BACKGROUND AIMS: Chimeric antigen receptor (CAR) T cells are a novel immunotherapy for @entity5 . To achieve anti- @entity5 efficacy, these cells must survive, expand, and persist after infusion into @entity1 , functions that are reportedly best achieved by cells with a stem or central-memory rather than effector-memory phenotype. We have developed third-generation CAR T cells specific for the @entity5 -associated antigen GD2 for use in a phase I clinical trial. We investigated the optimal cell culture conditions for CAR T-cell production, and here we describe the relative effects of 3 activation and cytokine conditions on CAR T-cell expansion, effector function and phenotype. METHODS: Peripheral blood mononuclear cells were activated by anti-CD3 and anti- @entity1361 or anti-CD3 and Retronectin. Activated cells were transduced with the CAR-encoding retroviral vector and expanded in either interleukin (IL)-2 or @entity6977 and @entity8252 . Immune phenotype and expansion were tracked throughout the culture, and transduction efficiency, and subsequent GD2-specific effector functions were evaluated by flow cytometry and cytotoxic T lymphocytes assay. RESULTS: CD3/Retronectin stimulation with @entity1199 resulted in poorer activation, expansion and @entity1198 cytokine secretion of CAR T cells than CD3/ @entity1361 stimulation with either @entity1199 or @entity6977 and @entity8252 . However, CAR T cells cultured in CD3/ @entity1361 / @entity51148 and CD3/Retronectin/ @entity1199 had superior cytotoxic T lymphocyte activity and a more stem-like phenotype. DISCUSSION: The combination of CD3 and @entity1361 with @entity6977 and @entity8252 gave the best balance of CAR T-cell expansion and potent GD2-specific effector functions while retaining a stem/memory phenotype, and these growth conditions will therefore be used to manufacture CAR T cells for our phase I clinical trial.

Different cytokine and stimulation conditions influence the expansion and immune phenotype of third-generation chimeric antigen receptor T  cells specific for XXXX antigen GD2.

@entity1 :: ('9606', 'Species') :: ['patients'] @entity5 :: ('MESH:D009369', 'Disease') :: ['tumor', 'cancer'] @entity6977 :: ('3574', 'Gene') :: ['IL-7'] @entity1198 :: ('51497', 'Gene') :: ['Th1'] @entity1199 :: ('3558', 'Gene') :: ['IL-2'] @entity8252 :: ('3600', 'Gene') :: ['IL-15'] @entity51148 :: ('3574;3600', 'Gene') :: ['IL7/IL-15'] @entity1361 :: ('940', 'Gene') :: ['CD28']
@entity5 :: (MESH:D009369,Disease) :: ['tumor']
