Article Title: Multidimensional single-cell analysis identifies a role of CD2-CD58 interactions for clinical antitumor T cell responses
Figure Lengend Snippet: (a) Schematic overview of the dynamic profiling and image analysis workflow of CAR T cell polyfunctionality using TIMING. We evaluated the interaction between CAR T cells and NALM-6 cells as tumor cells on arrays of nanowells using TIMING. (b) A representative example of a polyfunctional 19-28z T cell that participated in killing and secreted IFN-γ. TIMING is utilized to quantify T-cell intrinsic behavior like directional migration and the kinetics of the interaction leading to induction of apoptosis within tumor cells. After the TIMING assay, the IFN-γ molecules captured onto the beads during TIMING are revealed by using appropriate fluorescently labeled antibodies. Scale bar is 20 μm. (c) Schematic description of kinetic parameters measured in TIMING experiments. (d/g) Cumulative contact duration between effector and tumor cells leading to different functional outcomes. Effector cells that only secrete IFN-γ (monofunctional) exhibited longer contact duration compared to cytolytic cells with or without IFN-γ secretion. The data is aggregated from profiling all five IPs, the bar represents the median and the dotted lines quartiles. (e/h) Comparative assessments of t Contact and t Death for all killer 19-28z T cells. (f/i) Out-of-contact migration of the different functional subsets of 19-28z T cells. All data in (d)-(f) corresponds to E:T of 1:1 and is aggregated from profiling all five IPs (1589 T cells). All data in (g)-(i) corresponds to an E:T of 1:2-5 (to evaluate serial killing) [1178 T cells]. All p values for all multiple comparisons were computed using Kruskal-Wallis non-parametric tests and pairwise comparisons using a Mann-Whitney test. The black bar represents the median and the dotted lines denote quartiles.
Article Snippet: Human pre-B cell leukemic line NALM-6 (ATCC) were cultured in T-cell medium (RPMI + 10% FBS) and used as CD19 + target cells.
Techniques: Migration, Labeling, Functional Assay, MANN-WHITNEY