Synergistic induction of IL-23 by TNFα, IL-17A, and EGF in keratinocytes

doi:10.1016/j.cyto.2020.155357

Cytokine

Volume 138, February 2021, 155357

https://doi.org/10.1016/j.cyto.2020.155357 Get rights and content

Highlights

•
IL-23 is induced by TNFα and IL-17A in keratinocytes.
•
IL-23 induction by TNFα and IL-17A is enhanced in psoriatic keratinocytes.
•
EGFR signaling is required for effective IL-23 induction by TNFα and IL-17A in keratinocytes.
•
IL-23 induction by TNFα and IL-17A is specific to keratinocytes but not monocytes.

Abstract

IL-23 is an inflammatory cytokine that plays an essential role in Th17 immunity by enhancing Th17 cell proliferation and survival, and Th17 cytokine production. IL-23 has pathogenic roles in the development of Th17-mediated inflammatory diseases including psoriasis. Despite successful treatment of psoriasis by blocking IL-23, the regulation of IL-23 expression in psoriasis patients is largely unknown. Dendritic cells are generally considered to be the primary source of IL-23 in psoriasis. While high levels of IL-23 are found in psoriatic epidermis, IL-23 expression in psoriatic keratinoctyes remains a controversial issue. In this study, we demonstrated that IL-23 production is induced by a combination of TNFα and IL-17A in human keratinocytes. Additionally, this IL-23 induction by TNFα and IL-17A is further increased in psoriatic keratinocytes and is enhanced by EGFR signaling. Although IL-23 is also robustly induced by toll-like receptor agonists in dendritic cells and macrophages, IL-23 expression in these cell types is not regulated by TNFα, IL-17A, and EGFR signaling. Given that IL-23 is essential for maintaining Th17 activation, IL-23 induction by TNFα, IL-17A, and EGF in keratinocytes could play an important pathological role in psoriasis pathogenesis as well as the cutaneous rash associated with EGFR inhibition therapy.

Introduction

IL-23 is a heterodimeric cytokine composed of a unique p19 subunit and a p40 subunit that is shared with IL-12 [1]. It is an inflammatory cytokine essential for maintaining Th17 cell activity by enhancing Th17 cell proliferation, survival, and Th17 cytokine production [2]. IL-23 plays many critical roles in psoriasis pathogenesis, as demonstrated by its overexpression in psoriatic skin [3], the induction of psoriasis-like phenotypes in mice by intradermal IL-23 injection [4], the association of genetic variants in genes of IL-23 signaling with susceptibility or resistance to psoriasis [5], [6], and the alleviation of psoriatic conditions by anti–IL-23 antibodies in mouse models [7]. More importantly, antibodies against either IL23p40 or IL23p19 have demonstrated clinical efficacy in human psoriasis therapy [8], [9].

Despite the importance of IL-23 in psoriasis pathogenesis, the regulation of IL-23 expression in psoriasis remains largely elusive. IL-23 is primarily expressed in macrophages and dendritic cells in response to bacterial and viral infections and pathogen-associated molecular patterns [10], [11], [12], and dendritic cells are generally considered to be the primary source of IL-23 in psoriasis [3]. Despite high levels of IL-23 found in the epidermis of psoriatic skin [13], [14], [15], [16], [17], IL-23 expression in keratinoctyes remains a controversial issue. The expression of IL-23 mRNA has been detected in keratinocytes under different conditions [14], [18], [19]. However, only low levels of IL-23 protein were detected in keratinocytes under culture conditions that are not relevant to psoriasis [19]. To date, whether or not IL-23 protein is produced from keratinocytes remains a debatable issue. In this study, we aimed to determine whether IL-23 cytokine is expressed in keratinocytes and to identify the signals that trigger IL-23 expression in keratinocytes in an attempt to define the role of keratinocyte IL-23 expression in psoriasis pathogenesis.

