Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

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


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.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

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.


  1. 1.

    Davidson WF, Leung DYM, Beck LA, Berin CM, Boguniewicz M, Busse WW, et al. Report from the National Institute of Allergy and Infectious Diseases workshop on “Atopic dermatitis and the atopic march: Mechanisms and interventions”. J Allergy Clin Immunol. 2019;143:894–913.

    PubMed Central  Google Scholar 

  2. 2.

    Leung DY, Guttman-Yassky E. Assessing the current treatment of atopic dermatitis: Unmet needs. J Allergy Clin Immunol. 2017;139:S47–8.

    Google Scholar 

  3. 3.

    Baumer W, Rossbach K, Mischke R, Reines I, Langbein-Detsch I, Luth A, et al. Decreased concentration and enhanced metabolism of sphingosine-1-phosphate in lesional skin of dogs with atopic dermatitis: disturbed sphingosine-1-phosphate homeostasis in atopic dermatitis. J Invest Dermatol. 2011;131:266–8.

    Google Scholar 

  4. 4.

    Park SJ, Im DS. Sphingosine 1-phosphate receptor modulators and drug discovery. Biomol Ther. 2017;25:80–90.

    CAS  Google Scholar 

  5. 5.

    Leong WI, Saba JD. S1P metabolism in cancer and other pathological conditions. Biochimie. 2010;92:716–23.

    CAS  PubMed Central  Google Scholar 

  6. 6.

    Spiegel S, Milstien S. Functions of the multifaceted family of sphingosine kinases and some close relatives. J Biol Chem. 2007;282:2125–9.

    CAS  Google Scholar 

  7. 7.

    Bock S, Pfalzgraff A, Weindl G. Sphingosine 1-phospate differentially modulates maturation and function of human Langerhans-like cells. J Dermatol Sci. 2016;82:9–17.

    CAS  Google Scholar 

  8. 8.

    Kohno T, Tsuji T, Hirayama K, Watabe K, Matsumoto A, Kohno T, et al. A novel immunomodulator, FTY720, prevents spontaneous dermatitis in NC/Nga mice. Biol Pharm Bull. 2004;27:1392–6.

    CAS  Google Scholar 

  9. 9.

    Yanagawa Y, Hoshino Y, Kataoka H, Kawaguchi T, Ohtsuki M, Sugahara K, et al. FTY720, a novel immunosuppressant, prolongs rat skin allograft survival by decreasing T-cell infiltration into grafts. Transpl Proc. 1999;31:1227–9.

    CAS  Google Scholar 

  10. 10.

    Checa A, Xu N, Sar DG, Haeggstrom JZ, Stahle M, Wheelock CE. Circulating levels of sphingosine-1-phosphate are elevated in severe, but not mild psoriasis and are unresponsive to anti-TNF-α treatment. Sci Rep. 2015;5:12017.

    CAS  Article  PubMed Central  Google Scholar 

  11. 11.

    Mysliwiec H, Baran A, Harasim-Symbor E, Choromanska B, Mysliwiec P, Milewska AJ, et al. Increase in circulating sphingosine-1-phosphate and decrease in ceramide levels in psoriatic patients. Arch Dermatol Res. 2017;309:79–86.

    CAS  Article  Google Scholar 

  12. 12.

    Schaper K, Dickhaut J, Japtok L, Kietzmann M, Mischke R, Kleuser B, et al. Sphingosine-1-phosphate exhibits anti-proliferative and anti-inflammatory effects in mouse models of psoriasis. J Dermatol Sci. 2013;71:29–36.

    CAS  Article  Google Scholar 

  13. 13.

    Liao JJ, Huang MC, Goetzl EJ. Cutting edge: alternative signaling of Th17 cell development by sphingosine 1-phosphate. J Immunol. 2007;178:5425–8.

    CAS  Article  Google Scholar 

  14. 14.

