interleukin-8 and Psoriasis

The excellent clinical efficacy of anti-interleukin 17A (IL-17A) biologics on psoriasis indicates a crucial pathogenic role of IL-17A in this autoinflammatory skin disease. IL-17A accelerates the proliferation of epidermal keratinocytes. Keratinocytes produce a myriad of antimicrobial peptides and chemokines, such as CXCL1, CXCL2, CXCL8, and CCL20. Antimicrobial peptides enhance skin inflammation. IL-17A is capable of upregulating the production of these chemokines and antimicrobial peptides in keratinocytes. CXCL1, CXCL2, and CXCL8 recruit neutrophils and CCL20 chemoattracts IL-17A-producing CCR6

Topics: Cell Proliferation; Chemokine CCL20; Chemokine CXCL1; Chemokine CXCL2; Epidermis; Humans; Interleukin-17; Interleukin-8; Keratinocytes; Neutrophils; Psoriasis

T helper (Th) 17 cells have crucial functions in host defense, and dysregulated Th17 responses mediate a variety of autoimmune and inflammatory conditions. Th17 cells coexpress interleukin (IL)-22, and its receptor is expressed on epidermal keratinocytes. IL-17 and IL-22 cooperatively enhance some immunological responses. A close relationship between IL-17 and the cutaneous milieu has been suggested by a number of observations. IL-17 induces the production of certain cytokines, chemokines and antimicrobial peptides by keratinocytes, and its cooperation with IL-22 has been documented. Recent findings have suggested that Th17 cells profoundly participate in the pathogenesis of certain skin disorders, in particular, psoriasis. The concept of the subsets of T cells responsible for psoriasis has been modified in the order of Th1, T cytotoxic 1, and again Thl, and Thl7 cells. IL-22 is the strongest cytokine in the keratinocyte-proliferative ability. Since IL-22 is produced by Th17 cells, they are crucial for the proliferation of keratinocytes. Furthermore, IL-22 with the help of IL-17 can induce the critical events of psoriasis, including signal transducer and activator of transcription 3 (STAT3) activation, cytokine/chemokine (IL-8 etc.) production, and antimicrobial peptide elaboration. For maintaining Th17 cells, IL-23 is required and is released from tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthetase (iNOS)-producing dendritic cells (TIP-DCs). TIP-DCs are activated via an autocrine mechanism by virtue of TNF-alpha. The above cytokine network in the pathogenesis of psoriasis has been proven by the therapeutic effectiveness of cytokine-blocking biologics. Antibodies against TNF-alpha or its soluble receptor have already been widely used in the treatment of psoriasis. The involvement of Th17 cells has also been shown in allergen-specific immune responses. The percentage of Th17 cells is increased in the peripheral blood of patients with atopic dermatitis (AD) and associated with the severity of AD. Drug eruption is another disease where Th17 cells are involved in the pathogenesis. The percentage of circulating Th17 cells are increased in drug-induced hypersensitivity syndrome, etc. Th17 cells and IL-22 are increased in patients with acute generalized exanthematous pustulosis. Since IL-17 and IL-22 cooperatively stimulate keratinocytes to produce IL-8, keratinocyte-derived IL-8 contributes to the accumulation ofneutrophils in the lesi

Topics: Antimicrobial Cationic Peptides; Autocrine Communication; Autoimmune Diseases; Dendritic Cells; Dermatitis, Atopic; Drug Eruptions; Humans; Interleukin-17; Interleukin-22; Interleukin-23; Interleukin-8; Interleukins; Keratinocytes; Nitric Oxide Synthase Type II; Psoriasis; Receptors, Interleukin; STAT3 Transcription Factor; Th17 Cells; Tumor Necrosis Factor-alpha

The purpose of the present review is to update knowledge on acute generalized exanthematous pustulosis (AGEP) in terms of epidemiology, pathogenesis, cause, clinical features, diagnosis, and treatment.. AGEP is a rare reaction pattern attributed mainly to drugs. Drug-specific T cells (CD4+ and CD8+) and the production of interleukin-8/CXCL8 play an important role in its pathogenesis. A large-scale case-control study (EuroSCAR study) revealed a broad spectrum of drugs strongly associated with AGEP characterized by different time patterns (latent periods). Recent publications have supported the recognized role of individual drugs in the induction of AGEP and some have reported newly incriminated drugs. Many recent publications on AGEP have used the AGEP validation score (EuroSCAR group criteria) to establish the diagnosis. The value of in-vivo tests (mainly patch tests), in-vitro tests (the lymphocyte transformation test and cytokine release tests), or both for the identification of causative drugs has been demonstrated. Infections do not play a prominent role in the development of AGEP. There is no evidence for the assumption that AGEP is a variant of pustular psoriasis. Unique observations related to AGEP include a marked female predominance, a possible role for seasonality and a causal role for spider bites.. A broad spectrum of drugs is associated with AGEP, a T cell-mediated reaction. Genetic susceptibility and the possible role of other risk factors in AGEP should be further evaluated in larger studies of AGEP patients with a validated diagnosis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Drug Hypersensitivity; Exanthema; Female; Humans; Interleukin-8; Lymphocyte Activation; Patch Tests; Psoriasis; Sex Factors; Spider Bites; Spiders

Topics: Arthritis, Rheumatoid; Behcet Syndrome; Biomarkers; Bronchitis; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-10; Interleukin-7; Interleukin-8; Interleukin-9; Lupus Erythematosus, Systemic; Psoriasis; Reagent Kits, Diagnostic; Sjogren's Syndrome

Topics: Autoimmune Diseases; Drugs, Investigational; Humans; Interleukin-8; Psoriasis; T-Lymphocytes

Interleukin-8 (IL-8) is a potent chemotactic and proinflammatory cytokine, produced in the skin by a variety of cells in response to inflammatory stimuli. Recent studies suggest that in addition to its potent actions on leukocytes, IL-8 exerts a direct influence on several functions of human epidermal cells such as chemotaxis, Candida albicans killing activity or proliferation. The effects of IL-8 are mediated by binding to two types of specific high-affinity receptors which contain seven transmembrane domains typical of guanine nucleotide binding protein (G protein)-coupled receptors. In the skin, a broad spectrum of cells such as neutrophils, T lymphocytes, mast cells, dermal macrophages, endothelial cells and keratinocytes possess binding sites for IL-8. Recently, increased expression of epidermal IL-8 receptors has been observed in psoriasis an inflammatory and hyperproliferative skin disease. Since the effects of IL-8 may be modulated at the receptor level, the pharmacological manipulation of the IL-8 receptor may prove an important target for the therapy of skin diseases with increased IL-8 levels.

Topics: Binding Sites; Humans; Interleukin-8; Psoriasis; Receptors, Interleukin; Receptors, Interleukin-8A; Skin; Skin Physiological Phenomena

Psoriasis represents an inflammatory skin disorder which is characterized by a marked hyperproliferation of keratinocytes in association with vascular expansion, fibroblast activation, leukocyte infiltration, alterations of eicosanoid metabolism and of cytokine production. However, it is unclear at present whether these changes may be a cause or a result of the significantly increased keratinocyte turnover. More than one mechanism is involved in triggering active psoriasis, particularly a genetic predisposition and environmental factors affecting the immune system. Most of the therapeutic regimes used for the treatment of psoriasis are immunosuppressive. Therefore, it is tempting to speculate that a specific defect of the immune system represents an important pathogenic principle in psoriasis. There are several lines of evidence that changes in cytokine production by keratinocytes and immunocompetent cells in the skin of the patients (particularly of interleukin-6 and TGF-alpha) may play an important role in the propagation of the inflammatory response in psoriasis. Further studies are required to reveal the role of a local T-cell activation as a basic mechanism for initiation and maintenance of the psoriatic inflammatory response. Accordingly, parameters, such as the evaluation of cytokine production in vitro and in vivo, as well as the measurement of cellular activation products, may be useful tools for diagnosis and monitoring of psoriasis.

Topics: Autoimmune Diseases; Cytokines; Humans; Interferons; Interleukin-1; Interleukin-6; Interleukin-8; Lymphocyte Activation; Psoriasis; T-Lymphocytes; Tumor Necrosis Factor-alpha

Topics: Amino Acid Sequence; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Cytokines; Growth Substances; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Molecular Sequence Data; Neutrophils; Psoriasis

The constitutive production of interleukin-1 alpha-like material in normal human epidermis, its inflammatory properties, and the mechanism of its inflammatory action are briefly reviewed. The isolation of interleukin-8 from psoriatic lesions, its in vitro production, and leukocyte chemoattractant properties are also described. Available evidence suggests that interleukins-1 and -8 are inflammatory cytokines of major potential importance in the induction of leukocyte infiltrates in human skin.

Topics: Chemotactic Factors; Chemotaxis, Leukocyte; Epidermis; Humans; Interleukin-1; Interleukin-8; Psoriasis; Skin

Topics: Biological Factors; Chemotactic Factors; Cytokines; Humans; Interleukin-8; Neutrophils; Peptides; Psoriasis

Extracts of stratum corneum samples from psoriatic skin lesions have been shown to contain a group of neutrophil chemoattractant polypeptides, a major portion of which is distinct from C5a (des arg). One of the components may be identical to the novel monocyte-derived neutrophil chemotactic peptide which has recently been purified, sequenced and cloned. Synthesis of this agent may be induced by interleukin 1, this process possibly explaining part of the pro-inflammatory activity of interleukin 1 in the skin. These compounds may be important in the pathogenesis of inflammatory skin disease.

Topics: Chemotactic Factors; Chemotaxis, Leukocyte; Humans; Inflammation; Interleukin-1; Interleukin-8; Monocytes; Neutrophils; Psoriasis; Skin

To study the effect of Yinxieling decoction on PASI, TNF-α and IL-8 in patients with psoriasis vulgaris.. A total of 120 cases of psoriasis vulgaris were divided into 4 groups according to syndrome differentiation of TCM and randomized controlled method: wind heat syndrome group (group A), blood stasis syndrome group (group B), blood dryness syndrome group (group C) and control group (group D) (n=30 per group). Patients in observation groups were treated with Yinxieling decoction, while patients in control group were treated by placebo for 8 weeks. Levels of TNF-α and IL-8 were determined before treatment, 4 and 8 weeks after treatment. psoriasis area and severity index score was also performed before and after treatment.. psoriasis area and severity index score and serum level of TNF-α, IL-8 were significantly decreased in all groups. The decrease in three observation groups was more significant (P<0.05 or P<0.01), and the decrease in wind heat syndrome group was the most significant (P<0.01). psoriasis area and severity index was positively correlated with TNF-α and IL-8, respectively (P<0.05).. Yinxieling decoction has therapeutical effect on psoriasis vulgaris via regulating TNF-α and IL-8.

Topics: Adolescent; Adult; Aged; Drugs, Chinese Herbal; Female; Humans; Interleukin-8; Male; Middle Aged; Psoriasis; Severity of Illness Index; Treatment Outcome; Tumor Necrosis Factor-alpha; Young Adult

Tetradecylthioacetic acid (TTA) is a bioactive 3-thia fatty acid, giving hypolipidemic response, inhibiting the proliferation and increasing the differentiation of normal adult epidermal keratinocytes and showing anti-oxidant and anti-inflammatory effects. Psoriasis is an inflammatory disease associated with abnormalities in lipid profile, lipid peroxidation, antioxidant capacity, eicosanoid metabolism and increased frequency of cardiovascular events. On this background we have conducted a pilot study to explore the hypothesis that this modified fatty acid could improve dyslipidemia and reduce inflammation in psoriatic patients. In this double-blinded, placebo-controlled study, we assessed the metabolic effects of systemic TTA in a limited number of patients with mild to moderate psoriasis, 1000 mg TTA daily for 28 days. The most important findings were: (i) TTA reduced plasma total cholesterol, non HDL-cholesterol, LDL/HDL cholesterol ratio, triglycerides and total fatty acids; (ii) TTA decreased plasma TNF-α, IL-8 and VCAM-1; and (iii) plasma fatty acid composition changed with an increased level of monounsaturated fatty acids and decreased n-3 polyunsaturated fatty acids. In conclusion TTA exerts both hypolipidemic and anti-inflammatory effects in psoriasis patients. The results further indicate that TTA can be of therapeutic benefit for a subgroup of psoriatic patients.

