Reprogramming T lymphocytes for melanoma adoptive immunotherapy by T-cell receptor gene transfer with lentiviral vectors.
T-cell receptor (TCR) gene transfer for cancer immunotherapy is limited by the availability of large numbers of tumor-specific T cells. TCR alpha and beta chains were isolated from a highly lytic HLA-A2-restricted cytotoxic T lymphocyte (CTL) clone recognizing the melanoma-associated Melan-A/MART-1 antigen and inserted into a lentiviral vector carrying a bidirectional promoter capable of robust and coordinated expression of the two transgenes. Lentiviral vector-based gene delivery systems have shown increased transfer efficiency and transgene expression compared with the widely used gamma-retroviral vectors. This vector performed more efficiently than a gamma-retrovirus-based vector containing the same expression cassette, resulting in a T-cell population with 60% to 80% of transgenic TCR expression with mainly CD8(+) intermediate effector phenotype. Transgenic T cells specifically produced cytokine in response to and killed antigen-expressing melanoma cells, retained an overlapping functional avidity in comparison with the TCR donor CTL clone, and exerted significant therapeutic effects in vivo upon adoptive transfer in melanoma-bearing severe combined immunodeficient mice. Optical imaging showed their accumulation in the tumor site. Overall, our results indicate that lentiviral vectors represent a valid tool for stable and high-intensity expression of transgenic TCR and support clinical exploitation of this approach for therapeutic application.
Bobisse
Sara
S
Department of Oncology, University of Padova, Padova, Italy.
Rondina
Maria
M
Merlo
Anna
A
Tisato
Veronica
V
Mandruzzato
Susanna
S
Amendola
Mario
M
Naldini
Luigi
L
Willemsen
Ralph A
RA
Debets
Reno
R
Zanovello
Paola
P
Rosato
Antonio
A
D000818:Animals; D000951:Antigens, Neoplasm / Q000276:immunology; D000939:Epitopes; D005260:Female; D019673:Genes, T-Cell Receptor alpha; D019674:Genes, T-Cell Receptor beta; D005822:Genetic Vectors / Q000235:genetics; D015789:HLA-A2 Antigen / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory; D016219:Immunotherapy, Adoptive / Q000379:methods*; D019169:Jurkat Cells; D016086:Lentivirus / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000276:immunology; D058965:MART-1 Antigen; D008545:Melanoma / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D051379:Mice; D016513:Mice, SCID; D009363:Neoplasm Proteins / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000096:biosynthesis / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology / Q000502:physiology*; D014161:Transduction, Genetic
D000951:Antigens, Neoplasm; D000939:Epitopes; D015789:HLA-A2 Antigen; D058965:MART-1 Antigen; C548667:MLANA protein, human; C548668:Mlana protein, mouse; D009363:Neoplasm Proteins; D016693:Receptors, Antigen, T-Cell, alpha-beta