- About this Journal ·
- Abstracting and Indexing ·
- Aims and Scope ·
- Annual Issues ·
- Article Processing Charges ·
- Articles in Press ·
- Author Guidelines ·
- Bibliographic Information ·
- Citations to this Journal ·
- Contact Information ·
- Editorial Board ·
- Editorial Workflow ·
- Free eTOC Alerts ·
- Publication Ethics ·
- Reviewers Acknowledgment ·
- Submit a Manuscript ·
- Subscription Information ·
- Table of Contents
Journal of Allergy
Volume 2012 (2012), Article ID 718725, 10 pages
Keratinocytes under Fire of Proinflammatory Cytokines: Bona Fide Innate Immune Cells Involved in the Physiopathology of Chronic Atopic Dermatitis and Psoriasis
1Laboratoire Inflammation, Tissus Épithéliaux et Cytokines, UPRES-EA 4331, CHU de Poitiers, Pole Biologie Santé, Université de Poitiers, Bâtiment B36, 1 rue G Bonnet, 86022 Poitiers, France
2BIOalternatives, 1 bis rue des Plantes, 86160 Gençay, France
Received 15 June 2012; Revised 5 October 2012; Accepted 5 October 2012
Academic Editor: Maria Leite-de-Moraes
Copyright © 2012 François-Xavier Bernard et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Cutaneous homeostasis and defenses are maintained by permanent cross-talk among particular epidermal keratinocytes and immune cells residing or recruited in the skin, through the production of cytokines. If required, a coordinated inflammatory response is triggered, relayed by specific cytokines. Due to numerous reasons, troubles in the resolution of this phenomenon could generate a cytokine-mediated vicious circle, promoting skin chronic inflammation, the most common being atopic dermatitis and psoriasis. In this paper, we discuss the biological effects of cytokine on keratinocytes, more particularly on specific or shared cytokines involved in atopic dermatitis or psoriasis. We report and discuss monolayer or 3D in vitro models of keratinocytes stimulated by specific sets of cytokines to mimic atopic dermatitis or psoriasis. IL-22, TNFa, IL-4, and IL-13 combination is able to mimic an “atopic dermatitis like” state. In psoriasis lesions, over expression of IL-17 is observed whereas IL-4 and IL-13 were not detected; the replacement of IL-4 and IL-13 by IL-17 from this mix is able to mimic in vitro a “psoriasis like” status on keratinocytes. We conclude that specific cytokine environment deregulation plays a central role on skin morphology and innate immunity, moving towards specific pathologies and opening the way to new therapeutic strategies.
Skin constitutes the largest physical and chemical barrier against various stresses including pathogens, constituting the first line of defense of the body. Cutaneous homeostasis and defenses are maintained by permanent cross-talk among dermal fibroblasts, epidermal keratinocytes, and cells of the immune system residing or recruited in the skin, through the production of cytokines. Recent years showed that skin appears as a multitasking immune organ, and highlighted the key role of keratinocytes, which are the epithelial cells of epidermis (the outer skin layer in contact with environment), in this protection . Keratinocytes are no longer considered as a passive protection barrier but as true innate immune cells. Indeed, skin represents a very attractive tissue that is a paradigm for studying the cross-talk between innate and adaptive immunity system and an organ.
To be effective for protection against a large panel of injuries, skin requires sensitivity and selectivity to detect a signal of danger, a strong reactivity to develop a rapid response, and efficiency. These requirements need sophisticated interactions between keratinocytes and “sentinel” immune cells infiltrating normal epidermis, that is, T lymphocytes and Langerhans cells . These interactions are conducted by cytokines, maintaining the homeostasis of skin; if required, a coordinated inflammatory response is triggered, relayed by specific cytokines. Due to a number of known or unknown reasons (genetic, environmental, etc.), troubles in the resolution of this phenomenon could generate a cytokine-mediated vicious circle, promoting chronic inflammation. This state is characterized by resident/infiltrating immune cells in the epidermis or dermis, altered differentiation of keratinocytes and the increase of epidermal thickness in some cases. They are the result in particular of an altered dialogue between keratinocytes and activated immune cells, due on a balance of Th subsets, especially Th1, Th2, Th17, and Th22 producing specific sets of cytokines. Various inflammatory responses can be observed, leading to different clinical entities, the most common being atopic dermatitis (AD) and psoriasis. AD affect up to 3% of adults and 25% of children, and psoriasis 2.5% of the world’s population . Both are classically considered as two opposing models due to the polarization of the Th response.
Atopic dermatitis has a complex pathogenesis associating inflammatory reactions to epidermal barrier dysfunctions, allowing allergen sensitization. Genetic studies showed the importance of filaggrin in AD . Filaggrin is a protein of the stratum corneum involved in the maintenance of keratin cytoskeleton, the assembly of the cornified envelope, and the water-binding capacities of skin. Inflammatory cytokines are implicated in skin barrier disruption by downregulating the protein expression of the cornified envelop, including filaggrin.
Chronic lesions of AD are characterized by lichenification with skin thickening and hyperplasia of the epidermis.
Plaques of psoriasis are characterized by parakeratosis, but also skin thickening and acanthotic epidermis with abundant dermal mononuclear cell infiltrate. These two inflammatory chronic skin disorders were opposed by the long standing Th1/Th2 paradigm, despite the fact that they share some histological similarities. The discovery of a role for IL-17-producing T helper cells (Th17) in psoriasis and for IL-22 producing (Th22) cells in AD emphasize the complexity of cytokine network involved in the induction and/or maintenance of these disorders.
Our paper is focused on the cytokines associated to AD compared to those in psoriasis and their direct biological effects on keratinocytes. For a general recent review comparing clinical features, immune cells, and therapeutic strategies in AD and psoriasis, refer to Guttman-Yassky et al. [2, 3]. Likewise, review and the experimental studies reported herein are focused on the chronic phases, occulting the description of initiating events and earlier phases of these skin diseases.