    Shin SH, Cho KA, Hahn S, Lee Y, Kim YH, Woo SY, et al. Inhibiting Sphingosine Kinase 2 Derived-sphingosine-1-phosphate ameliorates psoriasis-like skin disease via blocking Th17 differentiation of naive CD4 T lymphocytes in mice. Acta Derm Venereol. 2019;99:594–601.

    CAS  Article  Google Scholar 

  15. 15.

    Reines I, Kietzmann M, Mischke R, Tschernig T, Luth A, Kleuser B, et al. Topical application of sphingosine-1-phosphate and FTY720 attenuate allergic contact dermatitis reaction through inhibition of dendritic cell migration. J Invest Dermatol. 2009;129:1954–62.

    CAS  Article  Google Scholar 

  16. 16.

    Nakashima D, Kabashima K, Sakabe J, Sugita K, Kobayashi T, Yoshiki R, et al. Impaired initiation of contact hypersensitivity by FTY720. J Invest Dermatol. 2008;128:2833–41.

    CAS  Article  Google Scholar 

  17. 17.

    Schumann J, Grevot A, Ledieu D, Wolf A, Schubart A, Piaia A, et al. Reduced activity of sphingosine-1-phosphate lyase induces podocyte-related glomerular proteinuria, skin irritation, and platelet activation. Toxicol Pathol. 2015;43:694–703.

    CAS  Article  PubMed Central  Google Scholar 

  18. 18.

    Japtok L, Schaper K, Baumer W, Radeke HH, Jeong SK, Kleuser B. Sphingosine 1-phosphate modulates antigen capture by murine Langerhans cells via the S1P2 receptor subtype. PLoS ONE. 2012;7:e49427.

    CAS  Article  PubMed Central  Google Scholar 

  19. 19.

    Kono M, Belyantseva IA, Skoura A, Frolenkov GI, Starost MF, Dreier JL, et al. Deafness and stria vascularis defects in S1P2 receptor-null mice. J Biol Chem. 2007;282:10690–6.

    CAS  Article  PubMed Central  Google Scholar 

  20. 20.

    Park SJ, Im DS. Deficiency of sphingosine-1-phosphate receptor 2 (S1P2) attenuates bleomycin-induced pulmonary fibrosis. Biomol Ther. 2018;27:318–26.

  21. 21.

    Park SJ, Im DS. Blockage of sphingosine-1-phosphate receptor 2 attenuates allergic asthma in mice. Br J Pharmacol. 2019;176:938–49.

    CAS  PubMed Central  Google Scholar 

  22. 22.

    Lutz MB, Kukutsch N, Ogilvie AL, Rossner S, Koch F, Romani N, et al. An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods. 1999;223:77–92.

    CAS  Google Scholar 

  23. 23.

    Olivera A, Dillahunt SE, Rivera J. Interrogation of sphingosine-1-phosphate receptor 2 function in vivo reveals a prominent role in the recovery from IgE and IgG-mediated anaphylaxis with minimal effect on its onset. Immunol Lett. 2013;150:89–96.

    CAS  PubMed Central  Google Scholar 

  24. 24.

    Oskeritzian CA, Price MM, Hait NC, Kapitonov D, Falanga YT, Morales JK, et al. Essential roles of sphingosine-1-phosphate receptor 2 in human mast cell activation, anaphylaxis, and pulmonary edema. J Exp Med. 2010;207:465–74.

    CAS  PubMed Central  Google Scholar 

  25. 25.

    Kim JY, Jeong MS, Park MK, Lee MK, Seo SJ. Time-dependent progression from the acute to chronic phases in atopic dermatitis induced by epicutaneous allergen stimulation in NC/Nga mice. Exp Dermatol. 2014;23:53–7.

    Google Scholar 

  26. 26.

    Koga C, Kabashima K, Shiraishi N, Kobayashi M, Tokura Y. Possible pathogenic role of Th17 cells for atopic dermatitis. J Invest Dermatol. 2008;128:2625–30.

