Abstract
Immunotoxicity has been an important topic in toxicology since inadvertent exposures to xenobiotics were found to alter immune functions in humans. While rodent toxicity tests can reveal some levels of immunotoxicity, alternative methods must be developed to identify the detailed mechanisms. In this study, a method of in vitro prediction of innate immune suppression by substances was developed using a genomics approach. The primary selection of immune suppressors was based on their ability to downregulate MCP-1, CCL3, TNF, IL-8, and IL-12p40 expression levels in lipopolysaccharide (LPS)-stimulated THP-1 cells. Among 11 substances classified as potent immune suppressors, six including dexamethasone, tacrolimus, tofacitinib, prednisolone, sodium lauryl sulfate, and benzoic acid were used to create a dataset by transcriptomics of chemical-treated THP-1 cells using bulk RNA sequencing. We selected genes that were significantly upregulated by suppressor treatment while filtering out genes also upregulated in LPS-treated THP-1 cells. We identified a 226-gene immunosuppressive gene set (ISG). Innate immune suppressor signature scores were calculated as the median expression of the ISG. In a validation dataset, the signature score predicted acyclovir, cyclosporine, and mercuric chloride as immune suppressors, while selecting genistein as a non-immune suppressor. Although more dataset integration is needed in the future, our results demonstrated the possibility and utility of a novel genomics-based approach, the transcriptome-based determination of innate immune suppressor (TDIS) assay, to evaluate innate immune suppression by different substances. This provides insight into the development of future alternative testing methods because it reflects a comprehensive genetic signature derived from multiple substances rather than one cytokine.
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Funding
This study was supported by grants (20183MFDS524) from the Ministry of Food and Drug Safety, Korea. S.S was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2020R1A4A2002903, NRF-2021R1A2C2010219, NRF-2017R1D1A1B04031161) and the SNUH Research Fund (0320210190).
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YN, YH, and SS conceptualized the study; YN, HQ, YH, and SS designed the experiments; YN and HQ performed the experiments; YN, HQ, and HJ analyzed the data; YN, HQ, HJ, KK, SL, and YH interpreted the data; YH provided reagents; and HJ contributed to manuscript writing. YN and HQ wrote the manuscript.
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Quan, H., Jun, H., Kim, K. et al. Development of a transcriptome-based determination of innate immune suppressor (TDIS) assay as an in vitro test for immunotoxicity. Arch Toxicol 97, 495–507 (2023). https://doi.org/10.1007/s00204-022-03406-y
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DOI: https://doi.org/10.1007/s00204-022-03406-y