title: Antigen-specific in vitro expansion of functional redirected NY-ESO-1-specific human CD8+ T-cells in a cell-free system.
abstract: BACKGROUND
  Tumors can be targeted by the adoptive transfer of chimeric antigen receptor (CAR) redirected T-cells. Antigen-specific expansion protocols are needed to generate large quantities of redirected T-cells. We aimed to establish a protocol to expand functional active NY-ESO-1-specific redirected human CD8(+) T-cells.
  
  
  METHODS
  The anti-idiotypic Fab antibody A4 with specificity for HLA-A 0201/NY-ESO-1157-165 was tested by competition assays using a HLA-A 0201/NY-ESO-1157-165 tetramer. HLA-A 0201/NY-ESO-1157-165 redirected T-cells were generated, expanded and tested for CAR expression, cytokine release, in vitro cytolysis and protection against xenografted HLA-A 0201/NY-ESO-1157-165-positive multiple myeloma cells.
  
  
  RESULTS
  A4 demonstrated antigen-specific binding to HLA-A 0201/NY-ESO-1157-165 redirected T-cells. Expansion with A4 resulted in 98% of HLA-A 0201/NY-ESO-1157-165 redirected T-cells. A4 induced strong proliferation, resulting in a 300-fold increase of redirected T-cells. After expansion protocols, redirected T-cells secreted Interleukin-2, (IL-2), interferon gamma (IFNγ) and tumor necrosis factor alpha (TNFα) and lysed target cells in vitro and were protective in vivo.
  
  
  CONCLUSIONS
  A4 expanded HLA-A 0201/NY-ESO-1157-165 redirected T-cells with preservation of antigen-specific function.
authors.affiliation: Clinic of Oncology, University Hospital, Zurich, Rämistr. 100, 8091 Zürich, Switzerland. Ulf.Petrausch@usz.ch.
authors.forename: Gopinadh
authors.initials: G
authors.lastname: Jakka
authors.forename: Petra C
authors.initials: PC
authors.lastname: Schuberth
authors.forename: Markus
authors.initials: M
authors.lastname: Thiel
authors.forename: Gerhard
authors.initials: G
authors.lastname: Held
authors.forename: Frank
authors.initials: F
authors.lastname: Stenner
authors.forename: Maries
authors.initials: M
authors.lastname: Van Den Broek
authors.forename: Christoph
authors.initials: C
authors.lastname: Renner
authors.forename: Axel
authors.initials: A
authors.lastname: Mischo
authors.forename: Ulf
authors.initials: U
authors.lastname: Petrausch
mesh_terms: D000818:Animals; D000915:Antibody Affinity; D000951:Antigens, Neoplasm / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000637:transplantation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002474:Cell-Free System; D018920:Coculture Techniques; D003602:Cytotoxicity, Immunologic; D057809:HEK293 Cells; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D016219:Immunotherapy, Adoptive; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000276:immunology*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009101:Multiple Myeloma / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D009368:Neoplasm Transplantation; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D047368:Tumor Burden
chemical_list: D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D007140:Immunoglobulin Fab Fragments; D008565:Membrane Proteins
keywords: NY-ESO-1; Redirected T-cells; anti-idiotypic antibody; chimeric antigen receptor; expansion; peptide specific