Topics: Adult; Aged; Anti-Inflammatory Agents, Non-Steroidal; Double-Blind Method; Female; Humans; Hypolipidemic Agents; Interleukin-8; Lipids; Male; Middle Aged; Pilot Projects; Psoriasis; Sulfides; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1; Young Adult

TNF inhibitors have revolutionized the treatment of psoriasis vulgaris as well as psoriatic and rheumatoid arthritis and Crohn disease. Despite our understanding that these agents block TNF, their complex mechanism of action in disease resolution is still unclear.. To analyze globally the genomic effects of TNF inhibition in patients with psoriasis, and to compare genomic profiles of patients who responded or did not respond to treatment.. In a clinical trial using etanercept TNF inhibitor to treat psoriasis vulgaris (n = 15), Affymetrix gene arrays were used to analyze gene profiles in lesional skin at multiple time points during drug treatment (baseline and weeks 1, 2, 4, and 12) compared with nonlesional skin. Patients were stratified as responders (n = 11) or nonresponders (n = 4) on the basis of histologic disease resolution. Cluster analysis was used to define gene sets that were modulated with similar magnitude and velocity over time.. In responders, 4 clusters of downregulated genes and 3 clusters of upregulated genes were identified. Genes downmodulated most rapidly reflected direct inhibition of myeloid lineage immune genes. Upregulated genes included the stable dendritic cell population genes CD1c and CD207 (langerin). Comparison of responders and nonresponders revealed rapid downmodulation of innate IL-1beta and IL-8 sepsis cascade cytokines in both groups, but only responders downregulated IL-17 pathway genes to baseline levels.. Although both responders and nonresponders to etanercept inactivated sepsis cascade cytokines, response to etanercept is dependent on inactivation of myeloid dendritic cell genes and inactivation of the T(H)17 immune response.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Dendritic Cells; Down-Regulation; Etanercept; Gene Expression; Gene Expression Profiling; Humans; Immunoglobulin G; Interferon-gamma; Interleukin-17; Interleukin-1beta; Interleukin-8; Myeloid Cells; Oligonucleotide Array Sequence Analysis; Psoriasis; Receptors, Tumor Necrosis Factor; Signal Transduction; Skin; T-Lymphocytes, Helper-Inducer; Tumor Necrosis Factor-alpha; Up-Regulation

Psoriasis is one of the most frequent inflammatory skin diseases in which abnormal individual immune reactivity plays an important role. The aim of the present study was to describe selected immunological changes, concerning pro-inflammatory cytokines (TNF-alpha, IL-8) and adhesion molecules (sE-selectin, sP-selectin, sICAM-1), in 56 patients cured by Goeckerman's therapy (GT). GT includes dermal application of crude coal tar (containing polycyclic aromatic hydrocarbons) and exposure to UV radiation. When compared with the control group (healthy blood donors), the patients before GT had significantly increased serum levels of sE-selectin (p<0.001), sP-selectin (p<0.001), sICAM-1 (p<0.001) and IL-8 (p<0.001). Significantly decreased serum levels of sE-selectin (p<0.05) and significantly increased serum levels of IL-8 (p<0.05) were found after GT therapy. Serum levels of sICAM significantly correlated with the disease activity and with serum levels of sE-selectin. The level of PASI score (Psoriasis Area and Severity Index) significantly decreased after GT (p<0.001) and confirms the high efficiency GT. These findings confirmed that pro-inflammatory chemokine (IL-8) and adhesion molecules (sE-selectin, sP-selectin, sICAM-1) play an important role in the development and regulation of inflammation in psoriasis. Determination of sE-selectin and sICAM seems to be a promising marker of psoriasis's activity. Chemokine pathway (IL-8) and TNF-alpha activity seem to be modulated by Goeckerman's therapy (polycyclic aromatic hydrocarbons).

Topics: Adolescent; Adult; Aged; Biomarkers; Cell Adhesion Molecules; Coal Tar; Cytokines; E-Selectin; Female; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Keratolytic Agents; Male; Middle Aged; P-Selectin; Psoriasis; Severity of Illness Index; Time Factors; Treatment Outcome; Tumor Necrosis Factor-alpha; Ultraviolet Therapy

To seek for a better therapy for chronic plaque type psoriasis.. The effect of 35 patients in the tested group treated with combination therapy of composite Shendi Decoction (CSDD) and Diyin Tablet (DYT) was observed and compared with the effect of those treated by CSDD alone (25 cases) or by DYT alone (18 cases) as the control groups.. The total effective rate in the treated group was 94.29%, and that in the two control groups was 88.00% and 83.33% respectively. The markedly effective rate in the treated group was 62.86%, and that for the control group was 36.00% and 27.78% respectively. Ridit test showed significant difference among the effectiveness in the three groups, while chi 2 showed the difference of the total effective rate in the three groups was insignificant, but that of the markedly effective rate was significant. Serum IL-8 and plasma endothelin levels of 11 cases in the treated group were examined before and after treatment, the results showed that both the two parameters were reduced after the combination therapy.. The effect of combination therapy of CSDD and DYT was better than that of CSDD or DYT alone, therefore, it is worth further studying and spreading.

Topics: Adolescent; Adult; Aged; Drug Therapy, Combination; Drugs, Chinese Herbal; Endothelins; Female; Follow-Up Studies; Humans; Interleukin-8; Male; Middle Aged; Phytotherapy; Psoriasis

Psoriasis is a chronic inflammatory skin disease in which epidermal hyperplasia results from the release of cytokines by infiltrating type 1 T cells. Up- regulation of endogenous interleukin-10 controls type 1 skin responses in animal models; however, interleukin-10 production is low in psoriatic lesions. Consistent with an important role of interleukin-10 in psoriasis, we and colleagues have recently demonstrated clinical efficacy of subcutaneous administration of recombinant interleukin-10 to affected patients. Here, we studied the effects of interleukin-10 on disease-related inflammatory pathways. Patients were treated with recombinant interleukin-10 over 6 wk in an open-label phase II clinical trial. Tissue was obtained before and after therapy and examined by histology/immunohistochemistry, in situ hybridization, and quantitative real-time reverse transcription-polymerase chain reaction. Ten of 14 patients showed a marked reduction of the clinical disease activity. The clinical response was associated with a significant decrease of cutaneous T cell infiltration and the lesional expression of type 1 cytokines interferon-gamma and tumor necrosis factor-alpha. Interleukin-10 inhibited the epidermal interleukin-8 pathway by downregulating the expression of interleukin-8, its receptor CXCR2, and its inducer interleukin-17, and partially reversed the aberrant keratinocyte maturation defining psoriatic epidermal pathology. Remarkably, there was evidence that genetic factors are involved in the response to interleukin-10 as individual variations in the downregulation of tumor necrosis factor-alpha were related to the presence of polymorphisms in the tumor necrosis factor-alpha promoter. These data suggest that excessive production of type 1 cytokines in human skin disease can be counter-regulated by the administration of recombinant interleukin-10. Genotypic analysis may help to identify patients that will preferentially respond to interleukin-10 therapy.

Topics: Cell Differentiation; Cell Division; Cytokines; Dermatitis; Down-Regulation; Epidermis; Female; Humans; Interleukin-10; Interleukin-8; Keratinocytes; Male; Polymorphism, Genetic; Promoter Regions, Genetic; Psoriasis; Receptors, Interleukin-8B; Signal Transduction; Skin; T-Lymphocytes; Tumor Necrosis Factor-alpha

Topics: Adult; Aged; Anti-Bacterial Agents; Cells, Cultured; Clarithromycin; Cytokines; Erythromycin; Female; Humans; Interleukin-8; Keratinocytes; Male; Middle Aged; Psoriasis; Treatment Outcome

Topics: Adult; Chronic Disease; Disease Progression; Humans; Interleukin-8; Psoriasis; Recurrence

Calcipotriene is a synthetic analogue of 1,25-dihydroxyvitamin D3 established to be effective topically in the treatment of psoriasis. We investigated the early cellular and immunological events induced by calcipotriene in psoriasis. Thirty patients with moderate plaque-type psoriasis were randomly assigned to receive twice daily applications of either calcipotriene ointment 0.005% or matching vehicle for 6 weeks. Skin biopsies (6 mm) were performed from designated plaques at baseline and days 3 and 7. On these days and at weeks 2, 4 and 6, complete clinical evaluations were made in a double-blind fashion. Consistent with previous studies, significant clinical improvement (P < 0.05) in psoriasis was observed in patients receiving calcipotriene vs. those receiving vehicle by day 7 for scale and erythema, and by day 14 for thickness. No significant improvement, however, was seen on day 3. None of the immunohistological markers (CD1a, CD4, CD8, ICAM-1, VCAM-1, E-selectin, HLA-DR) semiquantitatively assessed in psoriatic plaques was significantly changed by calcipotriene treatment for 7 days. In the calcipotriene-treated group, interleukin (IL)-10 levels (pg/microgram of protein) increased by 57% from baseline (0.030 +/- 0.006; mean +/- SEM) to day 3 (0.047 +/- 0.011) (P = 0.05 vs. baseline; n = 10) and remained elevated at day 7 (0.046 +/- 0.012). IL-8 levels (pg/microgram of protein), however, declined by 70% from baseline (0.13 +/- 0.06) to day 3 (0.04 +/- 0.01), and remained low at day 7 (0.03 +/- 0.02) (P < 0.05 vs. baseline; n = 10). Both IL-8 and IL-10 were unaffected by vehicle treatment. Calcipotriene-induced clinical improvement of psoriasis is preceded by an increase in IL-10 and a concomitant decrease in IL-8 levels. The changes in the level of these two cytokines provide further evidence for immunological changes as a significant part of the mechanism of action of calcipotriene in psoriasis.

Topics: Adolescent; Adult; Aged; Calcitriol; Dermatologic Agents; Double-Blind Method; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-10; Interleukin-8; Male; Middle Aged; Psoriasis; Treatment Outcome

Recently, the keratinocyte IL-8/IL-8 receptor (IL-8R) pathway has been implicated in the pathogenesis of psoriasis, and there is evidence that the potent macrolide immune suppressant tacrolimus (formerly FK506) can inhibit this pathway in vitro. In this study, determination of the expression of cytokine mRNAs in lesional skin of patients with active disease by reverse transcriptase polymerase chain reaction revealed transcripts for IL-1 beta, tumour necrosis factor-alpha (TNF-alpha), IL-6, IL-8, IL-8R, IL-10, interferon-gamma (IFN-gamma), IL-2R and transforming growth factor-beta (TGF-beta), but not IL-2 or IL-4. IL-8 was the only cytokine expressed in affected skin of all patients but not in clinically normal skin of healthy subjects. In seven CD4+ T cell clones propagated from the lesional skin of an untreated psoriasis patient, IL-8 was expressed by the skin-derived T lymphocytes and not by feeder cells (irradiated autologous blood lymphocytes); IL-1 beta, IL-2, IL-6 and IL-10 were also expressed by some or all of the T cell clones. IL-8 mRNA was not detected in the skin of any patient after the start of systemic tacrolimus therapy; IL-1 beta, IL-6 and IFN-gamma transcripts were also reduced. By 12 weeks, the mean psoriasis area and severity index (PASI) had decreased from 18.8 to 3.8, a reduction of 80%. In the same post-treatment biopsies, however, message for IL-8R persisted. Estimation of circulating IL-8 levels by enzyme immunoassay showed that all patients with detectable IL-8 before treatment had decreased levels in response to treatment with tacrolimus; reductions in PASI scores were accompanied by decreases in IL-8 levels, that varied both in rate and extent. Partial relapse, which in a minority of patients followed the initial period of remission, and was precipitated by drug dose reduction, was accompanied by an increase in circulating IL-8. These findings add credence to the view that the IL-8/IL-8R autocrine/paracrine pathway may be important in the pathogenesis of psoriasis. They further suggest that interference with IL-8 production and/or that of other key chemokines may be an important mechanism underlying the therapeutic efficacy of tacrolimus, and other agents such as cyclosporin A, with similar molecular actions.

