title: Strong and sustained effector function of memory- versus naïve-derived T cells upon T-cell receptor RNA transfer: implications for cellular therapy.
abstract: Current protocols used to select CMV-specific T cells for adoptive immunotherapy focus on virus-specific memory T cells from seropositive donors. However, this strategy is not feasible in patients undergoing allogeneic haematopoietic stem-cell transplantation (HSCT) from CMV-seronegative donors. Here, we redirected T cells of CMV-seronegative donors with a human genetically engineered TCR recognizing an HLA-A*0201-binding peptide epitope of CMVpp65. To facilitate clinical translation of this approach, we used a non-viral expression system based on in vitro transcribed RNA and electroporation. Although memory and naïve-derived T-cell subsets were both efficiently transfected by TCR-RNA, memory-derived T cells showed much stronger levels of HLA-A*0201-restricted cytolytic activity to CMV-infected fibroblasts and maintained acquired function for 5-10 days. In addition to redirection of CD8(+)  cytotoxic T cells, TCR-RNA transfection was capable of redirecting CD4(+)  T cells into potent Ag-specific Th cells that efficiently triggered maturation of DCs. Our data suggest that memory rather than naïve-derived T cells are the preferred subset for transient TCR expression by RNA electroporation, providing more efficient and sustained virus-specific CD4(+)  and CD8(+)  T-cell function. CMV TCR-RNA may represent a suitable therapeutic 'off-the-shelf' reagent to be used in severe CMV infections of HSCT patients when endogenous CMV-specific T-cell immunity is insufficient.
authors.affiliation: Department of Medicine III, University Medical Center of Johannes Gutenberg-University Mainz, Mainz, Germany. thomass@uni-mainz.de
authors.forename: Simone
authors.initials: S
authors.lastname: Thomas
authors.forename: Sebastian
authors.initials: S
authors.lastname: Klobuch
authors.forename: Katrin
authors.initials: K
authors.lastname: Besold
authors.forename: Bodo
authors.initials: B
authors.lastname: Plachter
authors.forename: Jan
authors.initials: J
authors.lastname: Dörrie
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
authors.forename: Matthias
authors.initials: M
authors.lastname: Theobald
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Herr
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D064987:Cell- and Tissue-Based Therapy; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000276:immunology / Q000628:therapy; D018274:Electroporation / Q000379:methods; D005347:Fibroblasts / Q000276:immunology; D005786:Gene Expression Regulation / Q000235:genetics / Q000276:immunology; D006086:Graft vs Host Disease / Q000235:genetics / Q000276:immunology / Q000628:therapy; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D007156:Immunologic Memory / Q000187:drug effects* / Q000235:genetics; D010750:Phosphoproteins / Q000235:genetics / Q000276:immunology*; D012313:RNA / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology*; D014162:Transfection / Q000379:methods; D014184:Transplantation, Homologous; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D010750:Phosphoproteins; D011948:Receptors, Antigen, T-Cell; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D012313:RNA