title: T cell receptor (TCR) gene transfer with lentiviral vectors allows efficient redirection of tumor specificity in naive and memory T cells without prior stimulation of endogenous TCR.
abstract: We investigated the possibility of introducing exogenous T cell receptor (TCR) genes into T cells by lentiviral transduction, without prior stimulation of endogenous TCR with anti-CD3. TCR transfer is used to impose tumor antigen specificity on recipient T cells, but sustained activation required for retroviral transduction may affect the clinical efficacy of engineered T cells. Cytokine stimulation makes T cells susceptible to lentiviral transduction in the absence of TCR triggering, but this advantage has never been exploited for TCR transfer. Autoimmune diseases are a source of high-affinity TCRs specific for self/tumor antigens. We selected, from a patient with vitiligo, a Mart1-specific TCR based on intrinsic interchain pairing properties and functional avidity. After lentiviral transduction of human peripheral blood mononuclear cells, preferential pairing of exogenous alpha and beta chains was observed, together with effective recognition of Mart1(+) melanoma cells. We tested transduction efficiency on various T cell subsets prestimulated with interleukin (IL)-2, IL-7, IL-15, and IL-21 (alone or in combination). Both naive and unfractionated CD8(+) T cells could be transduced without requiring endogenous TCR triggering. IL-7 plus IL-15 was the most powerful combination, allowing high levels of transgene expression without inducing T cell differentiation (34 +/- 5% Mart1-TCR(+) cells in naive CD8(+) and 16 +/- 6% in unfractionated CD8(+)). Cytokine-prestimulated, Mart1-redirected naive and unfractionated CD8(+) cells expanded better than CD3-CD28-prestimulated counterparts in response to both peptide-pulsed antigen-presenting cells and Mart1(+) melanoma cells. This strategy allows the generation of tumor-specific T cells encompassing truly naive T cells, endowed with an intact proliferative potential and a preserved differentiation stage.
authors.affiliation: Laboratory of Cancer Immunology, Institute for Cancer Research and Treatment (IRCC), Candiolo, 10060, Turin, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Circosta
authors.forename: Luisa
authors.initials: L
authors.lastname: Granziero
authors.forename: Antonia
authors.initials: A
authors.lastname: Follenzi
authors.forename: Elisa
authors.initials: E
authors.lastname: Vigna
authors.forename: Stefania
authors.initials: S
authors.lastname: Stella
authors.forename: Antonella
authors.initials: A
authors.lastname: Vallario
authors.forename: Angela Rita
authors.initials: AR
authors.lastname: Elia
authors.forename: Loretta
authors.initials: L
authors.lastname: Gammaitoni
authors.forename: Katiuscia
authors.initials: K
authors.lastname: Vitaggio
authors.forename: Francesca
authors.initials: F
authors.lastname: Orso
authors.forename: Massimo
authors.initials: M
authors.lastname: Geuna
authors.forename: Dario
authors.initials: D
authors.lastname: Sangiolo
authors.forename: Maja
authors.initials: M
authors.lastname: Todorovic
authors.forename: Claudia
authors.initials: C
authors.lastname: Giachino
authors.forename: Alessandro
authors.initials: A
authors.lastname: Cignetti
mesh_terms: D000951:Antigens, Neoplasm / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D045744:Cell Line, Tumor; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007156:Immunologic Memory; D016086:Lentivirus / Q000235:genetics; D058965:MART-1 Antigen; D008545:Melanoma / Q000276:immunology / Q000628:therapy*; D009363:Neoplasm Proteins / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D014161:Transduction, Genetic; D014820:Vitiligo / Q000276:immunology
chemical_list: D000951:Antigens, Neoplasm; D058965:MART-1 Antigen; C548667:MLANA protein, human; D009363:Neoplasm Proteins; D011948:Receptors, Antigen, T-Cell