We study in vitro the respective effects of AD or psoriasis-associated cytokines on parameters of innate immunity and histology using the (normal human epidermal keratinocytes) NHEKs and (reconstituted human epidermis) RHE, and discussed the results in the context of Th polarization. We shall see that the inflammatory skin phenotype is largely the consequence of the effect of cytokines on keratinocytes. We will comment on the view that, if AD and psoriasis were opposed by long standing paradigms, they also have common features.
2. Cytokines and Keratinocytes
During the past few years, an increasing number of reports demonstrated that keratinocytes are direct targets for a specific set of cytokines, conducting dramatic changes in their biological properties such as inducing the secretion of chemokines and antimicrobial peptides or modulating the differentiation states or migration capacities. In the 1990s, functional receptors for the classical inflammatory cytokines interleukin (IL)-1 and tumor necrosis factor (TNF) or the Th1-derived interferon (IFN)-γ have been described on keratinocytes (review in ). More recently, IL-17, IL-21, IL-22, and oncostatin M (OSM) have also been described as important regulators of epidermis functions, and their receptors have been detected on keratinocytes [6–9]. Other cytokines are able to induce skin inflammation in animal models, as do IL-12 or IL-23. However, keratinocytes do not display functional receptors for IL-12 and IL-23 , suggesting that other cell types are the targets, and that the effect on keratinocytes is indirect.
As an example, amongst the cytokines of the IL-10 family, IL-22 is the strongest activator of keratinocytes, and to a lesser extent IL-24, followed by IL-20 and IL-19 . Analysis of the gene expression profile induced on NHEK showed that these cytokines upregulate the expression of genes associated with inflammation and innate immunity, such as S100A7-psoriasin and β-defensin2 (BD-2); they also downregulate differentiation-associated genes, including CK10, and induce hyperplasia and hypogranulosis of RHE [11, 12] and upregulated CK16, which is associated with suprabasal keratinocyte proliferation.
The simultaneous analysis of a large panel of cytokines demonstrate that a number of them directly target keratinocytes, that is, cytokines of the IL-1 (IL-1α and β), IL-2 (IL-4, IL-13, IL-21), IL-4 (IL-4, IL-13), IL-6 (IL-6, OSM, IL-31), IL-10 (IL-19, IL-20, IL-22, IL-24), IL-17 (IL-17A and IL-17F), IFN (IFNα, IFNγ), or TNF (TNFα, TNFβ) family . Note that the biological effect of IL-4 or IL-13 on the inhibition of defensin production is observed only after preactivation of keratinocytes by the IL-4, IL-13, IL-22, and TNFα cytokine combination, which enhance IL-13RA2 chain expression as defined below. Regarding IL-21 receptor, IFNα and γ enhance the IL-2Rγ chain expression; since IL-21R is also expressed by keratinocytes (unpublished data), it suggest the sensitivity of keratinocytes to IL-21. It has been confirmed by Caruso et al, demonstrating that IL-21 induces in vitro the proliferation of isolated keratinocytes  and induces Erk phosphorylation (unpublished data). In contrast and in our hands, there is no evidence for a direct effect IL-2, IL-7, IL-9, and IL-15 on keratinocytes . Transcriptomic analysis of the keratinocyte gene expression profile demonstrates that most cytokines active on keratinocytes have overlapping activities, sometimes displaying completely redundant biological properties. The use of shared molecules in intracellular signaling pathways, such as STAT3 in OSM or IL-22 signaling or NFκB in IL-1 or TNF signaling could explain redundancy [6, 7]. Recently, the IL-36 members of the IL-1 family have been reported to induce by an autocrine effect their own secretion and antibacterial peptides production by keratinocytes .
Usually upstream of cytokine action during an inflammatory process, TLR engagement activates NFκB pathways, as IL-1 or TNF receptors engagement do, and may have partial redundant activities with these cytokines on keratinocytes. Keratinocytes express TLR 1-6 and 9 [16, 17]. TLR engagement have been described to induce cytokines, such as TSLP  and IFNα and β. Synergistic and/or overlapping biological activities of TLR ligands and cytokines on keratinocytes are very likely, as discussed below for cytokine combinations. Poorly documented up to now, the study of these costimulations could give a more dynamic view of the course of skin inflammation.
In another hand, powerful synergistic effects on keratinocytes have been also evidenced when cytokines of different families are associated . These cytokines modified the expression of genes associated with inflammation, innate immunity, and differentiation. However, comparative and quantitative analysis demonstrates huge differences of expression induced by specific cytokines. For example, IL-22, as OSM, both via the STAT3 activation pathway induced a strong inhibition of keratinocyte differentiation, as objectived by the decreased expression of filaggrin, loricrin and involucrin and the increase in the overall thickness of RHE [6, 7]. In contrast, the induction of chemokines, S100A7, or BD-2 by IL-22 and OSM is moderated. On the other hand, whereas IL-17, IL-1, and TNFα strongly induced chemokines, S100A7, or BD-2 expression, they have weaker effect on keratinocyte differentiation [14, 19]. Nevertheless, it appears that single cytokine stimulation generates a rather limited effect on keratinocytes, namely, a limited number and/or a limited modulated expression of targeted genes.
Since in physiological or physiopathological conditions, tissues are surrounded not by one cytokine but a complex milieu, study of the biological activities of cytokine combinations is of great interest. For example, combination of IL-17A and IFN-γ or IL-17A and TNF-α results in a synergistic effect on CXCL8 production by keratinocytes [20, 21]. IL-17A and IL-22 synergize in the upregulation of BD-2 and S100A9 production [22, 23]. The association of IL-1α, IL-17, IL-22, OSM, and TNFα demonstrated a very strong synergy in increasing the expression of inflammatory molecules such as psoriasin/S100A7 or BD-2, or IL-8 in vitro by NHEK . When IL-22 is removed of the cytokine mixture, CXCL8 and BD-2 expression is reduced by 30%, whereas the decrease is about 70% after IL-17 removing. In addition, ex vivo studies on human skin explants demonstrated upregulation of BD-2, S100A7, and CXCL8 expression in response to the same combination of cytokines. In vivo intradermal injection of these five cytokines in mouse increased CXCL1, CXCL2, CXCL3, S100A9, and BD-3 expression, associated with neutrophil infiltration and an early epidermal acanthosis  (and submitted).
