title: B cells immortalized by a mini-Epstein-Barr virus encoding a foreign antigen efficiently reactivate specific cytotoxic T cells.
abstract: Lymphoblastoid cell lines (LCLs) are human B cells latently infected and immortalized by Epstein-Barr virus (EBV). Presenting viral antigens, they efficiently induce EBV-specific T-cell responses in vitro. Analogous ways to generate T-cell cultures specific for other antigens of interest are highly desirable. Previously, we constructed a mini-EBV plasmid that consists of less than half the EBV genome, is unable to cause virus production, but still immortalizes B cells in vitro. Mini-EBV-immortalized B-cell lines (mini-LCLs) are efficiently produced by infection of B cells with viruslike particles carrying only mini-EBV DNA. Mini-EBV plasmids can be engineered to express an additional gene in immortalized B cells. Here we present a mini-EBV coding for a potent CD8(+) T-cell antigen, the matrix phosphoprotein pp65 of human cytomegalovirus (CMV). By means of this pp65 mini-EBV, pp65-expressing mini-LCLs could be readily established from healthy donors in a one-step procedure. We used these pp65 mini-LCLs to reactivate and expand effector T cells from autologous peripheral blood cells in vitro. When generated from cytomegalovirus (CMV)-seropositive donors, these effector T-cell cultures displayed strong pp65-specific HLA-restricted cytotoxicity. A large fraction of CD8(+) T cells with pp65 epitope specificity was present in such cultures, as demonstrated by direct staining with HLA/peptide tetramers. We conclude that the pp65 mini-EBV is an attractive tool for CMV-specific adoptive immunotherapy. Mini-EBVs could also facilitate the generation of T cells specific for various other antigens of interest.
authors.affiliation: Department of Otorhinolaryngology, Ludwig-Maximilians-Universität, Munich, Germany. moosmann@hno.med.uni-muenchen.de
authors.forename: Andreas
authors.initials: A
authors.lastname: Moosmann
authors.forename: Naeem
authors.initials: N
authors.lastname: Khan
authors.forename: Mark
authors.initials: M
authors.lastname: Cobbold
authors.forename: Caroline
authors.initials: C
authors.lastname: Zentz
authors.forename: Henri-Jacques
authors.initials: HJ
authors.lastname: Delecluse
authors.forename: Gabi
authors.initials: G
authors.lastname: Hollweck
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Hislop
authors.forename: Neil W
authors.initials: NW
authors.lastname: Blake
authors.forename: Debbie
authors.initials: D
authors.lastname: Croom-Carter
authors.forename: Barbara
authors.initials: B
authors.lastname: Wollenberg
authors.forename: Paul A H
authors.initials: PA
authors.lastname: Moss
authors.forename: Reinhard
authors.initials: R
authors.lastname: Zeidler
authors.forename: Alan B
authors.initials: AB
authors.lastname: Rickinson
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hammerschmidt
mesh_terms: D000956:Antigens, Viral / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology* / Q000473:pathology / Q000821:virology*; D002472:Cell Transformation, Viral / Q000276:immunology*; D002478:Cells, Cultured; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D008213:Lymphocyte Activation; D008211:Lymphocyte Cooperation / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D018984:Epitopes, T-Lymphocyte