Section snippets

Primary keratinocyte culture and reagents

Skin biopsy specimens (6 mm) were obtained from both the lesional skin of patients with psoriasis and the skin of individuals without psoriasis under an Oregon Health & Science University Institutional Review Board-approved protocol. The epidermis was trimmed off and minced into 1x1 mm² skin islets. The skin islets were planted on collagen-coated plates as described [20]. Keratinocytes from the outgrowth were trypsinized and plated in 12-well plates for the assay. The keratinocyte cultures were

IL-23 is induced by TNFα and IL-17A in human keratinocytes

To investigate the possibility of keratinocytes acting as a source of IL-23 production in psoriasis, we first analyzed the production of IL-23 by primary human keratinocytes in response to potential psoriatic cytokines including TNFα, IL-17A, IL-22, IL-17F, and interferonγ (IFN-γ). We found that maximal IL23p19 mRNA was induced by combined treatment with TNFα and IL-17A (Fig. 1A). Marginal IL23p19 expression was induced by either TNFα or IL-17A alone. IL23p19 expression was moderately induced

CRediT authorship contribution statement

Benjamin Ehst: Conceptualization, Resources, Writing - review & editing. Zhiping Wang: Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization. Justin Leitenberger: Resources. Danielle McClanahan: Resources. Rachel De La Torre: Data curation, Investigation. Erika Sawka: Resources. Alex G. Ortega-Loayza: Resources. Jennifer Strunck: Resources. Teri Greiling: Resources, Writing - review & editing. Eric Simpson: Resources, Writing - review & editing. Yuangang Liu:

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

We thank Melanie Swinson for procuring clinical samples and Clara Stemwedel for editing the manuscript. This study was partially supported by NIAMS R03 AR066736, and R01 AR070645.

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¹: Current address: Oregon Medical Research Center, Portland, OR 97223, USA.

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Synergistic induction of IL-23 by TNFα, IL-17A, and EGF in keratinocytes

Highlights

Abstract

Introduction

Section snippets

Primary keratinocyte culture and reagents

IL-23 is induced by TNFα and IL-17A in human keratinocytes

CRediT authorship contribution statement

Declaration of Competing Interest

Acknowledgments

Immunity

J. Invest. Dermatol.

J. Invest. Dermatol.

J. Invest. Dermatol.

Trends Immunol.

Cytokine

J. Invest. Dermatol.

J. Invest. Dermatol.

J. Invest. Dermatol.

Discovery of the IL-23/IL-17 signaling pathway and the treatment of psoriasis

J. Immunol.

Increased expression of interleukin 23 p19 and p40 in lesional skin of patients with psoriasis vulgaris

J. Exp. Med.

IL-23 stimulates epidermal hyperplasia via TNF and IL-20R2-dependent mechanisms with implications for psoriasis pathogenesis

J. Exp. Med.

Collaborative Association Study of Psoriasis, Genome-wide scan reveals association of psoriasis with IL-23 and NF-kappaB pathways

Nat. Genet.

Cutting edge: a critical functional role for IL-23 in psoriasis

J. Immunol.

CNTO 1275 Psoriasis Study Group, A human interleukin-12/23 monoclonal antibody for the treatment of psoriasis

N. Engl. J. Med.

A phase 2 trial of guselkumab versus adalimumab for plaque psoriasis

N. Engl. J. Med.

Regulation of virus-induced IL-12 and IL-23 expression in human macrophages

J. Immunol.

Commensal Gram-negative bacteria prime human dendritic cells for enhanced IL-23 and IL-27 expression and enhanced Th1 development

Eur. J. Immunol.

Differential regulation of interleukin 12 and interleukin 23 production in human dendritic cells

J. Exp. Med.

Low expression of the IL-23/Th17 pathway in atopic dermatitis compared to psoriasis

J. Immunol.

Epigenetic control of IL-23 expression in keratinocytes is important for chronic skin inflammation

Nat. Commun.