Journal of Investigative Dermatology Home

Nuclear IL-33 Plays an Important Role in IL-31‒Mediated Downregulation of FLG, Keratin 1, and Keratin 10 by Regulating Signal Transducer and Activator of Transcription 3 Activation in Human Keratinocytes

      IL-33, a chromatin-associated multifunctional cytokine, is implicated in the pathogenesis of atopic dermatitis (AD), an inflammatory skin disorder characterized by skin barrier dysfunction. IL-33 accumulates in the nuclei of epidermal keratinocytes (KCs) in AD lesions. However, it is unclear whether nuclear IL-33 directly contributes to the pathogenesis of AD. IL-31, a pruritogenic cytokine primarily produced by T helper type 2 cells, is elevated in AD lesions and promotes AD development by suppressing KC differentiation and inducing itching. In this study, we investigated the involvement of nuclear IL-33 in IL-31‒mediated suppression of KC differentiation. In monolayer cultures and living skin equivalent, IL-31 increased the expression of full-length IL-33 and the phosphorylation of signal transducer and activator of transcription 3 (STAT3) in the nuclei of human KCs, which in turn downregulated the expression of differentiation markers. We found that IL-31 and IL-4/IL-13 use very similar mechanisms to inhibit KC differentiation: nuclear IL-33 combines with phosphorylated STAT3 and functions as a STAT3 transcription cofactor, promoting phosphorylated STAT3 binding to the FLG promoter to inhibit its transcription; moreover, the nuclear IL-33/phosphorylated STAT3 complex drives the downregulation of keratin 1 and keratin 10 by reducing the availability of the transcription factor RunX1. Therefore, nuclear IL-33 plays an important role in IL-31‒mediated differentiation suppression by regulating STAT3 activation in human KCs.

      Graphical abstract


      AD (atopic dermatitis), AxSTAT3F (adenovirus vector expressing dominant-negative signal transducer and activator of transcription 3), ChIP (chromatin immunoprecipitation), ERK (extracellular signal‒regulated kinase), NHEK (normal human epidermal keratinocyte), K (keratin), KC (keratinocyte), LSE (living skin equivalent), p-STAT3 (phosphorylated signal transducer and activator of transcription 3), siRNA (small interfering RNA), STAT (signal transducer and activator of transcription), Th2 (T helper 2)
      To read this article in full you will need to make a payment

      Purchase one-time access:

      PDF Download and 24 Hours Online Access
      Society Members (SID/ESDR), remember to log in for access.


