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. 2022 Jul 27;13:953738. doi: 10.3389/fmicb.2022.953738

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Copyright © 2022 Rimondi, Olivera, Soria, Parisi, Rumbo and Perez.

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) and the copyright owner(s) 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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Differences in the recruitment of mononuclear cells and/or lymphocyte subpopulations in ileum of IAV-infected chickens. Six animals per group were euthanized at 3 dpi after infection with IAV or inoculation with PBS at 3 weeks of age. Ileum was collected and lamina propria and epithelial compartments were processed for flow cytometry analyses. (A) Mononuclear cell recovery from epithelial (IEL) and lamina propria (LP) compartment. Results are expressed as the mean (SD) of total amount of cells/ml recovered from each compartment from ten cm of ileum. p-values were determined using ANOVA with multiple testing (Dunn’s Test for IEL and Tukey’s Test for LP). (B) Chicken infection with 20Ch557/H6N2 virus increases TCRγδ⁺ IELs from ileum. Cells were labeled with anti-CD3, anti-CD8α and anti-TCRγδ antibodies conjugated to SPRD, FITC and PE, respectively. Analysis was performed in the gate of lymphocytes according to forward/side scatter parameters, selecting CD3⁺ cells followed by TCRγδ-PE/CD8α-FICT. The results obtained in epithelial compartment analysis are shown, expressed as the mean in percentage (SD) from each group. p-values were determined using ANOVA with multiple testing (Tukey’s Test). (C) Macrophages in lamina propria from ileum of chickens infected with 20Ch557/H6N2 virus. Cells were labeled with anti-KUL01 and anti-Bu1 antibodies conjugated to PE and FITC, respectively. Analysis was performed in the gate according to forward/side scatter parameters as shown followed by KUL01-PE/Bu1-FICT. The results obtained in lamina propria analysis are shown, expressed as the mean (SD) in percentage and MFI (SD) from KUL01⁺ population from each group.