title: Chimeric antigen receptor-redirected T cells display multifunctional capacity and enhanced tumor-specific cytokine secretion upon secondary ligation of chimeric receptor.
abstract: OBJECTIVE
  The aim of the current study was to fully elucidate the functions of T cells genetically modified with an erbB2-specific chimeric antigen receptor (CAR).
  
  
  METHODS
  In this study, key functional parameters of CAR T cells were examined following antigen-specific stimulation of the chimeric anti-erbB2 receptor.
  
  
  RESULTS
  Gene-modified T cells produced the cytokines IFN-γ, IL-2, IL-4, IL-10, TNF-α and IL-17, and the chemokine RANTES upon CAR ligation. A multifunctional capacity of these CAR T cells was also demonstrated, where 13.7% of cells were found to simultaneously express IFN-γ and CD107a, indicative of cytolytic granule release. In addition, the CAR T cells were able to respond to a greater degree on the second ligation of CAR, which has not been previously shown. IFN-γ secretion levels were significantly higher on second ligation than those secreted following initial ligation. CAR-expressing T cells were also demonstrated to lyze erbB2-expressing tumor cells in the absence of activity against bystander cells not expressing the erbB2 antigen, thereby demonstrating exquisite specificity.
  
  
  CONCLUSIONS
  This study demonstrates the specificity of CAR gene-engineered T cells and their capacity to deliver a wide range of functions against tumor cells with an enhanced response capability after initial receptor engagement.
authors.affiliation: Cancer Immunology Research Program, Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Henderson
authors.forename: Carmen S M
authors.initials: CS
authors.lastname: Yong
authors.forename: Connie P M
authors.initials: CP
authors.lastname: Duong
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Davenport
authors.forename: Liza B
authors.initials: LB
authors.lastname: John
authors.forename: Christel
authors.initials: C
authors.lastname: Devaud
authors.forename: Paul
authors.initials: P
authors.lastname: Neeson
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Westwood
authors.forename: Phillip K
authors.initials: PK
authors.lastname: Darcy
authors.forename: Michael H
authors.initials: MH
authors.lastname: Kershaw
mesh_terms: D000818:Animals; D018106:CD28 Antigens / Q000235:genetics / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D045325:HCT116 Cells; D006801:Humans; D007156:Immunologic Memory; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008213:Lymphocyte Activation / Q000235:genetics; D051379:Mice; D009369:Neoplasms / Q000628:therapy*; D015202:Protein Engineering; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology / Q000637:transplantation*
chemical_list: D018106:CD28 Antigens; D016207:Cytokines; D018719:Erb-b2 Receptor Tyrosine Kinases; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; C508053:ERBB2 protein, human