Topics: Adolescent; Adult; Cytokines; Female; Gene Expression; Humans; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Psoriasis; Receptors, Interleukin; Receptors, Interleukin-8A; RNA, Messenger; T-Lymphocytes; Tacrolimus

An occlusive dressing that is both cosmetically acceptable and long term is needed for psoriasis treatment. The mechanisms that underlie the efficacy of occlusion in psoriasis are unknown.. We performed a clinical and immunohistologic study in patients with psoriasis of the effects of occlusion, topical corticosteroid alone, and occlusion plus corticosteroid, with a new prolonged dressing as the occlusive therapy.. Nineteen patients completed a 3-week study of efficacy of prolonged occlusion dressing, fluocinonide ointment, or a combination of the two. An immunohistologic study was performed in 10 patients with psoriasis treated for 1 week with prolonged occlusion.. The combination of fluocinonide ointment and occlusion produced significantly more improvement than either treatment alone (p < 0.01). There was no significant difference between the efficacy of prolonged occlusion or fluocinonide ointment. On 4-week follow-up plaques treated with occlusion alone or combined fluocinonide and occlusion were still significantly improved (p = 0.05 and p < 0.001, respectively). None of the immunohistologic and proliferation markers assessed in psoriatic plaques was significantly affected by occlusion as compared with untreated plaques.. Prolonged occlusion is an effective therapy for psoriasis either as monotherapy or in combination with a high-potency topical corticosteroid. However, the mechanism of action of prolonged occlusion alone in the improvement of psoriasis is unknown.

Topics: Administration, Cutaneous; Adult; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Adhesion Molecules; Combined Modality Therapy; E-Selectin; Female; Fluocinonide; Follow-Up Studies; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Langerhans Cells; Male; Middle Aged; Occlusive Dressings; Psoriasis; Receptors, Immunologic

Moxibustion is a traditional medical intervention of traditional Chinese medicine. It refers to the direct or indirect application of ignited moxa wool made of mugwort leaves to acupuncture points or other specific parts of the body for either treating or preventing diseases. Moxibustion has been proven to be effective in treating skin lesions of psoriasis.. This study was performed to elucidate molecular mechanisms underlying the effects of moxibustion treatment on imiquimod-induced psoriatic mice.. We established an imiquimod (IMQ)-induced psoriatic mice (Model) and assessed the effects of moxibustion (Moxi) treatment on skin lesions of psoriatic mice by the PASI scores and expressions of inflammation-related factors relative to normal control mice (NC). We then performed nuclear magnetic resonance (NMR)-based metabolomic analysis on the skin tissues of the NC, Model and Moxi-treated mice to address metabolic differences among the three groups.. Moxi mice showed reduced PASI scores and decreased expressions of the pro-inflammatory cytokines IL-8, IL-17A and IL-23 relative to Model mice. Compared with the Model group, the NC and Moxi groups shared 9 characteristic metabolites and 4 significantly altered metabolic pathways except for taurine and hypotaurine metabolism uniquely identified in the NC group. To a certain extent, moxibustion treatment improved metabolic disorders of skin lesions of psoriatic mice by decreasing glucose, valine, asparagine, aspartate and alanine-mediated cell proliferation and synthesis of scaffold proteins, alleviating histidine-mediated hyperproliferation of blood vessels, and promoting triacylglycerol decomposition.. This study reveals the molecular mechanisms underlying the effects of moxibustion treatment on the skin lesions of psoriasis, potentially improving the clinical efficacy of moxibustion.

Topics: Alanine; Animals; Asparagine; Aspartic Acid; Cytokines; Disease Models, Animal; Glucose; Histidine; Imiquimod; Interleukin-17; Interleukin-23; Interleukin-8; Magnetic Resonance Spectroscopy; Mice; Moxibustion; Psoriasis; Skin; Taurine; Triglycerides; Valine

Psoriasis, an immune-mediated chronic inflammatory skin condition, is treatable with Qinzhu Liangxue (QZLX), a therapeutic medicinal plant formula used in clinical practice. However, further investigation is needed to clarify its molecular mechanisms of action.. The potential biological mechanisms of QZLX to alleviate psoriasis involving IL-6-induced hyperproliferation and inflammation by regulating METTL14/SOCS3/STAT3 axis.. HaCaT cell model was induced by IL-6, and dealt with serum containing QZLX. In addition, shRNAs and siRNAs were used for gene silencing, viruses were collected 48 h post-transfection and infected HaCaT cells. Cell viability was detected by CCK-8 assay, cell cycle was determined by flow cytometry. Finally, psoriasis mice model was induced by IMQ cream, then back skin tissue was used for hematoxylin and eosin (H&E). The content of IL-1β, IL-6, and IL-8 in cell supernatants were analyzed using ELISA kits. Analysis of SOCS3 was used by quantitative RT-PCR, the expression level of SOCS3, METTL3, METTL14, WTAP, SOCS3, YTHDF2, p-STAT3 and STAT3 in HaCaT cells transduced with METTL14 overexpression was detected by Western blot.. All results indicated that QZLX could significantly alleviate IL-6-induced HaCaT cell viability, cell cycle progression, and inhibit the level of IL-1β, IL-6, and IL-8. The m6A levels and level of METTL14 in HaCaT cells treated with IL-6 were enhanced, while it was reversed by QZLX. METTL14 silencing could inhibit IL-6-induced HaCaT cell viability, cell cycle progression and inflammation response, while SOCS3 overexpression also suppressed METTL14-induced HaCaT cell viability, cell cycle progression and inflammation. QZLX could significantly enhance the expression level of SOCS3, while inhibit the level of METTL14, and p-STAT3/STAT3. In addition, QZLX inhibits METTL14-induced HaCaT cell viability, cell cycle progression, and inhibits the level of IL-1β, IL-6, and IL-8.. Our finding suggested that QZLX ameliorated the inflammation response of psoriasis and performed the potential anti-psoriasis effect by regulating METTL14/SOCS3/STAT3 axis in both mice and HaCaT cells psoriasis model. Therefore, our study demonstrated a significant strategy for inhibiting psoriasis inflammation via targeting METTL14/SOCS3/STAT3 axis.

Topics: Animals; Cell Proliferation; HaCaT Cells; Humans; Inflammation; Interleukin-6; Interleukin-8; Keratinocytes; Mice; Psoriasis; STAT3 Transcription Factor

The idea of psoriatic disease continuum has been progressively prompted based on the advances of the knowledge about the pathogenic steps underpinning the occurrence of psoriasis (PSO) and psoriatic arthritis (PSA). To evaluate biomolecules (inflammatory cytokines, inflammatory chemokines, cell adhesion and cellular mediators) in naïve patients with PSO, PSA with PSO, and PSA sine PSO. To stratify the results considering the presence of psoriatic nail involvement, extensive skin disease and obesity evaluating all involved patients.. By multiplex technology, 20 serum biomolecules were assessed with the inclusion of pro-inflammatory cytokines (GM-CSF, IFN-γ, IL-1α, IL-1β, IL-6, IL-8, IL-12p70, IL-17A, IL-23, TNF), anti-inflammatory cytokines (IFN-α, IL-4, IL-10, IL-13), inflammatory chemokines (IP-10, MCP-1, MIP-1α, MIP-1β), cell adhesion and cellular mediators (ICAM-1, E-selectin, P-selectin). The assessment of possible statistical differences between the means of the three groups was performed by One-Way ANOVA. In addition, by non-parametric T-tests, we stratified the results according to selected clinical characteristics (psoriatic nail involvement, PASI ≥ 10, BMI ≥ 30).. In 80 assessed naïve patients, patients with PSO showed significant increases of E-selectin (p=0.021) and IL-8 (0.041) than other groups. In patients with PSA with PSO, significant higher levels of ICAM-1 were observed (p=0.009) than other groups. We did not observe further differences comparing pro-inflammatory and anti-inflammatory cytokines, inflammatory chemokines, and cell adhesion and cellular mediators in patients with PSO, PSA with PSO, and PSA sine PSO. Patients with psoriatic onychopathy showed significant increased levels of ICAM-1 (p=0.010) and IP-10 (0.030) than others. In patients with PASI ≥ 10, significantly enhanced values of IL-8 (p=0.004), TNF (p=0.013), E-selectin (p=0.004), MIP-1α (p=0.003), and MIP-1β (p=0.039). In patients with BMI ≥ 30, significantly higher levels of E-selectin were pointed out (p=0.035) than others.. Our findings may suggest that a similar cytokine profile may characterize naïve patients with PSO, PSA with PSO, and PSA sine PSO, reinforcing the concept of psoriatic disease continuum. However, some differences may be also shown, underlying possible pathogenic differences and leading to the clinical heterogeneity of these patients.

Topics: Arthritis, Psoriatic; Chemokine CCL3; Chemokine CCL4; Chemokine CXCL10; Cytokines; E-Selectin; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Psoriasis

Topics: Coenzyme A Ligases; Cytokines; Ferroptosis; Humans; Interleukin-6; Interleukin-8; Psoriasis; Reactive Oxygen Species; Tumor Necrosis Factor-alpha

Cytokines like IL-17A have been consistently found to be elevated in psoriatic lesional skin, and therapeutic antibodies to IL-17 have demonstrated efficacy in treating psoriatic skin and joint disease. However, results about the circulating cytokines in psoriasis patients remained controversial. Anticytokine autoantibodies (ACAAs) were detected in various autoimmune diseases but remained largely unknown in psoriasis. We aimed to investigate the serum levels of cytokines and ACAAs in psoriasis patients. The study included 44 biologics-naive psoriasis patients and 40 healthy controls. Serum cytokines and the corresponding autoantibodies were measured by multiplex bead-based technology. The bioactivity of serum IL-17A was determined by IL-8 production in primary keratinocytes. Herein, we found serum levels of IL-12B (median: 6.16 vs. 9.03,

Topics: Antibodies, Anti-Idiotypic; Antibodies, Monoclonal, Humanized; Autoantibodies; Biological Products; Cytokines; Humans; Interleukin-17; Interleukin-8; Psoriasis

Skin surface proteins are potential biomarkers in psoriasis and can be measured noninvasively with the transdermal analysis patch (TAP). This study aimed to assess markers measured by TAP over time in daily clinical practice, explore their correlation with disease severity in pediatric psoriasis, and compare the TAP and tape stripping detection capability.. In this prospective observational daily clinical practice study, pediatric psoriasis patients (aged >5 to <18 years) were followed during 1 year. At each visit, TAPs were applied to lesional (n = 2), peri-lesional (n = 2), and non-lesional (n = 1) sites. Post-lesional skin was sampled if all lesions on the arms, legs, or trunk cleared. Treatment and psoriasis severity data were collected. IL-1RA, hBD-2, IL-1α, IL-8, VEGF, CXCL-1/2, CCL-27, IL-23, hBD-1, IL-22, IL-17A, KLK-5, and IL-4 levels were quantified by spot-ELISA. For the statistical analysis, Wilcoxon signed rank tests, Mann-Whitney U tests, and Spearman correlations were used. Detection capability of the TAP was compared to tape stripping in a separate cohort of adult psoriasis patients.. 32 patients (median age 15.0 years, median Psoriasis Area and Severity Index [PASI] 5.2) were followed for a mean of 11.3 (±3.4) months with a total of 104 visits. In lesional skin (n = 197), significantly higher IL-1RA, hBD-2, IL-8, VEGF, CXCL-1/2, IL-23, hBD-1, IL-22, CCL-27, and IL-17A levels were found compared to non-lesional skin (n = 104), while IL-1α was higher in non-lesional skin. Marker levels were highly variable over time and did not correlate with disease severity measured by PASI or SUM scores. Comparison of the TAP and tape strip detection capability in adult psoriasis patients (n = 10) showed that lesional hBD-2, IL1-α, IL-8, and VEGF and non-lesional IL-1RA, hBD-2, IL-8, and VEGF were more frequently detected in tape extracts than TAPs.. Due to the lack of correlation with clinical disease severity and the current detection capability of the markers measured by TAP in psoriasis, its use in regular practice is still a bridge too far.

