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. 2018 Sep 20;3(18):e120757. doi: 10.1172/jci.insight.120757

Figure 8. Bona fide IK are more abundant in malignant areas of human GBMs.

Figure 8

(A) Three-dimensional rendering of a confocal scanning of the tissue block biopsy containing T cells. Higher magnification of the reconstructed transparency shows a representative CD3+ T cell with kinaptic morphology (green). Zenithal view of the kinaptic T cell reveals the characteristic triangular shape with a clear leading lamellipodium (arrow) and TCR/CD3-rich uropod (arrowhead). The visualization of the GFAP+ adjacent tumor cells (magenta) and DAPI+ nuclei (blue) revealed close apposition with malignant cells. Zoom-in image of the transparency-based reconstruction displays the shape and details of the CD3 distribution. Magnification, 2×. An illustration depicting the interpretation of the T cell shape is also shown. (B) Confocal transparency projection shows the distribution of CD3 with higher fluorescence intensity at the trailing uropod. Fluorescence intensity scale (FI-CD3) evidences the uropod highly fluorescent. (C) Measurement of the relative fluorescence of CD3 along the yellow broken line revealed a scarce increase of fluorescence at the front lamellipodium (arrow 1) but a high increase at the uropod (arrow 2). (D) Three-dimensional isosurface rendering of kinaptic cell distinguishing the areas with low and high CD3 intensity and nucleus. Zenithal view, slight rotation of the x axis, and slight rotation of the z axis are shown. Scale bar: 2 μm. (E) Central optical plane of the T cell kinapse shows the orientation of the nuclear indentation toward the uropod, suggesting the direction of the motile T cell (blue arrow). A diagram for interpretation is also shown. (F) Examples of T cells with either regular rounded or triangular shape, and the latter with a high fluorescent CD3+ uropod. On the outline, red circles limit the leading front, and green circle indicates the CD3-rich uropod of the kinaptic T cell. Graphs show the quantification of percentages of T cells with triangular shape and with bona fide IK. ***P < 0.001 and ****P < 0.0001, Student t test and Mann-Whitney U test. (G) Illustrative diagram of the shape of a human T cell kinapse in a malignant area.