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. 2023 Mar 2;20(4):351–364. doi: 10.1038/s41423-023-00985-3

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© The Author(s) 2023

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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Fig. 4 — Felodipine, fasudil, imatinib, and caspofungin, screened by the SARS-CoV-2-S CAR-T-cell model, suppress cytokine release. A The flowchart for drug screening that can suppress SARS-CoV-2-induced inflammation based on the SARS-CoV-2-S CAR-T-cell model is shown. B S-293T, SARS-CoV-2-S CAR-T, and THP1 cells (1:10:10) were coincubated and treated with or without individual drugs (10 μM) from an FDA-approved library (~1049) before measuring the secretion of IL8 in the culture supernatants by ELISA. C The culture supernatants from drugs with more than 70% inhibition of IL8 secretion screened from B were further subjected to measurement of IFNγ by ELISA. D T cells were treated with drugs with more than 70% inhibition of IFNγ secretion screened from C for three days followed by toxicity assay (mean ± s.e.m). E S-293T, SARS-CoV-2-S CAR-T, and THP1 cells (1:10:10) were coincubated and treated with or without felodipine (#1), fasudil (#2), imatinib (#3), or caspofungin (#4) (10 μM) for three days before measuring the secretion of cytokines by a multiplex bead array. The inhibition rate is presented by a heatmap. F SARS-CoV-2-S CAR-T cells coincubated with S-293T cells (5:1) were treated with or without felodipine (#1), fasudil (#2), imatinib (#3), or caspofungin (#4) (10 μM) for the indicated duration, followed by a cytotoxicity assay (mean ± s.e.m). G SARS-CoV-2-S CAR-T cells were incubated with control 293 T or S-293T cells at a ratio of 3:1 for two days, and cells in suspension were subjected to RNA-seq analysis. Genes up- and downregulated by S-293T cells are shown in a pie chart (FDR < 0.01, FC > 1.5). H SARS-CoV-2-S CAR-T cells incubated with S-293T cells were treated with or without felodipine, fasudil, imatinib, or caspofungin (10 μM) for two days, and cells in suspension were subjected to RNA-seq analysis. Genes that were upregulated by S-293T cells but suppressed by felodipine, fasudil, imatinib, and caspofungin are shown. I Hallmark gene set enrichment analysis for overlapping genes (n = 865) as described in H. J, K SARS-CoV-2-S CAR-T cells were incubated with S-293T cells and treated with or without felodipine, fasudil, imatinib, or caspofungin (10 μM) for two days. T cells in suspension were subjected to RT‒qPCR analysis (J) to examine the expression of genes as indicated or immunoblotting analysis (K) using antibodies as indicated (Student’s t test, unpaired, two-tailed, *P < 0.05; **P < 0.01; ***P < 0.001)