3. Specific and Shared Cytokines in Atopic Dermatitis and Psoriasis
The presence of CD4+CCR4+ Th2-cells in lesional skin of acute atopic dermatitis patients was described . Th2 secreted predominantly IL-4 and IL-13, which induce both pro- and anti-inflammatory effects depending on the target cell type and on the nature of the receptors expressed. IL-4 and IL-13 inhibit TNFα and IFNγ-mediated induction of antimicrobial peptides BD-2 and BD-3 by keratinocytes [25–27]. It has been suggested that the increased expression of IL-4 and IL-13 in atopic dermatitis skin may explain the susceptibility to bacterial and viral skin infections by reducing antimicrobial peptide expression [26, 27]. On the other hand, as TLR3 agonists , IL-4 or IL-13 synergizes with TNFα or IL-1β  to induce expression of thymic stromal lymphopoietin (TSLP) in keratinocytes and subsequently induces the maturation of CD11c+ dendritic cells involved in allergic inflammation. Besides regulation of their production, IL-4 and IL-13 activities can be modified by differential expression of their receptors. Indeed, IL-4 and IL-13 upregulate IL-13R2 expression in keratinocytes  whereas IFNγ or IL-13, but not IL-4, upregulates expression of IL-13R1 on keratinocytes in vitro .
IFNγ, mainly produced by the Th1 lymphocyte subset and by natural killer cells, is implicated in the regulation of different cellular processes such as antiviral responses, cell growth and differentiation, and immunoregulatory functions . IFNγ levels are increased in psoriatic skin, but not in acute AD lesions  reflecting an altered Th1/Th2 balance [33–35]. Interestingly, these IFN levels are reduced after antipsoriatic therapy [36, 37]. It is classically admitted that psoriasis is a Th1 disease. However, the coexistence of both IL-4-producing Th2 and IFN-γ-producing Th1 cells or Th1 dominance is observed in chronic AD lesion .
Several models strengthen the role of the members of the IL-1 family in inflammatory skin diseases. Transgenic mice constitutively expressing IL-1α in basal keratinocytes (under the control of the CK14 promoter) develop a spontaneous skin disease characterized by hair loss and inflammatory skin lesions displaying hyperkeratosis, acanthosis, parakeratosis, and a mononuclear cell infiltrate . In addition, IL-1RA deficient BALB/c mice develop arterial inflammation, arthritis, and a localized skin inflammation resembling human psoriasis . Recently, IL-36, α, β, and γ, 3 other members of the IL-1 family, have been found to be expressed in a psoriasis-like animal skin, as well as in the lesions of psoriatic patients .
TNFα is a multifunctional cytokine also produced by Th1 cells that mediates inflammation, immune response, and apoptosis. Biological activities are mediated by two distinct cell surface receptors: TNFR1 and TNFR2. Analysis of mice lacking either TNFR1 or TNFR2 has demonstrated that TNFR1 is critical for induction of skin inflammation by TNFα . Intradermal injection of TNFα leads to skin inflammation with elevated IL-6 production and induction of ICAM-1 expression by keratinocytes . TNFα is also involved in the production of antimicrobial peptide such as BD-2 and BD-3 [26, 42]. Extensive analysis of the transcriptional profile of keratinocytes treated with TNFα shows activation of innate and adaptive immune responses by inducing a large panel of chemokines that attracts neutrophils, macrophages, and T cells. The fundamental role played by TNFα in skin inflammation has been confirmed in mice lacking either IKK2 or IκBα, two molecules of the TNFα/NFκB signalling pathway. Finally, expression of TNFα and TNFR1 is increased in lesional psoriatic compared to normal skin . These in vitro and in vivo observations are confirmed by numerous clinical studies of successful anti-TNF therapy in psoriasis since the first report in 2000 [44–46]. In contrast, a pilot study on the effect of infliximab on 9 patients with moderate to severe AD was disappointed . Nevertheless, these cytokines do not seem to be involved in hyperplasia of epidermal keratinocytes observed in chronic skin lesions of AD and psoriasis. The hyperplasia and the alteration of barrier-related functions are linked to Th17 and Th22 cell subsets. Regarding the presence of Th17 lymphocytes in lesional psoriatic skin and the critical role of IL-17 in the pathogenesis of psoriasis [2, 8], presence and involvement of Th17 in AD has been evaluated. The percentage of Th17 cells was increased in peripheral blood of AD patients, in correlation with the severity of AD, and dermis IL-17-immunostaining was strongest in acute lesions when compared to the chronic one [48, 49]. Nograles et al. showed that the Th2 microenvironment has a suppressive effect on Th17 pathway by decreasing IL-17 receptor expression . Whatever the case, IL-17A expression in lesional AD is almost unchanged when compared to normal skin, whereas it is increased in psoriasis  (unpublished data). In addition, expression of IL-23p19, IL-12p40, but not IL-12p35, is increased in psoriatic skin [51–53] and IL-23p19 expression is associated with dendritic cells infiltrated in the skin .
As soon IL-22 has been described to target keratinocytes, it has been observed that the cytokine induces a “psoriasis-like” phenotype on RHE, that is, hyperplasia with a thickening of the spinous layer and a disappearance of the granular layer [6, 11], and that the expression and secretory patterns of IL-22-treated keratinocytes resembled most of the features of psoriatic lesions . Indeed, IL-22 is overexpressed in psoriatic lesions, whereas IL-22R1 and IL-10R2 are expressed at a similar level in psoriatic and healthy skin [37, 55]. T cells infiltrating psoriatic lesions are an important source of IL-22, higher than peripheral T cells from psoriatic patients or controls . Further analysis of T cell infiltrating psoriatic skin showed IL-17 and IL-22 coproduction by Th17 lymphocytes [56, 57]. Innate sources of IL-17 have been also considered in skin. Recently, a dermal RORγt γδ T cells producing IL-17 following exposure to IL-1β plus IL-23 have been described [58, 59]. In psoriatic patients, these γδ Tcells were greatly increased in affected skin and produced large amounts of IL-17 . Attention could also be paid to RORγt+, CCR6+, CD4neg, NK1.1neg iNKT cells  as a potential source of IL-17 in psoriasis, since these cells have been also described in skin and draining lymph nodes and respond to inflammatory signals in mice .