      Subscribe to Journal of Investigative Dermatology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Ali S.
        • Mohs A.
        • Thomas M.
        • Klare J.
        • Ross R.
        • Schmitz M.L.
        • et al.
        The dual function cytokine IL-33 interacts with the transcription factor NF-κB to dampen NF-κB-stimulated gene transcription.
        J Immunol. 2011; 187: 1609-1616
        • Amano W.
        • Nakajima S.
        • Kunugi H.
        • Numata Y.
        • Kitoh A.
        • Egawa G.
        • et al.
        The Janus kinase inhibitor JTE-052 improves skin barrier function through suppressing signal transducer and activator of transcription 3 signaling.
        J Allergy Clin Immunol. 2015; 136: 667-677.e7
        • Baekkevold E.S.
        • Roussigné M.
        • Yamanaka T.
        • Johansen F.E.
        • Jahnsen F.L.
        • Amalric F.
        • et al.
        Molecular characterization of NF-HEV, a nuclear factor preferentially expressed in human high endothelial venules.
        Am J Pathol. 2003; 163: 69-79
        • Balato A.
        • Raimondo A.
        • Balato N.
        • Ayala F.
        • Lembo S.
        Interleukin-33: increasing role in dermatological conditions.
        Arch Dermatol Res. 2016; 308: 287-296
        • Braun T.
        • Woollard A.
        RUNX factors in development: lessons from invertebrate model systems.
        Blood Cells Mol Dis. 2009; 43: 43-48
        • Cabanillas B.
        • Novak N.
        Atopic dermatitis and filaggrin.
        Curr Opin Immunol. 2016; 42: 1-8
        • Cayrol C.
        • Girard J.P.
        Interleukin-33 (IL-33): a nuclear cytokine from the IL-1 family.
        Immunol Rev. 2018; 281: 154-168
        • Cornelissen C.
        • Lüscher-Firzlaff J.
        • Baron J.M.
        • Lüscher B.
        Signaling by IL-31 and functional consequences.
        Eur J Cell Biol. 2012; 91: 552-566
        • Cornelissen C.
        • Marquardt Y.
        • Czaja K.
        • Wenzel J.
        • Frank J.
        • Lüscher-Firzlaff J.
        • et al.
        IL-31 regulates differentiation and filaggrin expression in human organotypic skin models.
        J Allergy Clin Immunol. 2012; 129: 433.e1-433.e8
        • Dai X.
        • Okazaki H.
        • Hanakawa Y.
        • Murakami M.
        • Tohyama M.
        • Shirakata Y.
        • et al.
        Eccrine sweat contains IL-1α, IL-1β and IL-31 and activates epidermal keratinocytes as a danger signal.
        PLoS One. 2013; 8: e67666
        • Dai X.
        • Tohyama M.
        • Murakami M.
        • Shiraishi K.
        • Liu S.
        • Mori H.
        • et al.
        House dust mite allergens induce interleukin 33 (IL-33) synthesis and release from keratinocytes via ATP-mediated extracellular signaling.
        Biochim Biophys Acta Mol Basis Dis. 2020; 1866: 165719
        • Dai X.
        • Utsunomiya R.
        • Shiraishi K.
        • Mori H.
        • Murakami M.
        • et al.
        Nuclear IL-33 plays an important role in the suppression of filaggrin, loricrin, keratin 1, and keratin 10 by IL-4 and IL-13 in human keratinocytes.
        J Invest Dermatol. 2021; (accessed August 1, 2021)
        • Dajnoki Z.
        • Béke G.
        • Mócsai G.
        • Kapitány A.
        • Gáspár K.
        • Hajdu K.
        • et al.
        Immune-mediated skin inflammation is similar in severe atopic dermatitis patients with or without filaggrin mutation.
        Acta Derm Venereol. 2016; 96: 645-650
        • Hänel K.H.
        • Pfaff C.M.
        • Cornelissen C.
        • Amann P.M.
        • Marquardt Y.
        • Czaja K.
        • et al.
        Control of the physical and antimicrobial skin barrier by an IL-31–IL-1 signaling network.
        J Immunol. 2016; 196: 3233-3244
        • Imai Y.
        Interleukin-33 in atopic dermatitis.
        J Dermatol Sci. 2019; 96: 2-7
        • Kim J.H.
        • Bae H.C.
        • Ko N.Y.
        • Lee S.H.
        • Jeong S.H.
        • Lee H.
        • et al.
        Thymic stromal lymphopoietin downregulates filaggrin expression by signal transducer and activator of transcription 3 (STAT3) and extracellular signal-regulated kinase (ERK) phosphorylation in keratinocytes.
        J Allergy Clin Immunol. 2015; 136: 205-208.e9
        • Kim S.
        • Kim H.J.
        • Yang H.S.
        • Kim E.
        • Huh I.S.
        • Yang J.M.
        IL-31 serum protein and tissue mRNA levels in patients with atopic dermatitis.
        Ann Dermatol. 2011; 23: 468-473
        • Lee H.
        • Shin J.J.
        • Bae H.C.
        • Ryu W.I.
        • Son S.W.
        Toluene downregulates filaggrin expression via the extracellular signal-regulated kinase and signal transducer and activator of transcription-dependent pathways.
        J Allergy Clin Immunol. 2017; 139: 355-358.e5
        • Lock F.E.
        • Babaian A.
        • Zhang Y.
        • Gagnier L.
        • Kuah S.
        • Weberling A.
        • et al.
        A novel isoform of IL-33 revealed by screening for transposable element promoted genes in human colorectal cancer.
        PLoS One. 2017; 12e0180659
        • Luzina I.G.
        • Fishelevich R.
        • Hampton B.S.
        • Courneya J.P.
        • Parisella F.R.
        • Lugkey K.N.
        • et al.
        Full-length IL-33 regulates Smad3 phosphorylation and gene transcription in a distinctive AP2-dependent manner.
        Cell Immunol. 2020; 357: 104203
        • Luzina I.G.
        • Pickering E.M.
        • Kopach P.
        • Kang P.H.
        • Lockatell V.
        • Todd N.W.
        • et al.
        Full-length IL-33 promotes inflammation but not Th2 response in vivo in an ST2-independent fashion.
        J Immunol. 2012; 189: 403-410
        • Masse I.
        • Barbollat-Boutrand L.
        • Molina M.
        • Berthier-Vergnes O.
        • Joly-Tonetti N.
        • Martin M.T.
        • et al.
        Functional interplay between p63 and p53 controls RUNX1 function in the transition from proliferation to differentiation in human keratinocytes.
        Cell Death Dis. 2012; 3: e318
        • Mitamura Y.
        • Nunomura S.
        • Nanri Y.
        • Ogawa M.
        • Yoshihara T.
        • Masuoka M.
        • et al.
        The IL-13/periostin/IL-24 pathway causes epidermal barrier dysfunction in allergic skin inflammation.
        Allergy. 2018; 73: 1881-1891
        • Nakashima C.
        • Otsuka A.
        • Kabashima K.
        Interleukin-31 and interleukin-31 receptor: new therapeutic targets for atopic dermatitis.
        Exp Dermatol. 2018; 27: 327-331
        • Raap U.
        • Wichmann K.
        • Bruder M.
        • Ständer S.
        • Wedi B.
        • Kapp A.
        • et al.
        Correlation of IL-31 serum levels with severity of atopic dermatitis.
        J Allergy Clin Immunol. 2008; 122: 421-423
        • Ryu W.I.
        • Lee H.
        • Bae H.C.
        • Ryu H.J.
        • Son S.W.
        IL-33 down-regulates filaggrin expression by inducing STAT3 and ERK phosphorylation in human keratinocytes.
        J Dermatol Sci. 2016; 82: 131-134
        • Sa S.M.
        • Valdez P.A.
        • Wu J.
        • Jung K.
        • Zhong F.
        • Hall L.
        • et al.
        The effects of IL-20 subfamily cytokines on reconstituted human epidermis suggest potential roles in cutaneous innate defense and pathogenic adaptive immunity in psoriasis.
        J Immunol. 2007; 178: 2229-2240
        • Szalus K.
        • Trzeciak M.
        • Nowicki R.J.
        JAK-STAT inhibitors in atopic dermatitis from pathogenesis to clinical trials results.
        Microorganisms. 2020; 8: 1743
        • Tamagawa-Mineoka R.
        • Okuzawa Y.
        • Masuda K.
        • Katoh N.
        Increased serum levels of interleukin 33 in patients with atopic dermatitis.
        J Am Acad Dermatol. 2014; 70: 882-888

