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. 2015 Oct 12;6:500. doi: 10.3389/fimmu.2015.00500

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Copyright © 2015 Bougherara, Mansuet-Lupo, Alifano, Ngô, Damotte, Le Frère-Belda, Donnadieu and Peranzoni.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Collagen fibers control the distribution and migration of CD8 T cells in the stroma of human ovarian carcinoma. (A) Human ovarian tumor slice stained for CD8 (green) and EpCAM (blue), showing accumulation of resident CD8 T cells in collagen-loose regions. Collagen (red) has been imaged through SHG. (B) Automatically counted number of resident CD8 T cells in different 50 μm × 50 μm stromal regions classified according to the surface (%) occupied by collagen detected through SHG. Results were obtained on slices from four different ovarian tumors. (C) Left panel: mean velocities of resident CD8 T cells in loose and dense collagen areas of individual human ovarian carcinomas. Right panel: pooled values, mean ± SD. (D–G) Motile behavior of resident CD8 T cells relative to collagen fibers. (D) Snapshots are shown at various time intervals for the same CD8 T cell. The trajectory of this CD8 T cell was superimposed over SHG images. The track is color-coded according to the instantaneous velocity. See also Movie S11 in Supplementary Material. (E) Velocity of the CD8 T cell represented in (D). (F) Snapshots are shown at various time intervals for the same CD8 T cell located in a linear collagen region. The trajectory of the CD8 T cell was superimposed over SHG images. The track is color-coded according to the instantaneous velocity. See also Movie S12 in Supplementary Material. (G) The graph shows the straightness index of individual tracks of CD8 T cells in linear and non-linear collagen fiber regions. Straightness indices close to 1 correspond to linear trajectories. Values are compiled from four different ovarian tumors. Mann–Whitney test, *P < 0.05; **P < 0.01; ***P < 0.001.