: D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003587:Cytomegalovirus / Q000276:immunology; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D006680:HLA Antigens / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D017874:Immediate-Early Proteins / Q000276:immunology; D051379:Mice; D018345:Mice, Knockout; D010750:Phosphoproteins / Q000276:immunology; D015534:Trans-Activators / Q000276:immunology; D014764:Viral Proteins / Q000276:immunology; D014765:Viral Vaccines / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte; D006680:HLA Antigens; C069186:IE2 protein, Cytomegalovirus; D017874:Immediate-Early Proteins; D010750:Phosphoproteins; D015534:Trans-Activators; D014764:Viral Proteins; D014765:Viral Vaccines; C070746:human cytomegalovirus protein pp28