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. 2022 Jun 20;13:889991. doi: 10.3389/fimmu.2022.889991

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Copyright © 2022 He, Chen, Hao, Yang and Shang

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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Recombinant chIL-9 induces proliferation of chicken CD3⁺ T cells. Chicken PBMCs (n = 3) were labelled with CFSE and then cultured for 5 days in the presence of the supernatant from mock transfected DF-1 cells (Control), pEGFP-chIL-9-transfected DF-1 cells (eukaryotic rchIL-9) at 1:1 dilution, purified rchIL-9 (100 ng/mL), and rchIL-2 (100 ng/mL), respectively. T cell proliferation was analyzed by flow cytometry. (A) Gating strategies to identify chicken T cells. Lymphocytes, single cells, live cells, and CD3⁺ or CD8α⁺ T-cell subsets were gated. (B) Representative dot-plots of proliferative total CD3⁺ T cells (upper panel), CD8α⁺CD3⁺ T cells and CD8α^-CD3⁺ T cells (lower panel) were shown based on the reduction in CFSE fluorescence (CFSE^low) between different groups. (C) The percentages of each proliferative T cells are compared between groups. Data are shown as mean ± SD. *p < 0.05, **p < 0.01, or ***p < 0.001.