Topics: Adolescent; Adult; Biomarkers; Child; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-17; Interleukin-23; Interleukin-8; Longitudinal Studies; Membrane Proteins; Psoriasis; Skin; Vascular Endothelial Growth Factor A

Few studies have addressed the impact of the psoriasis-related proinflammatory cytokines on the proliferation and melanogenesis of melanocytes (MCs) in lesional psoriatic skin.. We investigated the effects of TNFα, IL17A, and IL8 on the proliferation and melanin synthesis of MCs.. Skin specimens were biopsied from patients with psoriasis vulgaris at the active stage, or from the tail skin of Dct-LacZ mice with imiquimod (IMQ)-induced psoriasiform dermatitis. Cultured keratinocytes (KCs), MCs, and human skin explants were used in this study. The numbers of MCs were measured via β-galactosidase staining, EdU incorporation and HMB45 immunohistochemical staining. The expression of human β-defensin 3 (hBD3) in KCs was silenced by siRNA, the conditioned medium (CM) from siRNA-transfected KCs was used to treat MCs, then followed by αMSH stimulation. The melanogenesis-related genes were examined by using qRT-PCR and western blotting.. The increased number of MCs and decreased melanin content were highly relevant to the enhanced expression of IL8 and BD3 both in human psoriatic skin and in IMQ-treated mouse tail skin. IL8 expression in KCs and CXCR2 expression in MCs was significantly increased by IL17A and TNFα, the αMSH-induced upregulations of microphthalmia-associated transcription factor (MITF) and tyrosinase in MCs were abrogated by the CM from hBD3-unsilenced KCs, but not from hBD3-silenced KCs.. Our results suggest the roles of IL8-CXCR2 activation in promoting MC proliferation and of BD3 upregulation in reducing melanogenesis. These findings have been implicated in the underlying mechanism that active psoriasis prefers hypopigmentation despite chronic inflammation.

Topics: Animals; Cell Proliferation; Cytokines; Humans; Imiquimod; Interleukin-8; Melanins; Melanocytes; Mice; Microphthalmia-Associated Transcription Factor; Psoriasis; RNA, Small Interfering; Tumor Necrosis Factor-alpha

Cutaneous lichen planus (CLP) and psoriasis (PSO) are both common chronic inflammatory skin diseases for which development of new treatments requires the identification of key targets. While PSO is a typical Th17/IL-17-disorder, there is some evidence that Th1/IFN-ɣ dominate the inflammatory process in CLP. Nonetheless, the immunopathogenesis of CLP is not fully explained and key immunological factors still have to be recognized. In this study, we compared the immune signature of CLP lesions with the well-characterized inflammation present in PSO skin. First, we analysed the histological and immunohistological characteristics of CLP and PSO. Second, we assessed the cytokine expression (IL1A, IL1B, IL4, IL6, IL8, IL10, IL17A, IL19, IL21, IL22, IL23A, IL13, IFNG, TNF, IL12A, IL12B and IL36G) of lesional skin of CLP with PSO by qPCR. Histology revealed a similar epidermal thickness in CLP and PSO. Immunohistochemically, both diseases presented with an inflammatory infiltrate mainly composed by CD3

Topics: Adolescent; Adult; Aged; CD4-Positive T-Lymphocytes; Child; Cytokines; Female; Gene Expression; Humans; Immunohistochemistry; Interferon-gamma; Interleukin-17; Interleukin-8; Interleukins; Janus Kinase 1; Lichen Planus; Male; Middle Aged; Psoriasis; RNA, Messenger; STAT1 Transcription Factor; Young Adult

Psoriasis is a chronic inflammatory skin disease with numerous involved factors. miR-146a and miR-146b (miR-146a/b) are anti-inflammatory miRNAs that are increased in psoriatic skin. SERPINB2 has been shown to be upregulated in the inflammation and infections. Here we aimed to study the relationship between miR-146a/b and SERPINB2 and to delineate the role of SERPINB2 in association of plaque psoriasis. We report increased SERPINB2 expression in the skin of psoriasis patients, which was in a positive relationship with psoriasis severity and in a negative relationship with miR-146a/b in psoriatic lesions. In cultured keratinocytes, both cellular and secreted SERPINB2 levels were strongly induced in response to IFN-γ and TNF-α. Interestingly, SERPINB2 mRNA was downregulated by IL-17A and the combination of TNF-α and IL-17A at time points when miR-146a was increased. The predicted binding site for miR-146a/b in 3' untranslated region of SERPINB2 revealed no activity in luciferase assay, while siRNA silencing of miR-146a/b direct targets IRAK1 and CARD10 resulted in reduced expression of SERPINB2, suggesting that miR-146a/b indirectly control SERPINB2 expression in the skin. The siRNA silencing of SERPINB2 increased the expression of IL-8, CXCL5 and CCL5 and migration of neutrophils revealing its anti-inflammatory role in keratinocytes. Our data together suggest that SERPINB2 and miR-146a/b are part of disease-related network of molecules that are coordinately regulated and act in controlling the inflammatory responses in psoriatic skin.

Topics: CARD Signaling Adaptor Proteins; Case-Control Studies; Cell Movement; Cells, Cultured; Chemokine CCL5; Chemokine CXCL5; Down-Regulation; Gene Silencing; Humans; Inflammation; Interferon-gamma; Interleukin-1 Receptor-Associated Kinases; Interleukin-17; Interleukin-8; Keratinocytes; MicroRNAs; Neutrophils; Psoriasis; RNA, Messenger; RNA, Small Interfering; Severity of Illness Index; Tumor Necrosis Factor-alpha; Up-Regulation

Topics: Caveolin 1; Cells, Cultured; Humans; Inflammation; Interleukin-6; Interleukin-8; Keratinocytes; Leptin; Obesity; Psoriasis

Psoriasis is a recrudescent chronic immune-mediated inflammatory dermatosis; the production and release of proinflammatory cytokines/chemokines such as TNF-α has been regarded as critical issues during psoriasis pathogenesis. Based on online microarray profiles, the expression of the transcription factor GATA3 was downregulated in psoriasis lesion tissues. In the present study, we searched for miRNAs that might be related to TNF-α and GATA3 to investigate an in-depth understanding of psoriasis pathogenesis. Herein, higher TNF-α and GATA3 protein levels were observed in psoriasis lesion tissues and that GATA3 overexpression significantly reverses TNF-α-induced increases within the production of IL-6 and CXCL8 in keratinocytes. TNF-α stimulation increases miR-155 expression dose-independently, and the miR-155 inhibitor significantly reverses TNF-α-induced suppression of GATA3 protein levels and increases IL-6 and CXCL8 production. miR-155 could suppress the expression of GATA3 by targeting its 3'UTR, while GATA3 could activate the transcription of IL37 by targeting its promoter region. miR-155 overexpression reduces IL37 protein and increases CXCL8 production; GATA3 overexpression might significantly attenuate the effects of miR-155 overexpression. In contrast to GATA3, miR-155 expression is significantly upregulated in psoriasis lesion tissue and is negatively correlated with GATA3 and IL37. In summary, the miR-155/GATA3/IL37 axis modulates the production of IL-6 and CXCL8 upon TNF-α stimulation to affect psoriasis development. Thus, miR-155/GATA3/IL37 may be potent targets for psoriasis treatment, which needs further in vivo and clinical investigation.

Topics: 3' Untranslated Regions; GATA3 Transcription Factor; HaCaT Cells; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Keratinocytes; MicroRNAs; Promoter Regions, Genetic; Psoriasis; Skin; Transfection; Tumor Necrosis Factor-alpha

Topics: Anti-Inflammatory Agents; Chemokine CCL20; Drug Discovery; HaCaT Cells; Humans; Interleukin-17; Interleukin-8; Keratinocytes; Ligands; Psoriasis; Receptors, Interleukin-17; Signal Transduction; Small Molecule Libraries; Workflow

Topics: Aminopyridines; Benzamides; Boron Compounds; Bridged Bicyclo Compounds, Heterocyclic; Cell Line; Cyclic AMP; Cyclopropanes; Down-Regulation; Humans; Inflammation Mediators; Interleukin-18; Interleukin-1alpha; Interleukin-8; Keratinocytes; MAP Kinase Signaling System; Phosphodiesterase 4 Inhibitors; Psoriasis; Thalidomide; Tumor Necrosis Factor-alpha

Palmoplantar pustulosis (PPP) is a refractory, nonbacterial impetigo confined to the palms and soles. Its pathogenesis is still obscure, but it may be associated with the large eccrine sweat glands and pores of palmoplantar skin. PPP is considered to be a localized pustular psoriasis. Interleukin (IL)-8, IL-36γ and IL-36Ra play important roles in the pathogenesis of pustular psoriasis, but their role in PPP is unclear.. To evaluate IL-8, IL-36γ and IL-36Ra expression in PPP, and their relationship with acrosyringia and pustule formation.. mRNA expression was quantified in skin samples from patients with PPP (n = 7), patients with psoriasis vulgaris (PSV; n = 8) and healthy controls (HCs) (n = 6) by reverse-transcription-real-time PCR. Protein expression was characterized by immunohistochemistry (PPP, n = 17; PSV, n = 14; HCs, n = 12). Sweat ducts, including acrosyringia, were stained for epithelial membrane antigen (EMA).. IL-8 mRNA and protein were markedly increased in PPP lesions compared with PSV lesions or HC skin. IL-36γ mRNA and protein were significantly more abundant in PPP lesions than in HC skin. IL-36Ra mRNA was significantly overexpressed in PPP lesions compared with HC skin, but there was no difference in IL-36Ra protein between PPP, PSV and HCs. IL-8 was abundantly expressed by neutrophils in PPP pustules, while IL36Ra was localized in the keratinocytes of PPP, PSV and HC skin. IL-36γ and EMA were colocalized in cells surrounding PPP pustules, and IL-36γ was also expressed in sweat duct cells in the dermis.. IL-8, IL-36γ and IL-36Ra are overexpressed in PPP lesions. IL-8, IL-36γ and acrosyringia, rather than IL-36Ra, are associated with pustule formation in PPP.

Topics: Case-Control Studies; Foot; Hand; Humans; Immunohistochemistry; Interleukin-1; Interleukin-8; Interleukins; Polymerase Chain Reaction; Psoriasis; RNA, Messenger; Skin; Skin Diseases, Vesiculobullous

Scratch injury induces Koebner phenomenon in psoriasis. Smoking is also a risk factor for psoriasis. Keratinocytes can produce various psoriasis-related molecules including TNF, IL1 A, IL1B, IL6, IL12B, IL17C, IL23 A, IL36 A, IL36B, IL36 G, CXCL1, CXCL2, CXCL8, CXCL9, CXCL10, CCL20, IFNB, and CAMP. However, the scratch-induced molecular profiling remains elusive.. To profile the induction pattern of above-mentioned psoriasis-related and keratinocyte-derived molecules by scratch injury in the presence or absence of anti-psoriatic drugs or benzo[a]pyrene, a major environmental pollutant of tobacco smoke.. Confluent normal human keratinocytes were scratched and molecules were assayed by qRT-PCR, ELISA and Western blotting with or without drugs and benzo[a]pyrene.. Among the 18 molecules, the scratch injury on a confluent keratinocyte sheet significantly and selectively upregulated the mRNA expression of four cyto/chemokines, CXCL8, CCL20, IL36G, and TNF, in a scratch-line-number-dependent manner under either low- or high-calcium condition. However, significant protein secretion was only demonstrated for CXCL8 and CCL20. The IL36 G protein was not secreted, but its intracellular level was significantly upregulated by scratch injury, whereas neither the secretion nor the intracellular level of TNF protein was affected by scratch injury. Dexamethasone, but not maxacalcitol nor the phosphodiesterase 4 inhibitor apremilast, partially inhibited the CXCL8 and CCL20 secretion. Benzo[a]pyrene significantly and synergistically enhanced the scratch-induced CCL20 secretion that may explain why smoking is a risk factor for psoriasis.. CCL20 and to a less extent CXCL8 may play a key role in triggering the Koebner phenomenon after scratch injury to keratinocytes.

