Abstract
Allergic asthma is promoted by type 2 inflammation involving cytokines such as IL-4, IL-5, and IL-13, with group 2 innate lymphoid cells (ILC2s) playing a key pathogenic role. Here, we identify T cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) as a negative regulator of ILC2 function. Tim-3 expression is upregulated in activated pulmonary ILC2s, and engagement with Tim-3 agonists inhibits ILC2 activation, proliferation, and type 2 cytokine production via the Nemo Like Kinase (NLK) signaling pathway and suppression of mitochondrial metabolism. In vivo, Tim-3 agonists alleviate airway hyperreactivity (AHR) and inflammation in both IL-33- and Alternaria alternata-induced AHR models, while ILC2-specific Tim-3 deletion exacerbates AHR. These results are confirmed in human ILC2s and humanized mice, supporting the translational relevance. Our findings establish Tim-3 as an inhibitory checkpoint for ILC2s and suggest its potential as a therapeutic target in allergic asthma and other ILC2-mediated diseases.
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Data availability
The bulk RNA-seq data generated in this study have been deposited in the Gene Expression Omnibus (GEO) database under accession number GSE298909. Additional information required to support the reanalysis of the data presented in this paper will be provided by the corresponding author upon reasonable request. Source data are provided with this paper.
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Acknowledgements
The authors thank Meng Li of the USC Libraries Bioinformatics Service for helping with data analysis. Bioinformatics software and computational resources used in the analysis were funded by the USC Office of Research and Norris Medical Library. This study was financially supported by R01 HL144790, R01 HL151493, R01 AI169687, R01 AI145813, R01 HL151769, and R01 HL159804 from the National Institutes of Health, Public Health Service (O.A.).
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Y.S. designed, performed, and analyzed the experiments, and wrote the first draft of the manuscript. Y.S., K.S., K.K., S.S., B.P.H., J.C. and G.Y. helped perform experiments and maintain animal husbandry for the experiments. X.L. performed RNA-seq analysis and pathway analysis. V.K. provided Havcr2flox/flox mice (Tim-3flox/flox) and Tim-3 agonist antibody. O.A. conceptualized, supervised, designed the experiments, interpreted the data, and finalized the manuscript. All authors critically read the manuscript.
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Sakano, Y., Sakano, K., Kokubo, K. et al. Tim-3 agonist restrains ILC2 function and attenuates airway hyperreactivity via NLK pathway. Nat Commun (2026). https://doi.org/10.1038/s41467-026-71336-9
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DOI: https://doi.org/10.1038/s41467-026-71336-9