    CAS  Google Scholar 

  27. 27.

    Muraro A, Lemanske RF Jr., Hellings PW, Akdis CA, Bieber T, Casale TB, et al. Precision medicine in patients with allergic diseases: Airway diseases and atopic dermatitis-PRACTALL document of the European Academy of Allergy and Clinical Immunology and the American Academy of Allergy, Asthma & Immunology. J Allergy Clin Immunol. 2016;137:1347–58.

    Google Scholar 

  28. 28.

    Sawicka E, Zuany-Amorim C, Manlius C, Trifilieff A, Brinkmann V, Kemeny DM, et al. Inhibition of Th1- and Th2-mediated airway inflammation by the sphingosine 1-phosphate receptor agonist FTY720. J Immunol. 2003;171:6206–14.

    CAS  Google Scholar 

  29. 29.

    Matsuo K, Hatanaka S, Kimura Y, Hara Y, Nishiwaki K, Quan YS, et al. A CCR4 antagonist ameliorates atopic dermatitis-like skin lesions induced by dibutyl phthalate and a hydrogel patch containing ovalbumin. Biomed Pharmacother. 2019;109:1437–44.

    CAS  Google Scholar 

  30. 30.

    Zlotnik A, Yoshie O. Chemokines: a new classification system and their role in immunity. Immunity. 2000;12:121–7.

    CAS  Google Scholar 

  31. 31.

    Stutte S, Quast T, Gerbitzki N, Savinko T, Novak N, Reifenberger J, et al. Requirement of CCL17 for CCR7- and CXCR4-dependent migration of cutaneous dendritic cells. Proc Natl Acad Sci USA. 2010;107:8736–41.

    CAS  Google Scholar 

  32. 32.

    Homey B, Steinhoff M, Ruzicka T, Leung DY. Cytokines and chemokines orchestrate atopic skin inflammation. J Allergy Clin Immunol. 2006;118:178–89.

    CAS  Article  Google Scholar 

  33. 33.

    Tsuji T, Yoshida Y, Iwatsuki R, Inoue M, Fujita T, Kohno T. Therapeutic approach to steroid-resistant dermatitis using novel immunomodulator FTY720 (Fingolimod) in combination with betamethasone ointment in NC/Nga mice. Biol Pharm Bull. 2012;35:1314–9.

    CAS  Article  Google Scholar 

  34. 34.

    Tsuji T, Okuno S, Kuroda A, Hamazaki J, Chikami T, Sakurai S, et al. Therapeutic approach to mite-induced intractable dermatitis using novel immunomodulator FTY720 ointment (fingolimod) in NC/Nga mice. Allergol Int. 2016;65:172–9.

    CAS  Article  Google Scholar 

  35. 35.

    Kim DS, Kim SY, Kleuser B, Schafer-Korting M, Kim KH, Park KC. Sphingosine-1-phosphate inhibits human keratinocyte proliferation via Akt/protein kinase B inactivation. Cell Signal. 2004;16:89–95.

    CAS  Article  Google Scholar 

  36. 36.

    Jeong SK, Kim YI, Shin KO, Kim BW, Lee SH, Jeon JE, et al. Sphingosine kinase 1 activation enhances epidermal innate immunity through sphingosine-1-phosphate stimulation of cathelicidin production. J Dermatol Sci. 2015;79:229–34.

    CAS  Article  PubMed Central  Google Scholar 

  37. 37.

    Jeon S, Song J, Lee D, Kim GT, Park SH, Shin DY, et al. Inhibition of sphingosine 1-phosphate lyase activates human keratinocyte differentiation and attenuates psoriasis in mice. J Lipid Res. 2020;61:20–32.

    CAS  Article  Google Scholar 

Download references


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).

Author information




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

Corresponding author

Correspondence to Dong-Soon Im.

Ethics declarations

Competing interests

The authors declare no competing interests.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

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).

Download citation


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


Quick links