Generation of HIV-1-specific T cells by electroporation of T-cell receptor RNA.
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.
Hofmann
Christian
C
Department of Dermatology, University Hospital Erlangen, Erlangen, Germany.
Harrer
Thomas
T
Kubesch
Verena
V
Maurer
Katja
K
Metzner
Karin J
KJ
Eismann
Kathrin
K
Bergmann
Silke
S
Schmitt-Haendle
Matthias
M
Schuler
Gerold
G
Dörrie
Jan
J
Schaft
Niels
N
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
D016207:Cytokines; D000939:Epitopes; D011948:Receptors, Antigen, T-Cell; D012313:RNA