title: Generation of HIV-1-specific T cells by electroporation of T-cell receptor RNA.
abstract: BACKGROUND
  HIV-1-specific cytotoxic T lymphocytes, which recognize conserved epitopes of the virus, are correlated with prolonged survival of infected individuals. Unfortunately, most HIV-1-infected patients are unable to generate such an immune response. Antigen-specific cytotoxic T lymphocytes can be generated by T-cell receptor transfer. This is commonly done by retroviral transduction, which is complicated and poses the threat of stable genetic alteration of autologous cells.
  
  
  METHODS
  We reprogrammed primary CD8+ T cells by electroporation of RNA, which encoded an HIV-1-pol- and an HIV-1-gag-specific T-cell receptor recognizing the human leukocyte antigen-A2 restricted epitopes ILKEPVHGV and SLYNTVATL, respectively.
  
  
  RESULTS
  These reprogrammed cells specifically produced the proinflammatory cytokines interleukin-2, tumor necrosis factor-alpha, and interferon-gamma after stimulation with target cells that presented the corresponding peptides, and were able to lyse these targets efficiently and specifically. The lytic avidities of the HIV-1-pol- and HIV-1-gag-TCR-RNA-electroporated CD8+ T cells were within the same range than those of the parental cytotoxic T lymphocytes. Most importantly, HIV-1-gag-reprogrammed T cells recognized target cells that presented endogenously processed antigen, which resulted in cytokine production and lysis.
  
  
  CONCLUSIONS
  It is shown here for the first time that functional transfer of virus-specific T-cell receptors by RNA electroporation is feasible, and represents an innovative, safe, and easy method to generate virus-specific T cells, avoiding the risks of retroviral transduction.
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Erlangen, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Hofmann
authors.forename: Thomas
authors.initials: T
authors.lastname: Harrer
authors.forename: Verena
authors.initials: V
authors.lastname: Kubesch
authors.forename: Katja
authors.initials: K
authors.lastname: Maurer
authors.forename: Karin J
authors.initials: KJ
authors.lastname: Metzner
authors.forename: Kathrin
authors.initials: K
authors.lastname: Eismann
authors.forename: Silke
authors.initials: S
authors.lastname: Bergmann
authors.forename: Matthias
authors.initials: M
authors.lastname: Schmitt-Haendle
authors.forename: Gerold
authors.initials: G
authors.lastname: Schuler
authors.forename: Jan
authors.initials: J
authors.lastname: Dörrie
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003001:Cloning, Molecular; D016207:Cytokines / Q000032:analysis; D003601:Cytotoxicity Tests, Immunologic; D018274:Electroporation; D000939:Epitopes / Q000276:immunology; D015658:HIV Infections / Q000276:immunology* / Q000628:therapy*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007167:Immunotherapy; D012313:RNA / Q000494:pharmacology*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014158:Transcription, Genetic
chemical_list: D016207:Cytokines; D000939:Epitopes; D011948:Receptors, Antigen, T-Cell; D012313:RNA