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. 2021 Jan 21;11:1864. doi: 10.1038/s41598-021-81389-z

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Identification of Tfh cells in lymph node cells from influenza infected ferrets. (a) Gating strategy to identify CD4 T cells in the ferret LN. Lymphocytes were identified by forward scatter area (FSC-A) and side-scatter area (SSC-A). Doublets were excluded by gating on single cells as determined by FSC-A versus FSC-H, live cells were identified by viability dye exclusion, CD4 T cells were identified as CD4+ CD79a−. For each step, the parental population is indicated above the plot. (b) Representative plots of ferret CD4 T cells stained with anti-BCL 6 (clone K112-91), anti-ferret CXCR5 (clone feX5-C05) and anti-ferret PD-1 (clone fePD-CL1) antibodies. The BCL6 expression of CXCR5++ PD-1++ CD4 T cells (Tfh, blue oval) were compared with CXCR5− PD-1− CD4 T cells (non-Tfh, red oval) cells. The parental population is indicated above the plot. (c) Representative plots of CD4 T cells from influenza-infected ferret, mouse and macaque LNs stained with species-appropriate CXCR5 and PD-1 antibodies (mouse, clones L138D7 and 29F.1A12 respectively; macaque, MU5UBEE and EH12.2H7 respectively). The parental population is indicated above the plot. (d) Representative plots of live lymphocytes from influenza-infected ferret, mouse and macaque LNs stained with their respective CXCR5 antibodies as well as CD79, B220 and CD19 antibodies. The parental population is indicated above the plot.