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Blockage of sphingosine-1-phosphate receptor 2 attenuates 2,4-dinitrochlorobenzene-induced atopic dermatitis in mice

Abstract

Sphingosine-1-phosphate (S1P) and its receptors have been implicated in functions of Langerhans cells and atopic dermatitis. In this study, we investigated the roles of S1P receptor type 2 (S1P2) in a mouse model of atopic dermatitis, which was induced by topical application of 2,4-dinitrochlorobenzene (DNCB) on ventral skin on D0, followed by repeated DNCB challenge on both ears from D7 to D49. Wild-type mice with atopic dermatitis displayed severe inflammation and mast cell accumulation in ear tissues and elevated IgE levels in serum. Furthermore, the mice showed significantly increased sizes of draining lymph nodes, high levels of inflammatory cytokines (IL-4, IL-13, IL-17, and IFN-γ) in the ears and lymph nodes and high levels of chemokines CCL17 and CCL22 in ears. Administration of JTE-013, a selective antagonist of S1P2 (3 mg/kg, i.p, from D19 to D49) before DNCB challenge significantly suppressed DNCB-induced atopic responses in ears and lymph nodes. JTE-013 administration also significantly decreased the lymph nodes sizes, the levels of inflammatory cytokines (IL-4, IL-13, IL-17, and IFN-γ) in the ears and lymph nodes, and the levels of chemokines CCL17 and CCL22 in ears. Furthermore, the inflammatory responses of atopic dermatitis were greatly ameliorated in S1pr2 gene-deficient mice. As CCL17 and CCL22 are CCR4 ligands, acting as Th2-attracting chemokines, we investigated CCL17 and CCL22 expression in bone marrow-derived dendritic cells (BMDCs) from wild-type and S1pr2 gene-deficient mice. Addition of IL-4 (10 ng/mL) markedly increased the levels of CCL17 and CCL22, but IL-4-induced CCL17 and CCL22 expression was significantly blunted in BMDCs from S1pr2 gene-deficient mice. Furthermore, pretreatment with JTE-013 (1−30 μM) dose-dependently suppressed this induction in BMDCs from wild-type mice. Our results demonstrate that blockage of S1P2 ameliorates not only DNCB-induced atopic dermatitis symptoms but also Th2 cell-attracting capacity of dendritic cells, suggesting S1P2 as a potential therapeutic target for atopic dermatitis.

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Fig. 1: Experimental protocol for the induction of atopic dermatitis and effect of S1P2 deficiency on DNCB-induced atopic dermatitis on mouse ears.
Fig. 2: Suppressive effect of S1P2 deficiency on the mRNA expression of cytokines in the ear samples.
Fig. 3: Effect of S1P2 deficiency on DNCB-induced atopic dermatitis in the lymph nodes.
Fig. 4: Suppressive effect of S1P2 deficiency on the mRNA expression of cytokines in the lymph nodes.
Fig. 5: Experimental protocol for the S1P2 antagonist experiment and the effect of S1P2 antagonist treatment on DNCB-induced atopic dermatitis responses in the mouse ears.
Fig. 6: Effect of S1P2 antagonist treatment on DNCB-induced atopic dermatitis responses in the lymph nodes.
Fig. 7: Inhibitory effects of an S1P2 antagonist and S1P2 deficiency on the expression of CCL17 and CCL22 in the mouse ear samples and BMDCs.
Fig. 8: Inhibitory effects of an S1P2 antagonist and S1P2 deficiency on the expression of CCL17 and CCL22 in the BMDCs.
Fig. 9: Suppressive effects of an S1P2 antagonist and S1P2 deficiency on IL-4 signaling in the BMDCs.

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Acknowledgements

This research was supported by the Basic Science Research Program of the Korean National Research Foundation funded by the Korean Ministry of Education, Science and Technology (NRF-2019R1A2C1005523).

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SJP and DSI designed the experiments. SJP performed the experiments and analyzed the data. SJP and DSI wrote the manuscript.

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Correspondence to Dong-Soon Im.

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Park, SJ., Im, DS. Blockage of sphingosine-1-phosphate receptor 2 attenuates 2,4-dinitrochlorobenzene-induced atopic dermatitis in mice. Acta Pharmacol Sin 41, 1487–1496 (2020). https://doi.org/10.1038/s41401-020-0412-8

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Keywords

  • atopic dermatitis
  • sphingosine-1-phosphate
  • sphingosine-1-phosphate receptor type 2 (S1P2)
  • S1pr2 gene-deficient mice
  • bone marrow-derived dendritic cell
  • cytokines

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