Beside the Th17 cells, a so-called Th22 subpopulation producing IL-22 but nor IL-17 or IFNγ has been recently described in circulation and in normal human dermis, expressing CCR6 and the skin-homing receptors CCR4 and CCR10 [62–64]. Th22 clones derived from psoriatic lesional skin have been further described, mostly in the epidermis compartment of the skin . Not specific of psoriasis, Th22 are also increased in lesional skin of atopic dermatitis and a correlation is observed between the number of Th22 cells and disease severity . In the chronic phase of AD, Th22 subset is induced by Langerhans cells  along with a Th1 cell response believed to be induced by IDCs. Otherwise, both in murine and human, it has been suspected that non-T cells in skin produce IL-22, such as dendritic cells, NK cells, macrophages, and so forth. [68, 69]. In our hands, IL-22 expression in lesional AD and psoriasis was enhanced when compared to normal skin  (submitted results). Interestingly, rare patients with simultaneous occurrence of psoriasis and AD have been recently described . Predominant antigen (Ag)-specific Th1 and Th17 lymphocytes infiltrates psoriatic lesions whereas Th2 infiltrates AD lesions, and IL-22 are detected in both lesions. Taken together, it appears that psoriasis is overall Th1 and Th17 mediated and AD is overall mediated by Th2 and Th22 cells, and that Th1 cells also contribute to the chronic phase of AD. In most cases, psoriasis and AD are mutually exclusive for individuals, but in rare cases can be concomitants, in which case Ag-specific T cells determine the specific pathogenesis .
4. Modelization of AD versus Psoriasis by Specific Sets of Cytokines
Taking into account (a) the preferential cytokine environment in AD vs psoriatic lesions and (b) those of these cytokines able to target keratinocytes , we design in vitro experiments aiming to culture NHEK and RHE with an IL-22, TNFα, IL-4, and IL-13 mix or an IL-22, TNFα, and IL-17 mix, with the objective to, respectively, mimic AD and psoriasis epidermis. In these models, we finally analyzed the effects of the change from IL-17 (Th17, psoriasis) to IL-4/IL-13 (Th2, AD), in the same cytokine background (TNFα and IL-22). We hypothesize that these cytokines are not only specific biomarkers of the diseases, but are responsible for the phenotypes and the molecular signatures of these diseases.
The overall response of human keratinocyte monolayers (NHEK) to defined cytokines aiming to mimic AD or psoriasis environment was analyzed using Affymetrix technology, covering almost all the human transcriptome (Figure 1). In the representative experiment reported, among more than 20 000 individual genes analyzed, about 750–800 genes were down- or upregulated by more than 2-fold by each treatment. These results are indicative; it should be noted that this model is a “picture” of the transcriptome of these keratinocytes under defined culture conditions, at a given time (24 h); many additional genes should have been detected under additional experimental conditions. Taking into account these remarks, we expected from these models a representative approach of a global physiopathological situation.
Interestingly, there is a large overlap of more than 60% between the two treatments. Among these modulated genes, are many epidermis differentiation-related genes, such as filaggrin (Figure 2), which is known to be strongly downregulated by IL-22 (present in the two cytokine mixtures), alone or in combinations . Most of the so-called epidermis differentiation markers are downregulated in NHEK by both AD and PSO NHEK treatments, as observed in pathologic skin. However, some of these markers such as genes from the small prolin rich proteins (SPRRs) family are differentially expressed in AD and PSO conditions. For example, SPRR2A is strongly overexpressed in the IL-17-containing mix (PSO) and downregulated in the IL-4/IL-13-containing mix (AD) (Figure 2).
In addition to differentiation markers, many other genes are commonly modulated by both cytokine mixes and TNFα by itself also strongly contribute to the “common” stimulation through. On the other hand, great differences between the two stimuli are observed for the antimicrobial peptides S100A7/psoriasin and BD-2; (Figure 2). S100A7 is dramatically overexpressed by the IL-17-containing mix (PSO) when compared to the weak enhancement induced by the IL-4/IL-13-containing mix (AD). According to its background expression in nonstimulated NHEK, BD-2 is dramatically enhanced in PSO model, and almost unmodified following AD stimulation. In contrast, the IL-13RA2 is overexpressed in the IL-4/IL-13-containing mix (AD) (Figure 2). These results parallel the in vivo situation, in which psoriatic skin is characterized by high expression levels of these antimicrobial peptides, functionally associated to a large degree of resistance to infections in psoriasis, and exacerbated infection sensitivity in AD skin .
A large number of chemokines are also produced, as indicated in Table 1. Whereas a chemokine such as CCL20 expression is more induced by the PSO mix as expected, CXCL10 is overexpressed only by the AD mix.
Since NHEK are undifferentiated primary keratinocytes cultured as monolayers, we further transpose the study to a more relevant epidermis model, reconstructed human epidermis (RHE), which closely resembles human skin epidermis (Figure 3). RHE are cornified 3D epithelia emerging from NHEK cultured at the air-medium interface . The treatment of RHE with DA mix for 2 days led to significant modification of the histology of the epidermis, with an apparent loss of cohesion of the tissue, and orthokeratosis. The replacement of the Th2 cytokines by IL-17 (PSO model) leads to more dramatic tissue damage with destabilization of the epidermis, small pycnotic cells associated to parakeratosis, and a tendency to the disappearance of the granular layer. These in vitro phenotypes, respectively, parallel histology of AD and psoriasis lesions . The decrease of expression of adhesion molecules could account for this phenotype. Clearly expressed in the epidermal layer of control RHE (Figure 3) as in normal skin, the treatment for only 48 h of these tissues by the two cytokine mixes resulted in a clear decrease in filaggrin accumulation. We previously showed that IL-22, present in the 2 mixes, decrease filaggrin expression . As this effect was stronger in the AD mix, we suggested that it is due to an additive effect of IL-4/IL-13.
