Tetramer guided, cell sorter assisted production of clinical grade autologous NY-ESO-1 specific CD8(+) T cells.

[liposarcoma]

Adoptive T cell therapy represents an attractive modality for the treatment of patients with cancer . Peripheral blood mononuclear cells have been used as a source of antigen specific T cells but the very low frequency of T cells recognizing commonly expressed antigens such as NY -ESO-1 limit the applicability of this approach to other solid tumors . To overcome this , we tested a strategy combining IL -21 modulation during in vitro stimulation with first -in -class use of tetramer-guided cell sorting to generate NY -ESO-1 specific cytotoxic T lymphocytes (CTL ).CTL generation was evaluated in 6 patients with NY -ESO-1 positive sarcomas , under clinical manufacturing conditions and characterized for phenotypic and functional properties .Following in vitro stimulation , T cells stained with NY -ESO-1 tetramer were enriched from frequencies as low as 0 .4 % to >90 % after single pass through a clinical grade sorter . NY -ESO-1 specific T cells were generated from all 6 patients . The final products expanded on average 1200 -fold to a total of 36 billion cells , were oligoclonal and contained 67 -97 % CD 8 (+), tetramer (+) T cells with a memory phenotype that recognized endogenous NY -ESO-1 .This study represents the first series using tetramer-guided cell sorting to generate T cells for adoptive therapy . This approach , when used to target more broadly expressed tumor antigens such as WT-1 and additional Cancer -Testis antigens will enhance the scope and feasibility of adoptive T cell therapy .