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

Seth M. Pollack*, Robin L. Jones, Erik A. Farrar, Ivy P. Lai, Sylvia M. Lee, Jianhong Cao, Venu G. Pillarisetty, Benjamin L. Hoch, Ashley Gullett, Marie Bleakley, Ernest U. Conrad, Janet F. Eary, Kendall C. Shibuya, Edus H. Warren, Jason N. Carstens, Shelly Heimfeld, Stanley R. Riddell, Cassian Yee

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

47 Scopus citations


Background: 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).Methods: CTL generation was evaluated in 6 patients with NY-ESO-1 positive sarcomas, under clinical manufacturing conditions and characterized for phenotypic and functional properties.Results: 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% CD8+, tetramer+ T cells with a memory phenotype that recognized endogenous NY-ESO-1.Conclusion: 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.

Original languageEnglish (US)
Article number36
JournalJournal for immunotherapy of cancer
Issue number1
StatePublished - Oct 14 2014
Externally publishedYes


  • Adoptive T cell therapy
  • Antigen specific T cells
  • Immunotherapy
  • Influx cell sorting
  • Liposarcoma
  • Myxoid
  • NY-ESO-1
  • Synovial sarcoma
  • Tetramer

ASJC Scopus subject areas

  • Molecular Medicine
  • Oncology
  • Cancer Research
  • Immunology and Allergy
  • Pharmacology
  • Immunology


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