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. 2021 Feb 17;11:4030. doi: 10.1038/s41598-021-83612-3

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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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Exhausted CD4⁺ T cell phenotype. Representative (n = 6) flow cytometry plots (A) demonstrating expression of cell surface markers (LAG-3, PD-1, TIGIT, TIM-3) indicating an exhaustive cell phenotype on CD4⁺ T cells characterised at day 0 (unstimulated, upper panels) and after 14 days stimulated with PHA (5.0 μg/ml, lower panels). Impact of PHA (5.0 μg/ml)-induced exhaustion upon expression of cell surface markers (PD-1, TIM-3, LAG-3, TIGIT, CTLA-4) on CD4⁺ T cells. Data presented as percentage of expressing CD4⁺ T cells (B), and MFI of CD4⁺ T cells (C), (mean + SEM, n = 4–10). The impact of PHA treatment on surface marker expression was significant (+ 2-tailed T-test, # Mann–Whitney U Test).