Topics: Chemokine CCL20; Humans; Interleukin-1; Interleukin-8; Keratinocytes; Psoriasis; Skin

IL-22 induces STAT3 phosphorylation and mediates psoriasis-related gene expression. However, the signaling mechanism leading from pSTAT3 to the expression of these genes remains unclear. We focused on Bcl-3, which is induced by STAT3 activation and mediates gene expression. In cultured human epidermal keratinocytes, IL-22 increased Bcl-3, which was translocated to the nucleus with p50 via STAT3 activation. The increases in CXCL8, S100As and human β-defensin 2 mRNA expression caused by IL-22 were abolished by siRNA against Bcl-3. Although CCL20 expression was also augmented by IL-22, the knockdown of Bcl-3 increased its level. Moreover, the combination of IL-22 and IL-17A enhanced Bcl-3 production, IL-22-induced gene expression, and the expression of other psoriasis-related genes, including those encoding IL-17C, IL-19, and IL-36γ. The expression of these genes (except for CCL20) was also suppressed by the knockdown of Bcl-3. Bcl-3 overexpression induced CXCL8 and HBD2 expression but not S100As expression. We also compared Bcl-3 expression between psoriatic skin lesions and normal skin. Immunostaining revealed strong signals for Bcl-3 and p50 in the nucleus of epidermal keratinocytes from psoriatic skin. The IL-22-STAT3-Bcl-3 pathway may be important in the pathogenesis of psoriasis.

Topics: Active Transport, Cell Nucleus; B-Cell Lymphoma 3 Protein; beta-Defensins; Cells, Cultured; Chemokine CCL20; Enzyme Activation; Gene Expression Regulation; Humans; Interleukin-1; Interleukin-17; Interleukin-22; Interleukin-8; Interleukins; Keratinocytes; NF-kappa B p50 Subunit; Phosphorylation; Proto-Oncogene Proteins; Psoriasis; RNA Interference; RNA, Messenger; RNA, Small Interfering; S100 Proteins; Skin; STAT3 Transcription Factor; Transcription Factors

Innate immune processes are central in the development of the chronic inflammatory skin disease psoriasis. Studying stimulation of keratinocytes, monocytes, and dendritic cells by type I interferons or ligation of Toll-like receptors 1/2, 2/6, or 7, but not 7/8, resulted in enhanced surface expression and secretion of CXC chemokine ligand (CXCL) 16. The corresponding CXC chemokine receptor 6 was expressed on neutrophils whose recruitment into skin is important, especially in early psoriatic disease. Using the recently developed technique real-time deformability cytometry demonstrated that CXCL16 and IL-8 decreased the stiffness and enhanced deformation of neutrophils facilitating transmigration through vessel wall. In addition, CXCL16 potently induced migration of neutrophils and enhanced the chemotactic effect of IL-8. The positive feedback loop was supported by IL-8 enhancing CXCL16 production of neutrophils. Blocking of CXCL16 expression by effective treatment of psoriasis patients with tumor necrosis factor-α blockers further supported the pathogenic role of this chemokine. In summary, the data link innate immune stimulation to CXCL16 upregulation and neutrophil infiltration into skin. CXCL16 could therefore represent a potent future target for treatment of psoriasis.

Topics: Adalimumab; Adult; Biopsy; Chemokine CXCL16; Dendritic Cells; Etanercept; Humans; Immunosuppressive Agents; Interleukin-8; Keratinocytes; Middle Aged; Monocytes; Neutrophil Activation; Neutrophil Infiltration; Primary Cell Culture; Psoriasis; Signal Transduction; Skin; Toll-Like Receptors; Tumor Necrosis Factor-alpha; Up-Regulation

Psoriasis is an autoimmune skin disease characterized by keratinocyte hyperproliferation and chronic inflammation. The pathogenesis of psoriasis involves proinflammatory cytokines, such as tumor necrosis factor (TNF), but the mechanism of keratinocyte activation is not well understood. Here, we show that TNF (10 or 50 ng/mL) stimulates a significant (P < .0001) gene expression and secretion of proinflammatory IL-6, CXCL8 and VEGF from both cultured human HaCaT and normal epidermal human keratinocytes (NHEKs). This effect occurs via activation of the mammalian target of rapamycin (mTOR) signalling complex as shown by Western blot analysis and phospho-ELISAs. Pretreatment with the novel natural flavonoid tetramethoxyluteolin (10-100 μmol L

Topics: Cell Survival; Cells, Cultured; Humans; Inflammation; Interleukin-6; Interleukin-8; Keratinocytes; Luteolin; Macrophages; Phosphatidylinositol 3-Kinases; Psoriasis; Recombinant Proteins; TOR Serine-Threonine Kinases; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Biomarkers provide beneficial information to make diagnoses and monitor the progression of many skin diseases. However, biomarkers produced by skin lesion may be too low at concentration to be detected in the systemic circulation.. To address whether intralesional blood (ILB) is advantageous to detect skin-derived biomarkers over circulation blood (CB) of patients with skin diseases.. ILB was collected as overflowing blood when a small incision was made in lesions of patients with mastocytoma and psoriasis. Concentrations of histamine and Human β-Defensin 2 were determined by ELISA. IL-8 was measured using a cytometric beads array (CBA) kit. IL-8 levels in psoriatic lesions were assessed by immunohistochemical staining and quantitative (q) RT-PCR. MicroRNA levels were measured using qRT-PCR.. Plasma histamine levels were increased in ILB of mastocytoma compared with those in CB. Patients with psoriasis showed increased levels of IL-8, β-Defensin 2 in ILB as compared to those in CB. IL-8 levels in ILB correlated with local PASI scores and therefore reversed to those in CB after attenuation of psoriasis with treatment. Furthermore, ILB in psoriasis patients showed increased miR-203, which was highly expressed in psoriatic epidermis.. ILB contains disease-specific biomarkers at higher concentrations than those in CB, and may be useful for diagnosis and monitoring the progression of skin diseases. Thus, this study illustrates the versatility of ILB with an easy accessibility of biomarkers of chemicals, proteins as well as nucleic acids for a myriad of diseases including inflammatory dermatoses and cancers.

Topics: Adult; Aged; beta-Defensins; Biomarkers; Enzyme-Linked Immunosorbent Assay; Female; Histamine; Humans; Interleukin-8; Male; Mastocytoma, Skin; MicroRNAs; Middle Aged; Psoriasis; Skin

Antagonists of IL-17A and its receptor have proven to be highly effective in the treatment of psoriasis. However, many of the underlying molecular mechanisms involved in the pathogenesis of psoriasis are still to be determined. IκBζ (encoded by the NFKBIZ gene) plays a key role in the development of psoriasis by mediating IL-17A- and IL-17F-driven effects. Both IL-17A and IL-17F expression are increased in lesional psoriatic skin. IL-17A/A and IL-17F/F homodimers as well as the IL-17A/F heterodimer signal through the same receptors. The aim of this study was to characterize the role of the IL-17A/F heterodimer in the regulation of NFKBIZ expression and in the regulation of selected psoriasis-associated genes. We demonstrated that IL-17A/F stimulation of human keratinocytes significantly induced NFKBIZ expression. Moreover, silencing IκBζ by siRNA revealed that IκBζ is a key regulator of IL-17A/F-inducible psoriasis-associated genes, including CCL20, DEFB4, IL-8, CHI3L1 and S100A7. In addition, IL-17A/F-induced NFKBIZ expression was mediated by a mechanism involving the p38 MAPK and NF-κB signalling pathways. In conclusion, we present IκBζ as a novel key regulator of IL-17A/F-driven effects in psoriasis. Thus, antagonists to IL-17A/F or IκBζ may present a targeted approach for treating psoriasis.

Topics: Adaptor Proteins, Signal Transducing; beta-Defensins; Cells, Cultured; Chemokine CCL20; Chitinase-3-Like Protein 1; Gene Expression; Gene Silencing; Humans; I-kappa B Proteins; Interleukin-17; Interleukin-8; Keratinocytes; MAP Kinase Signaling System; NF-kappa B; Nuclear Proteins; Psoriasis; S100 Calcium Binding Protein A7; Tumor Necrosis Factor-alpha

Tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) is a multifunctional cytokine that acts through its receptor fibroblast growth factor-inducible 14 (Fn14). Recent studies demonstrated that the TWEAK/Fn14 signals participate in the development of psoriasis. The purpose of this study was to further explore the effect of Fn14 inhibition on experimental psoriasis. Psoriasis-like skin disease was induced in the wild-type and Fn14-knockout BALB/c mice. We found that Fn14 deficiency ameliorates psoriasis-like lesion in this model, accompanied by less inflammatory cell infiltration and proinflammatory cytokine production in lesional skin. The cutaneous expression of TNF receptor type 2 also decreased in the Fn14-deficient mice. Moreover, the topical application of TWEAK exacerbated psoriatic lesion in the wild-type but not in the Fn14-deficient mice. Furthermore, TWEAK promoted the expression of interleukin 8, keratin 17, and epidermal growth factor receptor (EGFR) but inhibited the expression of involucrin in psoriatic keratinocytes in vitro. Interestingly, such effect of TWEAK was abrogated by an EGFR inhibitor (erlotinib). TWEAK also enhances the proliferation and interleukin-6 production of dermal microvascular endothelial cells under psoriatic condition. In conclusion, TWEAK/Fn14 signals contribute to the development of psoriasis, and involves the modulation of resident cells and the transduction of the EGFR pathway. Fn14 inhibition might be a novel therapeutic strategy for patients with psoriasis.

Topics: Animals; Cell Proliferation; Chemokine CCL5; Cytokines; Disease Models, Animal; Erlotinib Hydrochloride; Humans; Interleukin-6; Interleukin-8; Keratin-17; Keratinocytes; Mice; Mice, Inbred BALB C; Mice, Knockout; Psoriasis; RNA Interference; RNA, Small Interfering; Skin; TWEAK Receptor; Up-Regulation

Psoriasis is an inflammatory skin disease with aberrant keratinocyte proliferation, presumably as a result of immune cell activation. Th17 cytokines like IL-17A and IL-22 are critically implicated in epidermal thickening, altered keratinocyte differentiation and production of innate factors such as antimicrobial peptides. Psoriasis treatment options include modern targeted therapies using anti-cytokine antibodies and traditional non-targeted treatments like anthralin (dithranol). While the mode of action of anti-cytokine antibodies is defined, the effects of topical anthralin on psoriatic skin are not fully understood.. This study aims to unravel the direct effects of anthralin on keratinocyte proliferation, differentiation and production of psoriasis-associated factors.. We tested the effects of anthralin on cell proliferation, cytokeratin expression and changes in the expression of antimicrobial peptides using primary keratinocytes and 3D psoriasis tissue models with and without stimulation of the psoriasis-promoting cytokines IL-17A and IL-22. Moreover, we compared the findings derived from monolayer and multilayer cultures to data derived from lesional skin of patients with psoriasis before and under treatment with anthralin.. Our study shows that anthralin directly induces cell apoptosis in vitro in monolayer cultures but not in 3D psoriasis tissue models treated with IL-17A and IL-22. Yet, keratinocyte proliferation as determined by Ki-67 staining is impaired by anthralin in vivo. In lesional skin but not in 3D psoriasis tissue models anthralin rapidly normalizes cytokeratin (CK)16 expression. Furthermore, anthralin directly inhibits DEFB4 expression in vitro and in vivo, while other antimicrobial peptides and cytokines studied like IL-6 and IL-8 are regulated differently in vitro and in vivo.. Our results show that anthralin directly regulates DEFB4A expression. However, its beneficial effects on psoriasis cannot be explained by direct effects on keratinocyte differentiation or cytokine expression.

