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. 2020 Nov 23;10:20363. doi: 10.1038/s41598-020-76702-1

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© The Author(s) 2020

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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T cells from cancer patients exhibit lower CD4 + /CD8 + T cell ratio both in peripheral blood and after expansion. T cells purified from PBMCs of healthy individuals and cancer patients were analyzed for the surface expression of CD45RO, CD45RA, CD62L, CD28, CCR7, and CD127 using flow cytometry. IgG isotype control was used to assess non-specific binding. One of 12 representative experiments is shown in the figure (A). PBMCs isolated from the peripheral blood of healthy individuals and cancer patients were used to determine the surface expression of CD4 and CD8 using flow cytometry, and the percentages of CD4 + and CD8 + T cells were determined within CD3 + populations (n = 12) (B). The ratio of CD4:CD8 is shown in the figure (n = 12) (C). NK and T cells of healthy individuals and cancer patients were treated and co-cultured as described in Fig. 1A and Fig. 1E respectively. On days 6, 9, 12, and 15 of co-culture, the surface expression of CD4 and CD8 were analyzed by flow cytometry to obtain the percentages of CD4 + and CD8 + T cells within CD3 + populations (n = 28) (D). The ratio of CD4:CD8 is shown in the figure (n = 28) (E).