title: Tetramer guided, cell sorter assisted production of clinical grade autologous NY-ESO-1 specific CD8(+) T cells.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Department of Medicine, University of Washington, Seattle, WA USA.
authors.forename: Seth M
authors.initials: SM
authors.lastname: Pollack
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Department of Medicine, University of Washington, Seattle, WA USA.
authors.forename: Robin L
authors.initials: RL
authors.lastname: Jones
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA.
authors.forename: Erik A
authors.initials: EA
authors.lastname: Farrar
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Institute for Advanced Study, Technical University of Munich, Munich, Germany.
authors.forename: Ivy P
authors.initials: IP
authors.lastname: Lai
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Department of Medicine, University of Washington, Seattle, WA USA.
authors.forename: Sylvia M
authors.initials: SM
authors.lastname: Lee
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA.
authors.forename: Jianhong
authors.initials: J
authors.lastname: Cao
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Department of Surgery, University of Washington, Seattle, WA USA.
authors.forename: Venu G
authors.initials: VG
authors.lastname: Pillarisetty
authors.affiliation: Department of Pathology, University of Washington, Seattle, WA USA.
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Hoch
authors.affiliation: Department of Pathology, University of Washington, Seattle, WA USA.
authors.forename: Ashley
authors.initials: A
authors.lastname: Gullett
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Department of Pediatrics, University of Washington, Seattle, WA USA.
authors.forename: Marie
authors.initials: M
authors.lastname: Bleakley
authors.affiliation: Department of Orthopedics, University of Washington, Seattle, WA USA.
authors.forename: Ernest U
authors.initials: EU
authors.lastname: Conrad
authors.affiliation: Department of Radiology, University of Alabama, Birmingham, AL USA.
authors.forename: Janet F
authors.initials: JF
authors.lastname: Eary
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA.
authors.forename: Kendall C
authors.initials: KC
authors.lastname: Shibuya
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Department of Medicine, University of Washington, Seattle, WA USA.
authors.forename: Edus H
authors.initials: EH
authors.lastname: Warren
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA.
authors.forename: Jason N
authors.initials: JN
authors.lastname: Carstens
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA.
authors.forename: Shelly
authors.initials: S
authors.lastname: Heimfeld
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Department of Medicine, University of Washington, Seattle, WA USA ; Institute for Advanced Study, Technical University of Munich, Munich, Germany.
authors.forename: Stanley R
authors.initials: SR
authors.lastname: Riddell
authors.affiliation: Clinical Research Division, D3-100 Fred Hutchinson Cancer Research Center, 1100 Fairview Ave, Seattle, WA 98109 USA ; Department of Medicine, University of Washington, Seattle, WA USA ; Department of Melanoma Medical Oncology, UT MD Anderson Cancer Center, 7455 Fannin St, Unit 904, Houston, TX 77054 USA.
authors.forename: Cassian
authors.initials: C
authors.lastname: Yee
keywords: Adoptive T cell therapy; Antigen specific T cells; Immunotherapy; Influx cell sorting; Liposarcoma; Myxoid; NY-ESO-1; Synovial sarcoma; Tetramer