Regarding the antimicrobial peptides, we observed a local expression of S100A7 in untreated RHE, a strong overexpression following AD treatment and, as expected, a dramatic expression with the PSO mix. The BD-2 protein is neither detected in “healthy” RHE nor in the AD model, but was strongly induced by the PSO mix. This huge enhancement results from the synergistic effect of IL-17 added to the IL-22/TNFα combination. We demonstrated that the immununohistological variations of filaggrin, S100A7, and BD-2 are confirmed at the mRNA level (Figure 4).
Taken together, these different patterns of in vitro expression were also found in chronic AD and psoriatic skin lesions. Interestingly, we demonstrate that by using exclusively keratinocytes and sets of cytokines, in the absence of other epidermis or dermis skin cells, resident or infiltrating immune cells, we can offer in vitro models that approach AD and psoriasis skin lesions. Obviously, this is downstream events that we model with keratinocytes and cytokines, depending of upstream mechanisms of recruitment and activation of other innate adaptive immune cells. Anyway, cytokine are sufficient to induce phenotypic features of chronic AD and psoriasis on keratinocytes. If still necessary, it highlights the fact that specific cytokines or combinations of cytokines are key targets for biotherapies. This approach is already largely developed for psoriasis treatments, not so far for AD treatments. Whatever, the effect of combination of cytokine are complex to analyze, and also depends on other parameters such as dose or kinetic of expression. Of course these simple keratinocyte models can be improved, but however, one lesson from these experiments is that on a common background, the presence of one additional cytokine can shift the tissue response toward a given phenotype, mutually exclusive such as in AD and psoriasis.
5. Concluding Remarks
In recent years, an increasing number of reports have led to define a panel of proinflammatory cytokines able to play a central role in the induction and maintenance of chronic skin inflammation. The presence of specific subsets of Th lymphocytes polarizes the response, and keratinocyte are downstream targets of these cytokines. This cytokine environment plays a central role on the skin morphology and innate immunity, giving to keratinocytes a status of bona fide actors of the innate immunity.
The specific cytokine environment in chronic AD includes an IL-22, TNFα, IL-4, and IL-13 specific combination. Altogether, these cytokines are able to mimic an “atopic dermatitis like” state on NHEK and RHE. IL-22 is especially involved in the development of the epidermal hyperplasia and hypogranulosis. TNFα is especially involved in the induction of the innate response, in synergy with IL-22, whereas IL-4 and IL-13 inhibits the BD-2 production. A large number of chemokines are also produced. Despite the production of neutrophil-attractive chemokines, the absence of neutrophils in AD lesions remains to be further studied. In psoriasis lesions, over expression of IL-17 is observed whereas IL-4 and IL-13 were not detected. Interestingly, the replacement of IL-4 and IL-13 by IL-17 from this cocktail is able to mimic in vitro a “psoriasis-like” status on keratinocytes.
These studies open the way to new therapeutic strategies focusing on more specific and downstream targets, that is, keratinocytes targeting cytokines rather than on a systemic inhibition of T lymphocytes overproducing cytokines. The specific blockade of new cytokines or their receptors is an alternative approach for the treatment of AD. In any case, cytokines are very potent factors, with the advantages (or the defects, depending on the case) to be pleiotropic and redundant, and to be involved in cascade. Such treatment could reduce the biological effects of an overproduced cytokine and/or disrupt the vicious circle involving it. Targeting more than one cytokine could be a valuable strategy to assure a complete and sustained clinical improvement.
Obviously, the in vitro models of inflammatory epidermis induced by a specific set of cytokines could be useful tools to screen new drugs.
- P. Di Meglio, K. Gayathri Perera, and F. O. Nestle, “The multitasking organ: recent insights into skin immune function,” Immunity, vol. 35, no. 6, pp. 857–869, 2011.
- E. Guttman-Yassky, K. E. Nograles, and J. G. Krueger, “Contrasting pathogenesis of atopic dermatitis and psoriasis-part II: immune cell subsets and therapeutic concepts,” Journal of Allergy and Clinical Immunology, vol. 127, no. 6, pp. 1420–1432, 2011.
- E. Guttman-Yassky, K. E. Nograles, and J. G. Krueger, “Contrasting pathogenesis of atopic dermatitis and psoriasis-part I: clinical and pathologic concepts,” Journal of Allergy and Clinical Immunology, vol. 127, no. 5, pp. 1110–1118, 2011.
- C. N. A. Palmer, A. D. Irvine, A. Terron-Kwiatkowski et al., “Common loss-of-function variants of the epidermal barrier protein filaggrin are a major predisposing factor for atopic dermatitis,” Nature Genetics, vol. 38, no. 4, pp. 441–446, 2006.
- J. Lecron, F. Morel, and K. Boniface, “Keratinocytes as targets for cytokines in skin inflammation,” in Recent Advances in Skin Immunology, S. Saeland, Ed., p. 46, Kerala, Kerala, India, 2008.
- K. Boniface, F. X. Bernard, M. Garcia, A. L. Gurney, J. C. Lecron, and F. Morel, “IL-22 inhibits epidermal differentiation and induces proinflammatory gene expression and migration of human keratinocytes,” Journal of Immunology, vol. 174, no. 6, pp. 3695–3702, 2005.
- K. Boniface, C. Diveu, F. Morel et al., “Oncostatin M secreted by skin infiltrating T lymphocytes is a potent keratinocyte activator involved in skin inflammation,” Journal of Immunology, vol. 178, no. 7, pp. 4615–4622, 2007.