      Supplementary References

        • Dai X.
        • Sayama K.
        • Yamasaki K.
        • Tohyama M.
        • Shirakata Y.
        • Hanakawa Y.
        • et al.
        SOCS1-negative feedback of STAT1 activation is a key pathway in the dsRNA-induced innate immune response of human keratinocytes.
        J Invest Dermatol. 2006; 126: 1574-1581
        • Shirakata Y.
        • Ueno H.
        • Hanakawa Y.
        • Kameda K.
        • Yamasaki K.
        • Tokumaru S.
        • et al.
        TGF-beta is not involved in early phase growth inhibition of keratinocytes by 1alpha,25(OH)2vitamin D3.
        J Dermatol Sci. 2004; 36: 41-50
        • Tohyama M.
        • Yang L.
        • Hanakawa Y.
        • Dai X.
        • Shirakata Y.
        • Sayama K.
        IFN-alpha enhances IL-22 receptor expression in keratinocytes: a possible role in the development of psoriasis.
        J Invest Dermatol. 2012; 132: 1933-1935
        • Yamasaki K.
        • Hanakawa Y.
        • Tokumaru S.
        • Shirakata Y.
        • Sayama K.
        • Hanada T.
        • et al.
        Suppressor of cytokine signaling 1/JAB and suppressor of cytokine signaling 3/cytokine-inducible SH2 containing protein 3 negatively regulate the signal transducers and activators of transcription signaling pathway in normal human epidermal keratinocytes.
        J Invest Dermatol. 2003; 120: 571-580