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. 2020 Mar 2;10:3810. doi: 10.1038/s41598-020-60761-5

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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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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EV-A71 infection in primary human peripheral blood mononuclear cells (PBMCs). Human primary PBMCs (n = 5–9) were infected with mild, moderate, severe and heat-inactivated EV-A71 isolates at MOI 5, and harvested at 0, 6, 12 and 24 hpi for viral load quantification and flow cytometry. (a) Viral load levels were determined by qRT-PCR. (b,c) Percentage of EV-A71 Ag+ cells from total CD45+, CD3+ T cells, CD19+ B cells and CD14+ monocytes were determined via flow cytometry. (b) Bar graphs showing the levels of EV-A71 Ag+ cells from the different cell subsets. Data are presented as mean ± SEM. (c) Illustration of representative contour plots from one donor at 12 hpi. Statistical analysis was carried out with Kruskal-Wallis with Dunn’s multiple comparisons test to compare among EV-A71 isolates at the respective time-points (*p < 0.05; **p < 0.01; ***p < 0.001).