- N. J. Wilson, K. Boniface, J. R. Chan et al., “Development, cytokine profile and function of human interleukin 17-producing helper T cells,” Nature Immunology, vol. 8, no. 9, pp. 950–957, 2007.
- R. Caruso, E. Botti, M. Sarra et al., “Involvement of interleukin-21 in the epidermal hyperplasia of psoriasis,” Nature Medicine, vol. 15, no. 9, pp. 1013–1015, 2009.
- M. Wittmann, R. Purwar, C. Hartmann, R. Gutzmer, and T. Werfel, “Human keratinocytes respond to interleukin-18: implication for the course of chronic inflammatory skin diseases,” Journal of Investigative Dermatology, vol. 124, no. 6, pp. 1225–1233, 2005.
- K. Boniface, J. C. Lecron, F. X. Bernard et al., “Keratinocytes as targets for interleukin-10-related cytokines: a putative role in the pathogenesis of psoriasis,” European Cytokine Network, vol. 16, no. 4, pp. 309–319, 2005.
- S. M. Sa, P. A. Valdez, J. Wu 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,” Journal of Immunology, vol. 178, no. 11, pp. 2229–2240, 2007.
- R. Caruso, E. Botti, M. Sarra et al., “Involvement of interleukin-21 in the epidermal hyperplasia of psoriasis,” Nature Medicine, vol. 15, no. 9, pp. 1013–1015, 2009.
- K. Guilloteau, I. Paris, N. Pedretti et al., “Skin inflammation induced by the synergistic action of IL-17A, IL-22, oncostatin M, IL-1α, and TNF-α recapitulates some features of psoriasis,” Journal of Immunology, vol. 184, no. 9, pp. 5263–5270, 2010.
- Y. Carrier, H. L. Ma, H. E. Ramon et al., “Inter-regulation of Th17 cytokines and the IL-36 cytokines in vitro and in vivo: implications in psoriasis pathogenesis,” Journal of Investigative Dermatology, vol. 131, pp. 2428–2437, 2011.
- M. C. Lebre, A. M. G. Van Der Aar, L. Van Baarsen et al., “Human keratinocytes express functional toll-like receptor 3, 4, 5, and 9,” Journal of Investigative Dermatology, vol. 127, no. 2, pp. 331–341, 2007.
- L. S. Miller, “Toll-like receptors in skin,” Advances in Dermatology C, vol. 24, pp. 71–87, 2008.
- H. Kinoshita, T. Takai, T. Anh Le et al., “Cytokine milieu modulates release of thymic stromal lymphopoietin from human keratinocytes stimulated with double-stranded RNA,” Journal of Allergy and Clinical Immunology, vol. 123, no. 1, pp. 179–186, 2009.
- K. E. Nograles, L. C. Zaba, E. Guttman-Yassky et al., “Th17 cytokines interleukin (IL)-17 and IL-22 modulate distinct inflammatory and keratinocyte-response pathways,” British Journal of Dermatology, vol. 159, no. 5, pp. 1092–1102, 2008.
- M. B. M. Teunissen, C. W. Koomen, R. De Waal Malefyt, E. A. Wierenga, and J. D. Bos, “Interleukin-17 and interferon-γ synergize in the enhancement of proinflammatory cytokine production by human keratinocytes,” Journal of Investigative Dermatology, vol. 111, no. 4, pp. 645–649, 1998.
- C. Albanesi, A. Cavani, and G. Girolomoni, “IL-17 is produced by nickel-specific T lymphocytes and regulates ICAM-1 expression and chemokine production in human keratinocytes: synergistic or antagonist effects with IFN-γ and TNF-α,” Journal of Immunology, vol. 162, no. 1, pp. 494–502, 1999.
- S. C. Liang, X. Y. Tan, D. P. Luxenberg et al., “Interleukin (IL)-22 and IL-17 are coexpressed by Th17 cells and cooperatively enhance expression of antimicrobial peptides,” Journal of Experimental Medicine, vol. 203, no. 10, pp. 2271–2279, 2006.
- S. Ghannam, C. Dejou, N. Pedretti et al., “CCL20 and β-defensin-2 induce arrest of human Th17 cells on inflamed endothelium in vitro under flow conditions,” Journal of Immunology, vol. 186, no. 3, pp. 1411–1420, 2011.
- R. Purwar, T. Werfel, and M. Wittmann, “IL-13-stimulated human keratinocytes preferentially attract CD4+ CCR4+ T cells: possible role in atopic dermatitis,” Journal of Investigative Dermatology, vol. 126, no. 5, pp. 1043–1051, 2006.
- C. Albanesi, H. R. Fairchild, S. Madonna et al., “IL-4 and IL-13 negatively regulate TNF-β- and IFN-γ-induced β-defensin expression through STAT-6, suppressor of cytokine signaling (SOCS)-1, and SOCS-3,” Journal of Immunology, vol. 179, no. 2, pp. 984–992, 2007.
- I. Nomura, E. Goleva, M. D. Howell et al., “Cytokine milieu of atopic dermatitis, as compared to psoriasis, skin prevents induction of innate immune response genes,” Journal of Immunology, vol. 171, no. 6, pp. 3262–3269, 2003.
- P. Y. Ong, T. Ohtake, C. Brandt et al., “Endogenous antimicrobial peptides and skin infections in atopic dermatitis,” New England Journal of Medicine, vol. 347, no. 15, pp. 1151–1160, 2002.
- S. I. Bogiatzi, I. Fernandez, J. C. Bichet et al., “Cutting edge: proinflammatory and Th2 cytokines synergize to induce thymic stromal lymphopoietin production by human skin keratinocytes,” Journal of Immunology, vol. 178, no. 6, pp. 3373–3377, 2007.
- M. David, D. Ford, J. Bertoglio, A. L. Maizel, and J. Pierre, “Induction of the IL-13 receptor α2-chain by IL-4 and IL-13 in human keratinocytes: involvement of STAT6, ERK and p38 MAPK pathways,” Oncogene, vol. 20, no. 46, pp. 6660–6668, 2001.