Topics: Administration, Cutaneous; Anthralin; Apoptosis; beta-Defensins; Biopsy; Cell Differentiation; Cell Proliferation; Cells, Cultured; Dermatologic Agents; Fluorescent Antibody Technique; Humans; Interleukin-17; Interleukin-22; Interleukin-6; Interleukin-8; Interleukins; Keratin-10; Keratin-16; Keratin-5; Keratinocytes; Ki-67 Antigen; Psoriasis; Skin; Tissue Culture Techniques

Topics: Adult; Dermatologic Agents; Female; Humans; Infliximab; Interleukin-8; Male; Middle Aged; Psoriasis; Retrospective Studies; S100 Calcium Binding Protein A7; Skin; Treatment Outcome

Psoriasis is a common chronic inflammatory and immune-mediated skin disease. Antagonists of TNF-α and, recently, IL-17 have proven to be highly effective in the treatment for psoriasis; however, the molecular mechanisms involved in the pathogenesis of psoriasis are poorly understood. Recently, we presented evidence that IκBζ is a key regulator in the development of psoriasis through its role in mediating IL-17A-driven effects. Like IL-17A, IL-17F is produced by a variety of immune cells, and the expression of IL-17F is increased in psoriatic skin. The purpose of this study was to characterize the role of IL-17F in the regulation of IκBζ expression and to investigate whether IL-17F regulates psoriasis-associated genes in human keratinocytes through IκBζ. Here, we demonstrate that IL-17F stimulation induces IκBζ expression at both the mRNA and the protein levels in normal human keratinocytes. Moreover, silencing IκBζ by siRNA revealed that IκBζ is a key regulator of specific IL-17F-inducible psoriasis-associated genes and proteins, including DEFB4/hBD2, S100A7, CCL20, IL-8 and CHI3L1. In addition, IL-17F-induced IκBζ expression is mediated by a mechanism involving the p38 MAPK and NF-κB signalling pathways, as shown by the clear reduction in IL-17F-mediated expression of IκBζ during chemical inhibition of these two signalling pathways. In summary, we present IκBζ as a novel key regulator of IL-17F-driven effects in psoriasis. Thus, antagonists to IκBζ could potentially provide a more targeted approach for treating psoriasis as well as for treating the other inflammatory and immune-mediated diseases for which IL-17-targeting drugs have recently been approved.

Topics: Adaptor Proteins, Signal Transducing; Antibodies, Neutralizing; beta-Defensins; Chemokine CCL20; Chitinase-3-Like Protein 1; Gene Expression; Humans; I-kappa B Proteins; Interleukin-17; Interleukin-8; Keratinocytes; MAP Kinase Signaling System; NF-kappa B; Nuclear Proteins; p38 Mitogen-Activated Protein Kinases; Psoriasis; RNA, Messenger; RNA, Small Interfering; S100 Calcium Binding Protein A7; Transfection; Tumor Necrosis Factor-alpha

Interleukin-36 cytokines are predominantly expressed by epithelial cells. Significant upregulation of epidermal IL-36 is now a recognised characteristic of psoriatic skin inflammation. IL-36 is known to induce inflammatory responses in dendritic cells, fibroblasts and epithelial cells. Although vascular alterations are a hallmark of psoriatic lesions and dermal endothelial cells are well known to play a critical role in skin inflammation, the effects of IL-36 on endothelial cells are unexplored. We here show that endothelial cells including dermal microvascular cells express a functionally active IL-36 receptor. Adhesion molecules VCAM-1 and ICAM-1 are upregulated by IL-36γ stimulation, and this is reversed by the presence of the endogenous IL-36 receptor antagonist. IL-36γ-stimulated endothelial cells secrete the proinflammatory chemokines IL-8, CCL2 and CCL20. Chemotaxis assays showed increased migration of T-cells following IL-36γ stimulation of endothelial cells. These results suggest a role for IL-36γ in the dermal vascular compartment, and it is likely to enhance psoriatic skin inflammation by activating endothelial cells and promoting leucocyte recruitment.

Topics: Chemokine CCL2; Chemokine CCL20; Endothelial Cells; Human Umbilical Vein Endothelial Cells; Humans; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-8; NF-kappa B; Proto-Oncogene Proteins c-jun; Psoriasis; Receptors, Interleukin; T-Lymphocytes; Vascular Cell Adhesion Molecule-1

Interleukin-8 (IL-8) is an important factor in the pathogenesis of psoriasis vulgaris, which is characterized by proliferation of keratinocytes, neutrophil infiltration and angiogenesis. Cysteine-rich 61 (Cyr61/CCN1), a secreted extracellular matrix protein, is a novel proinflammatory factor. Whether Cyr61 is involved in the development of psoriasis vulgaris via IL-8 production remains unknown. In this study we explore the role of Cyr61 in IL-8 expression regulation in vivo and in vitro.. Skin samples from normal donors and psoriasis vulgaris patients were examined the profile of Cyr61 and IL-8 using immunohistochemistry, real-time PCR and Western blotting. HaCaT cells were treated with Cyr61 and IL-8 expression was analyzed by real-time PCR and ELISA. Signal transduction pathways in Cyr61-induced IL-8 production were investigated by real-time PCR, western blotting, luciferase reporter assay or chromatin immunoprecipitation (ChIP) assay. IMQ-induced psoriasis-like mice were treated with anti-Cyr61monoclonal antibodies (mAb), or IgG1 as a control.. We found that Cyr61 was abundant in the epidermis of patients with psoriasis vulgaris and positively correlated with the pathogenesis of skin lesions. Cyr61 induced IL-8 production by keratinocytes in a dose dependent manner. This IL-8 synthesis occurred in an IL-1β- and TNF-α- independent mode via PI3K, AKT and JNK activation, with binding of enhanced AP-1, C/EBPβ and NF-κB to sites located in the IL-8 promoter region. Treatment with anti-Cyr61 mAb led to reduction of MIP-2 level, decreased neutrophil infiltration, and attenuated inflammation in vivo.. Our results not only reveal a novel mechanism illustrating the role of Cyr61 in epidermis pathogenesis but also suggest that therapies targeting Cyr61 may represent a novel strategy in the treatment of psoriasis vulgaris.

Topics: Adult; Animals; Cell Line; Cysteine-Rich Protein 61; Female; Humans; Interleukin-1beta; Interleukin-8; Keratinocytes; Male; MAP Kinase Signaling System; Mice, Inbred BALB C; Middle Aged; NF-kappa B; Psoriasis; RNA, Small Interfering; Skin; Tumor Necrosis Factor-alpha; Young Adult

Psoriasis is a chronic skin disease also affecting other sites such as joints. This disease highly depends on inflammation and angiogenesis as well as other pathways. At each step of the psoriasis molecular pathway, different inflammatory cytokines and angiogenic growth factors are involved such as hypoxia inducible factor-1 α (HIF-1 α), vascular endothelial growth factor (VEGF), matrix metalo proteinases (MMPs), basic fibroblast growth factor (bFGF), Angiopoitin-2, interleukin-8 (IL-8), IL-17, and IL-2. Beside the mentioned growth factors and cytokines, cellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) which play roles in both angiogenesis and inflammation are also involved in the pathogenesis. Cannabinoids are active compounds of Cannabina Sativa inducing their effects through cannabinoid receptors (CBs). JWH-133 is a synthetic cannabinoid with strong anti-angiogenic and anti-inflammatory activities. This agent is able to inhibit HIF-1 α, VEGF, MMPs, bFGF, IL-8, IL-17, and other mentioned cytokines and adhesion molecules both in vivo and in vitro. Altogether, authors suggest using this cannabinoid for treatment of psoriasis due to its potential in suppressing the two main steps of psoriatic pathogenesis. Of course complementary animal studies and human trials are still required.

Topics: Angiogenesis Inhibitors; Animals; Anti-Inflammatory Agents; Cannabinoids; Cell Movement; Cell Proliferation; Humans; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-17; Interleukin-8; Keratinocytes; Models, Theoretical; Neovascularization, Pathologic; Psoriasis; Receptors, Cannabinoid; Vascular Cell Adhesion Molecule-1; Vascular Endothelial Growth Factor A

Psoriasis is a chronic inflammatory skin disease characterized by altered proliferation and differentiation of keratinocytes as well as infiltration of immune cells. Increased expression of Th17 cells and cytokines secreted by them provides evidence for its central role in the pathogenesis of psoriasis. IL-17A, signature cytokine of Th17 cells was found to be highly differentially expressed in psoriatic lesional skin. However, cellular and molecular mechanism by which IL-17A exerts its function on keratinocyte is incompletely understood. To understand IL-17A mediated signal transduction pathways, gene expression profiling was done and differentially expressed genes were analysed by IPA software. Here, we demonstrate that during IL-17A signaling total cholesterol levels were elevated, which in turn resulted in the suppression of genes of cholesterol and fatty acid biosynthesis. We found that accumulation of cholesterol was essential for IL-17A signaling as reduced total cholesterol levels by methyl β cyclodextrin (MBCD), significantly decreased IL-17A induced secretion of CCL20, IL-8 and S100A7 from the keratinocytes. To our knowledge this study for the first time unveils that high level of intracellular cholesterol plays a crucial role in IL-17A signaling in keratinocytes and may explain the strong association between psoriasis and dyslipidemia.

Topics: Adolescent; Adult; Aged; beta-Cyclodextrins; Cell Differentiation; Cell Proliferation; Chemokine CCL20; Child; Cholesterol; Dyslipidemias; Fatty Acids; Female; Gene Expression Profiling; Gene Expression Regulation; Humans; Interleukin-17; Interleukin-8; Keratinocytes; Male; Middle Aged; Psoriasis; S100 Calcium Binding Protein A7; S100 Proteins; Signal Transduction

Tumour necrosis factor (TNF)-α antagonist therapy is currently used for moderate and severe psoriasis. However, this treatment has several drawbacks, including interindividual variability in clinical response and secondary loss of effectiveness.. To evaluate quantitatively the TNF-α-neutralizing activity of the plasma of patients with psoriasis during TNF-α antagonist therapy and to determine poor responders objectively.. We used a human interleukin-8 reporter monocyte cell line, THP-G8, that harbours a stable luciferase orange (SLO) gene under the control of the interleukin-8 promoter. After confirming its dose-dependent response to exogenous TNF-α, we examined the suppressive activity of TNF-α antagonists and of the patients' plasma during TNF-α antagonist therapy on TNF-α-induced SLO luciferase activity (TNF-SLO-LA).. Pretreatment of TNF-α with TNF-α antagonists or with the plasma of patients with psoriasis who achieved 75% improvement in Psoriasis Area and Severity Index (PASI 75) dose dependently suppressed TNF-SLO-LA. There was a significant correlation between change in PASI and percentage suppression (inhibitory rate of a 1 : 2 dilution of patient plasma on TNF-SLO-LA). A percentage suppression of 50·3% has a positive predictive value of 87% of achieving PASI 75, with a sensitivity of 93% and a specificity of 80%.. Therapeutic monitoring of patients with psoriasis during TNF-α antagonist therapy using THP-G8 can provide a useful tool to determine objectively the efficacy of the administered TNF-α antagonists.

Topics: Adalimumab; Adult; Aged; Antibodies; Cell Line; Dermatologic Agents; Drug Monitoring; Female; Humans; Infliximab; Interleukin-8; Male; Middle Aged; Psoriasis; Severity of Illness Index; Treatment Outcome; Tumor Necrosis Factor-alpha; Young Adult

IL-35 downregulates Th17 cell development and suppresses certain types of autoimmune inflammation such as collagen-induced arthritis and experimental autoimmune uveitis. Psoriasis is thought to be initiated by abnormal interactions between cutaneous keratinocytes and systemic immune cells. However, the role of IL-35 in psoriasis remains unclear. In this study, we assessed IL-35 in three well-known psoriasis models: a human keratinocyte cell line (HaCaT), a keratin 14 (K14)-vascular endothelial growth factor A (VEGF-A)-transgenic (Tg) mouse model, and an imiquimod-induced psoriasis mouse model. First, we found that IL-35 suppressed the expression of IL-6, CXCL8, and S100A7, which are highly upregulated by a mixture of five proinflammatory cytokines in HaCaT. Second, a plasmid coding for the human IL-35 sequence coated with cationic liposomes showed potent immunosuppressive effects on K14-VEGF-A-Tg and imiquimod-induced psoriasis mouse models. In the K14-VEGF-A-Tg model, our results showed that several types of proinflammatory cytokines were significantly reduced, whereas IL-10 was remarkably induced by IL-35. Compared with pcDNA3.1, there was a small number of CD4(+)IL-17(+) T cells and a large number of CD4(+)IL-10(+) and CD4(+)CD25(+)Foxp3(+) T cells in the IL-35 group. Most importantly, we found that IL-35 decreased the total number of macrophages and ratio of M1/M2 macrophages, which has not been reported previously. In addition, compared with dexamethasone, IL-35 showed long-term therapeutic efficacy. In summary, our results strongly indicate that IL-35 plays a potent immunosuppressive role in psoriasis. Thus, IL-35 has potential for development as a new therapeutic strategy for patients with chronic psoriasis and other cutaneous inflammatory diseases.

