The secondary immunization was performed 21 days after the initial immunization. (c, d) The DBA mice were injected with AAV-control, AAV-shAhR or AAV-shCPT1 via the tail vein, and then were intradermally injected with type II collagen emulsion to establish collagen-induced arthritis (CIA) model. (b) The mRNA expression of IL-10 and Foxp3 in colon tissues was detected by using Q-PCR. DIM (50 mg/kg) was orally administered for consecutive 10 days. (a, b) The mice were injected with AAV-control, AAV-shAhR and AAV-shCPT1 via the tail vein for 14 days, and then were fed with 2.5% DSS for 7 days and then given normal drinking water for another 3 days. Involvement of FAO in the amelioration of weight loss and Treg generation by AhR activation in the mice with colitis and arthritis. The mRNA expression of Skp2 was detected by using Q-PCR. (a–d) The mRNA expression was detected by using Q-PCR. The naïve CD4+ T cells were transfected with negative control (NC) or siRNA for 24 h. Validation of siRNA knockdown efficiency. In summary, these findings indicate that AhR activation promotes Treg cell generation by enhancing Lkb1-mediated FAO via the Skp2/K63-ubiquitination pathway, and AhR agonists may be used as inducers of Treg cells to prevent and treat autoimmune diseases.įIGURE S1. Furthermore, the contribution of Skp2/K63 ubiquitination/Lkb1/FAO axis was verified in (DSS)-induced colitis or CIA mice. DIM and ITE were also shown to upregulate the mRNA expression of Skp2, a ubiquitination-related enzyme, and facilitate the binding of AhR to the xenobiotic responsive element of Skp2 promoter region by luciferase reporter gene assay. DIM and ITE functioned mainly through activating the liver kinase B1 (Lkb1)-AMPK pathway via promotion of Lkb1-Strad-Mo25 complex formation and Lkb1 K63 ubiquitination. Consistently, pharmacological or genetic inhibition of FAO markedly diminished the induction of DIM and ITE on the differentiation of Treg cells. Compared to the control group, exogenous and endogenous AhR agonists 3,3′-diindolylmethane (DIM) and 2-(1 ′H-indole-3′-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE) were shown to preferentially upregulate the mRNA expression of FAO-related enzymes and the value of OCR. AhR or carnitine palmitoyltransferases 1 was knockdown in dextran sulphate sodium (DSS)-induced colitis or collagen-induced arthritis (CIA) mice by infecting mice with adeno-associated virus via the tail vein injection. The binding of AhR to the Skp2 promoter was detected by constructing luciferase reporter gene. The Strad-Mo25-Lkb1 complex formation and K63 chain ubiquitination modification of Lkb1 were detected by co-immunoprecipitation. Intracellular localization of Lkb1 was detected by immunofluorescence. The activation of fatty acid oxidation (FAO)-related metabolic pathways was detected by Western blotting. The value of oxygen consumption rate (OCR) of CD4+ T cells was detected by the Seahorse XFe 96 analyzer. The proportions of Treg cells were detected by flow cytometry. Naïve CD4+ T cells were purified with mouse CD4+ CD62L+ T Cells Isolation Kits. In this study, we addressed the underlying mechanism of AhR activation to induce the generation of Treg cells in the view of cellular metabolism. Saving Earth Britannica Presents Earth’s To-Do List for the 21st Century.Several aryl hydrocarbon receptor (AhR) agonists have been reported to promote the generation of regulatory T cells (Treg cells), and the action mechanisms need to be identified.100 Women Britannica celebrates the centennial of the Nineteenth Amendment, highlighting suffragists and history-making politicians.COVID-19 Portal While this global health crisis continues to evolve, it can be useful to look to past pandemics to better understand how to respond today.Student Portal Britannica is the ultimate student resource for key school subjects like history, government, literature, and more.This Time in History In these videos, find out what happened this month (or any month!) in history.#WTFact Videos In #WTFact Britannica shares some of the most bizarre facts we can find.Demystified Videos In Demystified, Britannica has all the answers to your burning questions.Britannica Classics Check out these retro videos from Encyclopedia Britannica’s archives.Britannica Explains In these videos, Britannica explains a variety of topics and answers frequently asked questions.
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