- J. Wongpiyabovorn, H. Suto, H. Ushio et al., “Up-regulation of interleukin-13 receptor α1 on human keratinocytes in the skin of psoriasis and atopic dermatitis,” Journal of Dermatological Science, vol. 33, no. 1, pp. 31–40, 2003.
- M. A. Farrar and R. D. Schreiber, “The molecular cell biology of interferon-γ and its receptor,” Annual Review of Immunology, vol. 11, pp. 571–611, 1993.
- I. Nomura, B. Gao, M. Boguniewicz, M. A. Darst, J. B. Travers, and D. Y. M. Leung, “Distinct patterns of gene expression in the skin lesions of atopic dermatitis and psoriasis: a gene microarray analysis,” Journal of Allergy and Clinical Immunology, vol. 112, no. 6, pp. 1195–1202, 2003.
- K. Uyemura, M. Yamamura, D. F. Fivenson, R. L. Modlin, and B. J. Nickoloff, “The cytokine network in lesional and lesion-free psoriatic skin is characterized by a T-helper Type 1 cell-mediated response,” Journal of Investigative Dermatology, vol. 101, no. 5, pp. 701–705, 1993.
- A. Szegedi, M. Aleksza, A. Gonda et al., “Elevated rate of Thelper1 (TH1) lymphocytes and serum IFN-γ levels in psoriatic patients,” Immunology Letters, vol. 86, no. 3, pp. 277–280, 2003.
- L. M. Austin, M. Ozawa, T. Kikuchi, I. B. Walters, and J. G. Krueger, “The majority of epidermal T cells in psoriasis vulgaris lesions can produce type 1 cytokines, interferon-γ, interleukin-2, and tumor necrosis factor-α, defining TC1 (cytotoxic T lymphocyte) and TH1 effector populations: a type 1 differentiation bias is also measured in circulating blood T cells in psoriatic patients,” Journal of Investigative Dermatology, vol. 113, no. 5, pp. 752–759, 1999.
- W. L. Trepicchio, M. Ozawa, I. B. Walters et al., “Interleukin-11 therapy selectively downregulates type I cytokine proinflammatory pathways in psoriasis lesions,” Journal of Clinical Investigation, vol. 104, no. 11, pp. 1527–1537, 1999.
- K. Wolk, E. Witte, E. Wallace et al., “IL-22 regulates the expression of genes responsible for antimicrobial defense, cellular differentiation, and mobility in keratinocytes: a potential role in psoriasis,” European Journal of Immunology, vol. 36, no. 5, pp. 1309–1323, 2006.
- M. Grewe, C. A. F. M. Bruijnzeel-Koomen, E. Schöpf et al., “A role for Th1 and Th2 cells in the immunopathogenesis of atopic dermatitis,” Immunology Today, vol. 19, no. 8, pp. 359–361, 1998.
- R. W. Groves, H. Mizutani, J. D. Kieffer, and T. S. Kupper, “Inflammatory skin disease in transgenic mice that express high levels of interleukin 1α in basal epidermis,” Proceedings of the National Academy of Sciences of the United States of America, vol. 92, no. 25, pp. 11874–11878, 1995.
- J. Shepherd, M. C. Little, and M. J. H. Nicklin, “Psoriasis-like cutaneous inflammation in mice lacking interleukin-1 receptor antagonist,” Journal of Investigative Dermatology, vol. 122, no. 3, pp. 665–669, 2004.
- S. Kondo and D. N. Sauder, “Tumor necrosis factor (TNF) receptor type 1 (p55) is a main mediator for TNF-α-induced skin inflammation,” European Journal of Immunology, vol. 27, no. 7, pp. 1713–1718, 1997.
- J. Harder, J. Bartels, E. Christophers, and J. M. Schröder, “Isolation and characterization of human β-defensin-3, a novel human inducible peptide antibiotic,” Journal of Biological Chemistry, vol. 276, no. 8, pp. 5707–5713, 2001.
- P. Ettehadi, M. W. Greaves, D. Wallach, D. Aderka, and R. D. R. Camp, “Elevated tumour necrosis factor-alpha (TNF-α) biological activity in psoriatic skin lesions,” Clinical and Experimental Immunology, vol. 96, no. 1, pp. 146–151, 1994.
- C. J. Oh, K. M. Das, and A. B. Gottlieb, “Treatment with anti-tumor necrosis factor α (TNF-α) monoclonal antibody dramatically decreases the clinical activity of psoriasis lesions,” Journal of the American Academy of Dermatology, vol. 42, no. 5, pp. 829–830, 2000.
- U. Chaudhari, P. Romano, L. D. Mulcahy, L. T. Dooley, D. G. Baker, and A. B. Gottlieb, “Efficacy and safety of infliximab monotherapy for plaque-type psoriasis: a randomised trial,” The Lancet, vol. 357, no. 9271, pp. 1842–1847, 2001.
- C. L. Leonardi, J. L. Powers, R. T. Matheson et al., “Etanercept as monotherapy in patients with psoriasis,” New England Journal of Medicine, vol. 349, no. 21, pp. 2014–2022, 2003.
- A. Jacobi, C. Antoni, B. Manger, G. Schuler, and M. Hertl, “Infliximab in the treatment of moderate to severe atopic dermatitis.,” Journal of the American Academy of Dermatology, vol. 52, no. 3, pp. 522–526, 2005.
- C. Koga, K. Kabashima, N. Shiraishi, M. Kobayashi, and Y. Tokura, “Possible pathogenic role of Th17 cells for atopic dermatitis,” Journal of Investigative Dermatology, vol. 128, no. 11, pp. 2625–2630, 2008.
- M. Toda, D. Y. M. Leung, S. Molet et al., “Polarized in vivo expression of IL-11 and IL-17 between acute and chronic skin lesions,” Journal of Allergy and Clinical Immunology, vol. 111, no. 4, pp. 875–881, 2003.