Topics: Animals; Cells, Cultured; Cytokines; Dexamethasone; Humans; Inflammation; Interleukin-6; Interleukin-8; Interleukins; Keratinocytes; Macrophages; Mice; Mice, Inbred BALB C; Psoriasis; S100 Calcium Binding Protein A7; S100 Proteins; Th17 Cells

Psoriasis is a debilitating chronic inflammatory disease. In addition to the characteristic effects on the skin, chronic inflammation associated with the disease is recognized to contribute to cardiovascular, hepatic and renal comorbidities. Immature myeloid regulatory cells, known as myeloid‑derived suppressor cells (MDSCs), have been demonstrated to accumulate in various diseases and chronic inflammatory states, including inflammatory bowel disease and various types of cancer. The results of the present study, obtained using flow cytometry and cell culture analysis of peripheral blood mononuclear cells from psoriasis and healthy patients, revealed that MDSC levels are significantly increased in the blood of patients with psoriasis compared with healthy controls. Furthermore, these cells are capable of producing various molecules, including matrix metalloproteinase‑9 and‑1, interleukin‑8, growth‑related oncogene, and monocyte chemoattractant protein 1. These molecules may recruit additional immune cells involved in the pathogenesis of the disease, and contribute to the chronic inflammatory state in these patients. Therefore, MDSCs, which have various immune regulatory functions, may contribute to the pathogenesis of psoriasis as a systemic inflammatory disease.

Topics: Adult; Chemokine CCL2; Female; Humans; Inflammation; Interleukin-8; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 9; Middle Aged; Myeloid-Derived Suppressor Cells; Psoriasis

Tumour necrosis factor (TNF)-α inhibition is an effective treatment for moderate to severe plaque-type psoriasis. A change in the cytokine expression profile occurs in the skin after 4 days of treatment, preceding any clinical or histological improvements. MicroRNAs (miRNAs) are important post-transcriptional regulators of gene expression, but miRNA expression has never been studied in psoriatic skin during treatment.. To investigate changes in miRNA expression in psoriatic skin during adalimumab treatment and to compare results with changes in miRNA expression in a mouse model of Aldara-induced psoriasis-like skin inflammation.. Punch biopsies were obtained from nonlesional and lesional psoriatic skin during adalimumab treatment. In the mouse model of Aldara-induced skin inflammation, biopsies were obtained from TNF-α knockout (KO), IL-17A KO and wild-type mice. miRNA expression levels were analysed with microarray, reverse transcriptase quantitative polymerase chain reaction and in situ hybridization.. In psoriatic skin, no changes in miRNA expression were seen 4 days after treatment initiation. After 14 days of treatment, the expression of several miRNAs was normalized towards the level seen in nonlesional skin before treatment. miR-23b expression increased after 14 days of treatment and remained high for 84 days, despite unaltered levels at baseline. In the mouse model of Aldara-induced skin inflammation, the level of miR-146a increased, whereas no regulation was seen for miR-203, miR-214-3p, miR-125a, miR-23b or let-7d-5p.. This study demonstrates that the changes seen in the cytokine expression levels after 4 days of treatment with adalimumab are not facilitated by early changes in miRNA expression.

Topics: Adalimumab; Adult; Aged; Aminoquinolines; Animals; Anti-Inflammatory Agents; Case-Control Studies; Down-Regulation; Female; Humans; Imiquimod; Interleukin-8; Irritants; Male; Mice, Inbred C57BL; Mice, Knockout; MicroRNAs; Middle Aged; Psoriasis; RNA, Messenger

Psoriasis is a systemic immune-mediated chronic inflammatory disease. In the skin, the antimicrobial proteins koebnerisin (S100A15) and psoriasin (S100A7) are overexpressed in the epidermis of psoriatic lesions and mediate inflammation as chemoattractants for immune cells. Their role for systemic inflammation in circulating leukocytes is unknown.. The aim of the study was to identify circulating leukocyte populations as a source of koebnerisin and psoriasin. Further, immune-stimulatory effects of these S100A proteins on circulating leukocytes were evaluated and their role as therapeutic response markers in patients with psoriasis was analyzed upon UVB treatment.. The expression and production of koebnerisin and psoriasin by leukocytes were assessed by quantitative real-time PCR (qRT-PCR) and immunoblotting. The S100A protein mediated regulation of proinflammatory cytokines by peripheral blood mononuclear cells (PBMCs) was measured with qRT-PCR and cytometric bead assay.. We identified circulating leukocytes as novel sources of koebnerisin (S100A15) and psoriasin (S100A7). Circulating leukocytes (PBMCs) of patients with psoriasis produced increased levels of koebnerisin and psoriasin compared to healthy individuals. Both S100A proteins further acted as 'alarmins' on PBMC to induce proinflammatory cytokines implicated in the pathogenesis of psoriasis, such as IL-1β, TNF-α, IL-6 and IL-8. Koebnerisin levels were suppressed in PBMC of psoriatic patients when effectively treated with narrow-band UVB.. Data suggest that koebnerisin and psoriasin are systemic pro-inflammatory mediators and koebnerisin acts as a therapeutic response marker in psoriasis.

Topics: Adult; Aged; Cytokines; Female; Humans; Inflammation; Interleukin-1beta; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Male; Middle Aged; Psoriasis; RNA; S100 Calcium Binding Protein A7; S100 Proteins; Tumor Necrosis Factor-alpha; Ultraviolet Therapy; Young Adult

Topics: Case-Control Studies; Fibroblasts; Humans; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Psoriasis; STAT3 Transcription Factor; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Previous research revealed heterogeneity in the perfusion intensity within clinically homogenous-appearing plaques, without differences in erythema. In addition, an increased perfusion was found within the perilesional skin. This raises the question whether the heterogeneity in perfusion found both inside and outside a lesion influences the expression levels of genes and proteins involved in the pathogenesis of psoriasis.. To correlate the perfusion intensity to mRNA and protein expression of genes associated with the pathogenesis of psoriasis and to visualize the dynamics of the perfusion intensity over time using laser Doppler perfusion imaging.. Fourteen patients with plaque psoriasis were included. The superficial microcirculation and clinical local scores (single usability metric, SUM, scores) were analysed in one representative lesion every 2 weeks. After 8 weeks 4 biopsies were taken, one from a highly perfused area (hotspot) and one from a low perfusion area (coldspot) of the lesional skin, one biopsy from the highly perfused perilesional skin and one from the distant uninvolved skin.. Statistically significant differences in mRNA and protein expression, including IL-17 and TBX21/T-Bet, were found between hotspots and coldspots, and between the highly perfused perilesional and the uninvolved skin. Hotspots tend to remain on the same location during 8 weeks of follow-up.. Within homogenous-appearing psoriatic plaques, there are remarkable differences in mRNA and protein levels, which are correlated with the perfusion intensity and can be detected by using laser Doppler perfusion imaging. In addition, differences in mRNA and protein expression between the highly perfused perilesional skin and the uninvolved skin were found, indicating that several biological changes occur well before clinical changes become manifest.

Topics: Adult; Aged; beta-Defensins; CD3 Complex; Elafin; Female; Gene Expression; Humans; Interleukin-17; Interleukin-8; Keratin-16; Male; Microcirculation; Middle Aged; Platelet Endothelial Cell Adhesion Molecule-1; Psoriasis; RNA, Messenger; Skin; T-Box Domain Proteins

IL-37 is a potent inhibitor of innate immunity by shifting the cytokine equilibrium away from excessive inflammation. Psoriasis is thought to be initiated by abnormal interactions between the cutaneous keratinocytes and systemic immune cells, triggering keratinocyte hyperproliferation. In the current study, we assessed IL-37 in two well-known psoriasis models: a human keratinocyte cell line (HaCaT) and the keratin 14 VEGF-A-transgenic mouse model. First, we used the HaCaT cell line, which was transiently transfected with an overexpressing IL-37 vector, and tested the effect of IL-37 on these cells using a mixture of five proinflammatory cytokines. IL-37 was effective in suppressing the production of CXCL8, IL-6, and S100A7, which were highly upregulated by the mixture of five proinflammatory cytokines. Keratin 14 VEGF-A-transgenic mice were treated with plasmid coding human IL-37 sequence-formulated cationic liposomes, and we observed potent immunosuppressive effects over the 18-d period. In this model, we observed reduced systemic IL-10 levels, local IFN-γ gene transcripts, as well as mild mast cell infiltration into the psoriatic lesions of the mice. Immunohistochemical analysis indicated that IL-37 was expressed by effector memory T cells, as well as macrophages, in human psoriatic plaques. In conclusion, our studies strongly indicate that IL-37 plays a potent immunosuppressive role in the pathogenesis of both experimental psoriasis models in vitro and in vivo by downregulating proinflammatory cytokines. Importantly, our findings highlight new therapeutic strategies that can be designed to use this immunosuppressive anti-inflammatory cytokine in psoriasis and other inflammatory cutaneous diseases.

Topics: Animals; Cell Line; Cell Proliferation; Disease Models, Animal; Down-Regulation; Humans; Immunologic Memory; Immunosuppression Therapy; Inflammation; Interferon-gamma; Interleukin-1; Interleukin-10; Interleukin-6; Interleukin-8; Keratin-14; Keratinocytes; Macrophages; Mast Cells; Mice; Mice, Transgenic; Psoriasis; S100 Calcium Binding Protein A7; S100 Proteins; Skin; T-Lymphocytes; Transfection; Vascular Endothelial Growth Factor A

Topical immunosuppressant therapy is widely used in the treatment of inflammatory skin diseases, such as atopic dermatitis and psoriasis. Besides its beneficial therapeutic effects, application of topical anti-inflammatory drugs may render the epidermis more vulnerable to invading pathogens by suppressing innate immune responses in keratinocytes (KCs). Cytokines, chemokines and antimicrobial peptides (AMPs) produced by epithelial cells enable them to participate in innate and acquired immune responses. The aim of the present work was to study the influence of tacrolimus (FK506) on KCs infected with Malassezia furfur (M. furfur), evaluating the expression of pro-inflammatory cytokines IL-1α and IL-6, chemokine IL-8, anti-inflammatory cytokines transforming growth factor beta1 (TGF-β1) and IL-10 and AMP β-defensin-2. Human KCs were obtained from surgical specimens of normal adult skin. The expression of mRNAs in KCs: FK506-treated, FK506-treated and M. furfur-infected as well as only M. furfur-infected was quantified by real-time quantitative polymerase chain reaction. Next, the production of the AMP β-defensin-2 and of the above-mentioned pro-inflammatory and anti-inflammatory cytokines was evaluated using enzyme-linked immunosorbent assay. In this study, FK506 did not alter cytokine and AMP production by KCs; this led us to hypothesise that it may not enhance the risk of mycotic skin infections.

