TY - JOUR
T1 - 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
AU - Circosta, Paola
AU - Granziero, Luisa
AU - Follenzi, Antonia
AU - Vigna, Elisa
AU - Stella, Stefania
AU - Vallario, Antonella
AU - Elia, Angela Rita
AU - Gammaitoni, Loretta
AU - Vitaggio, Katiuscia
AU - Orso, Francesca
AU - Geuna, Massimo
AU - Sangiolo, Dario
AU - Todorovic, Maja
AU - Giachino, Claudia
AU - Cignetti, Alessandro
PY - 2009/12/1
Y1 - 2009/12/1
N2 - 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 α and β 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.
AB - 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 α and β 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.
UR - http://www.scopus.com/inward/record.url?scp=70349929446&partnerID=8YFLogxK
U2 - 10.1089/hum.2009.117
DO - 10.1089/hum.2009.117
M3 - Article
SN - 1043-0342
VL - 20
SP - 1576
EP - 1588
JO - Human Gene Therapy
JF - Human Gene Therapy
IS - 12
ER -