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. 2020 Aug 14;10:13843. doi: 10.1038/s41598-020-70673-z

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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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Differential gene expression in HSV-specific CD8⁺ T cells from HSV-1 infected symptomatic vs. asymptomatic individuals. (a) Experimental design and validation of differentially expressed genes in CD8⁺ T cells sharing the same HSV-1 epitope-specificities, from SYMP and ASYMP individuals. CD8⁺ T cells specific to HLA-A*0201-restricted HSV-1 gB_561–567 and VP11/12_702–710 epitopes were sorted from HLA-A*0201-positive SYMP and ASYMP individuals, using specific tetramers. Total RNA was extracted from each clone of epitope-specific CD8⁺ T cells, and whole transcriptome analysis was performed using bulk RNA sequencing to determine the levels of expression of 25,638 genes. (b) Frequencies of CD8⁺ T cells specific to HLA-A*0201-restricted HSV-1 gB_561–567 and VP11/12_702–710 epitopes detected by FACS in SYMP vs. ASYMP individuals. (c) Heatmap is showing 772 differentially expressed genes among SYMP and ASYMP individuals. (d) Heatmap showing statistically significant pathways that are affected in HSV-specific CD8⁺ T cells from SYMP vs. SYMP individuals. Parametric Gene Set Enrichment Analysis (PSGEA) method was applied based on data curated in Gene Ontology and KEGG. Pathway significance cut-off with a false discovery date (FDR) ≥ 0.2 was applied. (e) Bulk RNA heatmap comparing differentially expressed CAM pathway associated T cell co-stimulatory and T cell exhaustion genes in HSV-specific CD8⁺ T cells from SYMP vs. SYMP individuals.