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. 2022 Nov 4;1(5):pgac249. doi: 10.1093/pnasnexus/pgac249

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© The Author(s) 2022. Published by Oxford University Press on behalf of National Academy of Sciences.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Fig. 3. — Acute mucosal wounds induce PMN swarming. (A and B) Rodent colonoscope and representative image of colonic biopsy-based wound. (C) Representative flow cytometry plot showing transmigrated PMNs (CD45⁺Ly6G⁺CD11b^high) in wounded mucosa compared to intact tissue. (D and E) Quantification of wound-associated PMNs after biopsy-induced colonic injury (0 to 24 h). (D) Graph represents number of PMNs per 2.5 × 10⁵ cells of lamina propria cells. Data are means ± SEM of two independent experiments (n = 5 to 7 mice per group; ***P < 0.001; ^****P < 0.0001 compared to Intact tissue by one-way ANOVA). (E) Represents percentage of wound associated GFP⁺ cells. Data are means ± SEM of two independent experiments (n = 5 to 7 mice per group; *P < 0.05; ***P < 0.001; ^****P < 0.0001 compared to Intact tissue by two-way ANOVA. Asterisk colors represent compared groups matching figure keys). (F) Schematic overview of biopsy-wounding model: GFP^high PMNs swarming towards focalized, acute mucosal ulcer, characteristic of ulcerative colitis. (G) Representative image of GFP^high PMNs swarming towards a colonic biopsy wound (white-dashed line) (green = GFP, red = vasculature, white = EpCAM). White-dashed line limits the wounded area. Scale bar: 200 μm. (H) Representative image of PMNs interacting with intestinal epithelial cells (green = GFP, red = vasculature, white = EpCAM). Scale bar: 20 μm. (I and J) Comparison of PMN dynamics on DSS-treated and LysM^eGFP/+vs. Cd47⁻^/⁻; LysM^eGFP/+ wounded mice (tracks of at least 20 GFP^high PMN per mouse; n = 3 to 6 mice per group). (I) Mean square displacement and (J) Mean track speed. Data are represented as median (interquartile range). ^****P < 0.0001 as determined by Wilcoxson–Mann–Whitney nonparametric test. (K) Representative images of PMN swarming towards a colonic mucosal wound on Cd47⁻^/⁻; LysM^eGFP/+mice (green: GFP, red: vasculature, white: EpCAM). White-dashed line limits wounded area. Asterisk points to migrating PMNs into wound. Scale bar: 100 μm. (L) Comparison of PMN track straightness on LysM^eGFP/+vs. Cd47⁻^/⁻; LysM^eGFP/+wounded mice. Tracks of at least 20 GFP^high PMNs per mouse; n = 3 to 6 mice per group. Data are represented as median (interquartile range). ^****P < 0.0001 as determined by Wilcoxson–Mann–Whitney nonparametric test.