title: Individual Motile CD4(+) T Cells Can Participate in Efficient Multikilling through Conjugation to Multiple Tumor Cells.
abstract: T cells genetically modified to express a CD19-specific chimeric antigen receptor (CAR) for the investigational treatment of B-cell malignancies comprise a heterogeneous population, and their ability to persist and participate in serial killing of tumor cells is a predictor of therapeutic success. We implemented Timelapse Imaging Microscopy in Nanowell Grids (TIMING) to provide direct evidence that CD4(+)CAR(+) T cells (CAR4 cells) can engage in multikilling via simultaneous conjugation to multiple tumor cells. Comparisons of the CAR4 cells and CD8(+)CAR(+) T cells (CAR8 cells) demonstrate that, although CAR4 cells can participate in killing and multikilling, they do so at slower rates, likely due to the lower granzyme B content. Significantly, in both sets of T cells, a minor subpopulation of individual T cells identified by their high motility demonstrated efficient killing of single tumor cells. A comparison of the multikiller and single-killer CAR(+) T cells revealed that the propensity and kinetics of T-cell apoptosis were modulated by the number of functional conjugations. T cells underwent rapid apoptosis, and at higher frequencies, when conjugated to single tumor cells in isolation, and this effect was more pronounced on CAR8 cells. Our results suggest that the ability of CAR(+) T cells to participate in multikilling should be evaluated in the context of their ability to resist activation-induced cell death. We anticipate that TIMING may be used to rapidly determine the potency of T-cell populations and may facilitate the design and manufacture of next-generation CAR(+) T cells with improved efficacy.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas.
authors.forename: Ivan
authors.initials: I
authors.lastname: Liadi
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Harjeet
authors.initials: H
authors.lastname: Singh
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas.
authors.forename: Gabrielle
authors.initials: G
authors.lastname: Romain
authors.affiliation: Department of Electrical Engineering, University of Houston, Houston, Texas.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Rey-Villamizar
authors.affiliation: Department of Electrical Engineering, University of Houston, Houston, Texas.
authors.forename: Amine
authors.initials: A
authors.lastname: Merouane
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas.
authors.forename: Jay R T
authors.initials: JR
authors.lastname: Adolacion
authors.affiliation: Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Partow
authors.initials: P
authors.lastname: Kebriaei
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Helen
authors.initials: H
authors.lastname: Huls
authors.affiliation: Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia.
authors.forename: Peng
authors.initials: P
authors.lastname: Qiu
authors.affiliation: Department of Electrical Engineering, University of Houston, Houston, Texas.
authors.forename: Badrinath
authors.initials: B
authors.lastname: Roysam
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas.
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Houston, Houston, Texas. nvaradar@central.uh.edu.
authors.forename: Navin
authors.initials: N
authors.lastname: Varadarajan
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D045744:Cell Line, Tumor; D053804:Granzymes / Q000276:immunology; D006801:Humans; D009369:Neoplasms / Q000276:immunology; D011946:Receptors, Antigen / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D011946:Receptors, Antigen; D053804:Granzymes