Efficient transfer of a tumor antigen-reactive TCR to human peripheral blood lymphocytes confers anti-tumor reactivity

The tumor-associated Ag MART-1 is expressed by most human melanomas. The ge nes encoding an alpha beta TCR from a MART-1-specific, HLA-A2-restricted, h uman T cell clone have been efficiently transferred and expressed in human PBL, These retrovirally transduced PBL cultures were MART-1 peptide reactiv e, and most cultures recognized HLA-A2(+) melanoma lines. Limiting dilution clones were generated from three bulk transduced PBL cultures to investiga te the function of individual clones within the transduced cultures. Twenty -nine of 29 CD8(+) clones specifically secreted IFN-gamma in response to T2 cells pulsed with MART-1((27-35)) peptide, and 23 of 29 specifically secre ted IFN-gamma in response to HLA-A2+ melanoma Lines. Additionally, 23 of 29 CD8+ clones lysed T2 cells pulsed with the MART-1((27-35)) peptide and 15 of 29 lysed the HLA-A2(+) melanoma line 888. CD4(+) clones specifically sec reted IFN-gamma in response to T2 cells pulsed with the MART-1((27-35)) pep tide. TCR gene transfer td patient PBL can produce CTL with anti-tumor reac tivity in vitro and could potentially offer a treatment for patients with m etastatic melanoma, This approach could also be applied to the treatment of other tumors and viral infections. Additionally, TCR gene transfer offers unique opportunities to study the fate of adoptively transferred T cells in vivo.