Fig. 3: Immunological synapse formation between pAPCs and cognate T cells. | Nature Communications

Fig. 3: Immunological synapse formation between pAPCs and cognate T cells.

From: Polymerised superparamagnetic antigen presenting cell lymphocyte capture for enriching tumour reactive T-cells and neoantigen identification

Fig. 3

A Schematic illustration of polarized MHC-I and ICAM-1 distributions on APCs at the APC/T cell interface and fluorescence microscopy images showing patterned MHC-I and ICAM-1 distributions on pAPCs. The experiment was independently repeated two times. Scale bars = 10 and 4 μm. B Relative MHC fluorescence intensities and C ICAM-1 fluorescence intensities at the cellular contact interface and peripheral regions away from cellular contact are quantified. P value ****p < 0.0001 in (B) and ***p = 0.0005 in (C) (Independent biological replicates n = 5). D Schematic illustration of polarized TCR and LFA-1 distributions on T cells at the APC/T cell interface and fluorescence microscopy images showing patterned TCR and LFA-1 distributions on pAPC-engaged T cells. The experiment was independently repeated two times. Scale bars = 10 and 2 μm. E Relative TCR fluorescence intensities and F LFA-1 fluorescence intensities at the cellular contact interface and peripheral regions away from cellular contact are quantified. P value in ***p = 0.0006 in (E) and ****p < 0.0001 in (F) (Independent biological replicates n = 5). G OT-I CD8+ T cells engaging with SIINFEKL-loaded pAPCs exhibit F-actin polarization at the cell-cell interface. The experiment was independently repeated two times. Scale bars = 10 and 2 μm. H Relative phalloidin intensities at the cellular contact interface and peripheral regions away from cellular contact are quantified. P value ****p < 0.0001 (Independent biological replicates n = 5). I Time-lapse microscopy images showing magnetized pAPC carrying engaged T cells upon magnetically induced movement. Arrows indicate the movement direction of pAPCs. Scale bar = 10 μm. J Schematic illustration of imaging cytometry analysis of APC/T cell engagement and representative fluorescence microscopy images of APC/T cell engagement events taken after 2 h of incubation. White arrows indicate engagement events. The experiment was independently repeated six times. Scale bar = 40 µm. K Kinetic analysis of engagement events over a 3 h incubation period. P values from left to right, NS = 0.4530, ****p < 0.0001, ****p < 0.0001 (Independent biological replicates n = 6). The error bars represent mean ± standard deviation. Statistical analyses were performed by (B, C, E, F, H) two-sided Student’s t tests and (K) one-way ANOVA.

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