key: cord-0946845-49rsoncw
authors: Mitsui, Akira; Sakai, Ryosei; Miwa, Kiyoshi; Shibahara, Susumu; Kurihara, Shigekazu; Nagao, Kenji
title: Elucidation of the antiviral mechanism of cystine and theanine through transcriptome analysis of mice and comparison with COVID-19 gene set data
date: 2020-07-09
journal: bioRxiv
DOI: 10.1101/2020.06.25.149427
sha: cbb2d62e148f5747c7fc7da256ab5fe98c0f2158
doc_id: 946845
cord_uid: 49rsoncw

We previously showed that oral administration of cystine and theanine (CT) to mice confers resistance to influenza virus infection. In human studies, CT prevented colds in healthy subjects and enhanced antibody production after influenza vaccination in elderly individuals with a poor nutritional status. The mechanism of action of CT is thought to be glutathione (GSH)-mediated regulation of intracellular redox, which might affect innate immune systems such as macrophages to exert physiological effects. The effect of CT on influenza is independent of viral type, and this treatment has a broad range of antiviral activities. To explore the mechanisms of CT in viral infection, we performed transcriptome profiling of spleen tissues isolated from influenza A virus (IAV)-infected mice. We identified unique gene signatures in response to CT in the IAV-infected mice. Genes upregulated by CT included redox-regulated genes such as GCLC/GCLM (subunits of glutamate cysteine ligase, a rate-limiting enzyme of GSH biosynthesis), TXN1, TXN2, TXNRD2, and SOD1, suggesting that the intracellular redox environment is substantially altered by CT. However, genes downregulated in response to CT included chemokine/chemokine receptor genes (CCL5, CCL19, CXCL9, CXCL12, CXCR3, CXCR4, and ACKR3), some of which are related to cytokine storm. A comparison with public COVID-19-related gene set data showed that the upregulated gene signature was highly similar to the downregulated gene sets of SARS-CoV/SARS-CoV-2-infected cells and the upregulated gene set of attenuated SARS-CoV-infected cells. In conclusion, the unique gene signatures observed in response to orally administered CT in IAV-infected mouse spleen tissues suggested that CT may attenuate viral infection, replication and associated symptoms such as cytokine storm.

used to determine the relationships with the gene sets of SARS-CoV/SARS-CoV-2-infected cells. Among 420 gene sets (as of June, 2020), the gene sets for GSE147507 [9] and GSE148729 and GSE30589 [15] [NCBI`s Gene Expression Omnibus (GEO) Series IDs] (https://www.ncbi.nlm.nih.gov/geo/) were selected. infection/replication, the response observed in IAV-CT_down also suggests that CT could attenuate viral infection and/or replication.

In this study, genes encoding chemokines, some of which are characteristic of cytokine storm, were downregulated in response to CT. Among these molecules, CCL5 and CXCL9 have been shown to be elevated in response to SARS-CoV-2, SARS-CoV or MERS-CoV infections[9] [21] . The IAV-CT_down signature also contained CXCL12 and CXCR4, which are thought to be involved in the activation of Th17 cells [22] [23] .

Our study has some limitations. For instance, we compared IAV and IAV-CT samples and therefore did not observe transcriptome changes in response to CT alone. Although we showed the downregulation of chemokine genes and virus-responsive genes, these changes could be due to the reduction of IAV infection and/or replication. Further work is needed to clarify the detailed mechanisms of the antiviral effects of CT.

In conclusion, the present study demonstrated the unique gene signatures in response to orally administered CT in IAV-infected mouse spleen tissues. Upregulated signatures included redox-related genes, and downregulated genes included chemokines. Based on a comparison with COVID-19-related public gene set data, CT may attenuate viral infection, replication and the cytokine storm related to COVID-19. Future clinical work is needed to confirm these findings. IAV-CON data (see Fig. 1B ) is overlaid with redox-related genes (red circles), chemokine/chemokine receptor genes (blue circles), HMC class I/II genes (purple squares), ISGs (green triangles) and HSP genes (red open triangles).

Combined administration of L-cystine and L-theanine enhances immune functions and protects against influenza virus infection in aged mice

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