- K. E. Nograles, M. Suárez-Fariñas, A. Shemer et al., “Atopic dermatitis keratinocytes exhibit normal TH17 cytokine responses,” Journal of Allergy and Clinical Immunology, vol. 125, no. 3, pp. 744–e2, 2010.
- J. R. Chan, W. Blumenschein, E. Murphy et al., “IL-23 stimulates epidermal hyperplasia via TNF and IL-20R2-dependent mechanisms with implications for psoriasis pathogenesis,” Journal of Experimental Medicine, vol. 203, no. 12, pp. 2577–2587, 2006.
- G. Piskin, R. M. R. Sylva-Steenland, J. D. Bos, and M. B. M. Teunissen, “In vitro and in situ expression of IL-23 by keratinocytes in healthy skin and psoriasis lesions: enhanced expression in psoriatic skin,” Journal of Immunology, vol. 176, no. 3, pp. 1908–1915, 2006.
- E. Lee, W. L. Trepicchio, J. L. Oestreicher et al., “Increased expression of interleukin 23 p19 and p40 in lesional skin of patients with psoriasis vulgaris,” Journal of Experimental Medicine, vol. 199, no. 1, pp. 125–130, 2004.
- A. M. Bowcock and J. G. Krueger, “Getting under the skin: the immunogenetics of psoriasis,” Nature Reviews Immunology, vol. 5, no. 9, pp. 699–711, 2005.
- K. Boniface, E. Guignouard, N. Pedretti et al., “A role for T cell-derived interleukin 22 in psoriatic skin inflammation,” Clinical and Experimental Immunology, vol. 150, no. 3, pp. 407–415, 2007.
- M. A. Lowes, T. Kikuchi, J. Fuentes-Duculan et al., “Psoriasis vulgaris lesions contain discrete populations of Th1 and Th17 T cells,” Journal of Investigative Dermatology, vol. 128, no. 5, pp. 1207–1211, 2008.
- J. Pène, S. Chevalier, L. Preisser et al., “Chronically inflamed human tissues are infiltrated by highly differentiated Th17 lymphocytes,” Journal of Immunology, vol. 180, no. 11, pp. 7423–7430, 2008.
- E. E. Gray, K. Suzuki, and J. G. Cyster, “Cutting edge: identification of a motile IL-17-producing γδ T cell population in the dermis,” Journal of Immunology, vol. 186, no. 11, pp. 6091–6095, 2011.
- Y. Cai, X. Shen, C. Ding et al., “Pivotal role of dermal IL-17-producing gammadelta T cells in skin inflammation,” Immunity, vol. 35, no. 4, pp. 596–610, 2011.
- M. L. Michel, A. C. Keller, C. Paget et al., “Identification of an IL-17-producing NK1.1neg iNKT cell population involved in airway neutrophilia,” Journal of Experimental Medicine, vol. 204, no. 5, pp. 995–1001, 2007.
- J. M. Doisne, C. Becourt, L. Amniai et al., “Skin and peripheral lymph node invariant NKT cells are mainly retinoic acid receptor-related orphan receptor γt+and respond preferentially under inflammatory conditions,” Journal of Immunology, vol. 183, no. 3, pp. 2142–2149, 2009.
- K. E. Nograles, L. C. Zaba, E. Guttman-Yassky et al., “Th17 cytokines interleukin (IL)-17 and IL-22 modulate distinct inflammatory and keratinocyte-response pathways,” British Journal of Dermatology, vol. 159, no. 5, pp. 1092–1102, 2008.
- S. Trifari, C. D. Kaplan, E. H. Tran, N. K. Crellin, and H. Spits, “Identification of a human helper T cell population that has abundant production of interleukin 22 and is distinct from TH-17, TH1 and TH2 cells,” Nature Immunology, vol. 10, no. 8, pp. 864–871, 2009.
- T. Duhen, R. Geiger, D. Jarrossay, A. Lanzavecchia, and F. Sallusto, “Production of interleukin 22 but not interleukin 17 by a subset of human skin-homing memory T cells,” Nature Immunology, vol. 10, no. 8, pp. 857–863, 2009.
- S. Eyerich, K. Eyerich, D. Pennino et al., “Th22 cells represent a distinct human T cell subset involved in epidermal immunity and remodeling,” Journal of Clinical Investigation, vol. 119, no. 12, pp. 3573–3585, 2009.
- K. E. Nograles, L. C. Zaba, A. Shemer et al., “IL-22-producing "T22" T cells account for upregulated IL-22 in atopic dermatitis despite reduced IL-17-producing TH17 T cells,” Journal of Allergy and Clinical Immunology, vol. 123, no. 6, pp. 1244–e2, 2009.
- H. Fujita, K. E. Nograles, T. Kikuchi, J. Gonzalez, J. A. Carucci, and J. G. Krueger, “Human Langerhans cells induce distinct IL-22-producing CD4+ T cells lacking IL-17 production,” Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 51, pp. 21795–21800, 2009.
- P. C. M. Res, G. Piskin, O. J. de Boer et al., “Overrepresentation of IL-17A and IL-22 Producing CD8 T Cells in Lesional Skin Suggests Their Involvement in the pathogenesis of psoriasis,” PLoS ONE, vol. 5, no. 11, Article ID e14108, 2010.
- A. B. Van Belle, M. de Heusch, M. M. Lemaire et al., “IL-22 is required for imiquimod-induced psoriasiform skin inflammation in mice,” Journal of Immunology, vol. 188, no. 1, pp. 462–469, 2012.
- S. Eyerich, A. T. Onken, S. Weidinger et al., “Mutual antagonism of T cells causing psoriasis and atopic eczema,” New England Journal of Medicine, vol. 365, no. 3, pp. 231–238, 2011.
- M. Rosdy, B. Bertino, V. Butet, S. Gibbs, M. Ponec, and M. Darmon, “Retinoic Acid inhibits epidermal differentiation when applied topically on the stratum corneum of epidermis formed in vitro by human keratinocytes grown on defined medium,” In Vitro and Molecular Toxicology, vol. 10, no. 1, pp. 39–47, 1997.