Topics: Antimicrobial Cationic Peptides; beta-Defensins; Cell Survival; Cytokines; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-10; Interleukin-1alpha; Interleukin-6; Interleukin-8; Keratinocytes; Malassezia; Psoriasis; RNA, Messenger; Skin; Tacrolimus; Transforming Growth Factor beta1

Psoriasis (Ps) is an autoimmune disease characterized by keratinocyte hyperproliferation and chronic inflammation, with increased expression of tumor necrosis factor (TNF) and vascular endothelial growth factor (VEGF). Anti-TNF biologic agents are effective in treating Ps, but are associated with increased risk of infections and blood malignancies. Moreover, keratinocyte hyperproliferation and activation have yet to be addressed. Flavonoids, such as luteolin, are natural compounds with potent anti-inflammatory properties, but their actions on keratinocytes remain unknown. We show that TNF (50 ng/mL) triggers significant production of inflammatory mediators interleukin-6, interleukin-8 and VEGF from both human HaCaT and primary keratinocytes. Pretreatment with the flavonoid luteolin (10-100 µM) significantly inhibits mRNA expression and release of all three mediators in a concentration-dependent manner. More importantly, luteolin decreases TNF-induced phosphorylation, nuclear translocation and DNA binding of the nuclear factor-kappa B (NF-κB) typically involved in inflammatory mediator transcription. We also report that luteolin reduces TNF-induced mRNA expression of two genes (NFKB1 and RELA) encoding two NF-κB subunits (NF-κB p50 and NF-κB p65, respectively). Interestingly, we show that gene expression of RELA is increased in human psoriatic skin. Keratinocyte proliferation, which is a characteristic feature of psoriatic skin, is effectively reduced by luteolin in HaCaT cells, but not in primary keratinocytes. Finally, luteolin does not affect intracellular ATP production or viability. Appropriate formulations of luteolin and related flavones may be promising candidates to be developed into local and systemic treatments for Ps and other inflammatory skin diseases.

Topics: Active Transport, Cell Nucleus; Adenosine Triphosphate; Adult; Blotting, Western; Cell Line; Cell Nucleus; Cell Proliferation; Cells, Cultured; Gene Expression; Humans; Interleukin-6; Interleukin-8; Keratinocytes; Luteolin; NF-kappa B; Phosphorylation; Psoriasis; Reverse Transcriptase Polymerase Chain Reaction; Skin; Transcription Factor RelA; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

"Pustulosis palmaris et plantaris", or palmoplantar pustulosis (PPP), is a chronic pustular dermatitis characterized by intraepidermal palmoplantar pustules. Although early stage vesicles (preceding the pustular phase) formed in the acrosyringium contain the antimicrobial peptides cathelicidin (hCAP-18/LL-37) and dermcidin, the details of hCAP-18/LL-37 expression in such vesicles remain unclear. The principal aim of the present study was to clarify the manner of hCAP-18/LL-37 expression in PPP vesicles and to determine whether this material contributed to subsequent inflammation of lesional skin. PPP vesicle fluid (PPP-VF) induced the expression of mRNAs encoding IL-17C, IL-8, IL-1α, and IL-1β in living skin equivalents, but the level of only IL-8 mRNA decreased significantly upon stimulation of PPP vesicle with depletion of endogenous hCAP-18/LL-37 by affinity chromatography (dep-PPP-VF). Semi-quantitative dot-blot analysis revealed higher concentrations of hCAP-18/LL-37 in PPP-VF compared to healthy sweat (2.87±0.93 µM vs. 0.09±0.09 µM). This concentration of hCAP-18/LL-37 in PPP-VF could upregulate expression of IL-17C, IL-8, IL-1α, and IL-1β at both the mRNA and protein levels. Recombinant hCAP-18 was incubated with dep-PPP-VF. Proteinase 3, which converts hCAP-18 to the active form (LL-37), was present in PPP-VF. Histopathological and immunohistochemical examination revealed that early stage vesicles contained many mononuclear cells but no polymorphonuclear cells, and the mononuclear cells were CD68-positive. The epidermis surrounding the vesicle expresses monocyte chemotactic chemokine, CCL2. In conclusion, PPP-VF contains the proteinase required for LL-37 processing and also may directly upregulate IL-8 in lesional keratinocytes, in turn contributing to the subsequent inflammation of PPP lesional skin.

Topics: Adult; Aged; Aged, 80 and over; Antimicrobial Cationic Peptides; Cathelicidins; Female; Humans; Inflammation; Interleukin-8; Keratinocytes; Male; Middle Aged; Psoriasis; RNA, Messenger; Skin; Sweat Glands

Mutations in the caspase recruitment domain, family member 14 (CARD14) gene have recently been described in psoriasis patients, and explain the psoriasis susceptibility locus 2 (PSORS2). CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling. CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells. In this manuscript, the identity of the dermal cell types expressing CARD14, as well the potential functional consequence of overactive CARD14 in these dermal cell types, was determined. Using two-color immunofluorescence, dermal CARD14 did not co-localize with T-cells, dendritic cells, or macrophages. However, dermal CARD14 did highly co-localize with CD31(+) endothelial cells (ECs). CARD14 was also expressed non-dermal endothelial cells, such as aortic endothelial cells, which may indicate a role of CARD14(+)ECs in the systemic inflammation and cardiovascular comorbidities associated with psoriasis. Additionally, phosphorylated NF-κB was found in psoriatic CARD14(+) CD31(+) ECs, demonstrating this pathway is active in dermal ECs in psoriasis. Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2. These results provide preliminary evidence that CARD14 expression in ECs may contribute to psoriasis through increased expression of chemokines and facilitating recruitment of immune cells into skin.

Topics: CARD Signaling Adaptor Proteins; Cells, Cultured; Chemokine CCL2; Chemokine CXCL10; Dermis; Endothelial Cells; Guanylate Cyclase; Humans; Interleukin-8; Keratinocytes; Membrane Proteins; Monocytes; Mutation; NF-kappa B; Phosphorylation; Platelet Endothelial Cell Adhesion Molecule-1; Psoriasis; RNA, Messenger; Signal Transduction; T-Lymphocytes; Transcriptome; Transfection

Epidermal keratinocytes produce proinflammatory cytokines/chemokines upon stimulation with cytokine milieus and Toll-like receptor ligands, which are considered to reflect epidermal environments in inflamed skin. The human antimicrobial peptide LL-37, besides having microbicidal functions, plays multiple roles as a "host defense peptide" in the immune system. Here, we examined the effect of LL-37 on proinflammatory responses induced by double-stranded RNA (dsRNA) and cytokines in primary human keratinocytes. LL-37 inhibited dsRNA-induced production of thymic stromal lymphopoietin (TSLP), CCL5/RANTES, CXCL10/IP-10, and CXCL8/IL-8, which was attributable to interaction between LL-37 and dsRNA, although LL-37 upregulated CXCL8 expression at an earlier time point (8 h). LL-37 inhibited the increase of CXCL10 and CCL5 induced by TNF-α- and/or IFN-γ but enhanced that of CXCL8. LL-37 and Th17 cytokines (IL-17 and IL-22) synergistically upregulated the expression of CXCL8 and IL-6. LL-37 showed the effects above at a high concentration (25 μg/ml, 5.6 μM). We also examined effects of a peptide with a scrambled LL-37 sequence, which has been frequently used as a negative control, and those of another peptide with the reversed LL-37 sequence, activities of which have not been well investigated. Interestingly, the reversed LL-37 had effects similar to LL-37 but the scrambled LL-37 did not. The modulation by LL-37 of the keratinocyte proinflammatory responses induced by cytokine milieus and dsRNA suggests novel roles for LL-37 in skin inflammation such as the promotion of IL17/IL-22/IL-6-associated psoriasis and suppression of TSLP-associated atopic dermatitis.

Topics: Amino Acid Sequence; Anti-Inflammatory Agents; Antimicrobial Cationic Peptides; Cathelicidins; Cells, Cultured; Chemokine CCL5; Chemokine CXCL10; Cytokines; Dermatitis, Atopic; Enzyme-Linked Immunosorbent Assay; Humans; Interferon-gamma; Interleukin-22; Interleukin-6; Interleukin-8; Interleukins; Keratinocytes; Molecular Sequence Data; Primary Cell Culture; Psoriasis; Recombinant Proteins; RNA, Double-Stranded; Th17 Cells; Thymic Stromal Lymphopoietin; Time Factors; Tumor Necrosis Factor-alpha

Inflammation-associated pigmentation changes are extremely common, but the etiology underlying this clinical observation remains elusive. Particularly, it is unclear how the myriad of cytokines known to be involved in inflammatory skin processes affect epidermal melanocytes. We sought to determine how IL-17 and tumor necrosis factor (TNF) influence normal human melanocytes, as these two cytokines have been implicated in various skin diseases. IL-17 and TNF jointly stimulated broad inductions of cytokines, including melanoma mitogens CXCL1 and IL-8. Moreover, IL-17 and TNF synergistically inhibited pigmentation-related signaling and melanin production, and induced keratinocyte production of β-defensin 3, an antagonist for melanocortin 1 receptor. When analyzing psoriasis lesions that are known to overexpress IL-17 and TNF, we observed an increase in melanocyte number and a simultaneous decrease in pigmentation signaling. Furthermore, therapeutic neutralization of TNF and IL-17 with mAbs resulted in a rapid recovery of pigment gene expression in psoriasis lesions. These results demonstrate that IL-17 and TNF can affect both the growth and pigment production of melanocytes, which may contribute to the pigmentation changes associated with psoriasis. These findings may allow the development of novel therapeutics for pigmentary disorders and bring new insights into the immune milieu surrounding melanocytes and related neoplasms.

Topics: beta-Defensins; Chemokine CXCL1; Cytokines; Epidermis; Gene Expression Profiling; Gene Expression Regulation; Humans; Inflammation; Interleukin-17; Interleukin-8; Melanocytes; Oligonucleotide Array Sequence Analysis; Psoriasis; Real-Time Polymerase Chain Reaction; Signal Transduction; Skin; Tumor Necrosis Factor-alpha

Peptide T (PT), an octapeptide fragment located in the V2 region of the HIV-1 gp120-coating protein, appears to be beneficial in the treatment of psoriasis. Our previous investigations suggest that keratinocytes play a key role in conditioning the therapeutic effects of PT in psoriasis. The aim of this study was to explore the effects of PT and the peptidomimetic natural products, Dhurrin and Prunasin, on the expression of the IL-6, IL-8, IL-23, HSP70 and ICAM-1 on IFN-γ and TNF-α-NHEK activated cells. Moreover, we analysed the interference of PT and its analogues through STAT-3 activation. Our results show that the analogues tested exhibit the beneficial biological effects of PT, suggesting the primary role of keratinocytes upon which PT and the peptidomimetics act directly, by reducing proinflammatory responses. Its reduction appears to be important for therapeutic approach in psoriasis pathogenesis.

Topics: Amygdalin; Antineoplastic Agents, Phytogenic; Enzyme Inhibitors; HSP70 Heat-Shock Proteins; Humans; Inflammation; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-23; Interleukin-6; Interleukin-8; Keratinocytes; Nitriles; Peptide T; Psoriasis; Signal Transduction; STAT3 Transcription Factor; Tumor Necrosis Factor-alpha

Mast cells modulate autoimmune diseases such as psoriasis and multiple sclerosis. Fumaric acid esters (FAEs) are widely used for the treatment of psoriasis, and dimethylfumarate (DMF) has recently been approved for multiple sclerosis. In this study, we analysed the cytotoxic effect of FAEs on human mast cells. Specifically, cell death was analysed in the human mast cell line HMC-1 and in primary cord blood-derived mast cells (CBMCs) after incubation with fumaric acid (FA), monomethylfumarate (MMF), DMF and calcium bis(monomethylfumarate) (Ca-MF). Our data show that only DMF potently induces apoptotic cell death in HMC-1 cells and CBMCs. DMF-mediated apoptosis was associated with increased expression of Bax and Bak and activation of caspase-9 and caspase-6. Interestingly, DMF also enhanced the sensitivity of CBMCs towards TRAIL- and dexamethasone-induced apoptosis. These findings demonstrate for the first time that DMF induces apoptosis of human mast cells, primarily via the mitochondrial apoptotic pathway. Our study contributes to the understanding of the beneficial effects of FAEs in autoimmune diseases and provides a rationale for exploiting FAEs for other diseases associated with mast cells.

Topics: Apoptosis; bcl-2 Homologous Antagonist-Killer Protein; bcl-2-Associated X Protein; Calcium; Caspase 6; Caspase 9; Cell Death; Cell Line; Dermatologic Agents; Dexamethasone; Dimethyl Fumarate; Etoposide; Fumarates; Humans; Interleukin-8; Maleates; Mast Cells; Methotrexate; Psoriasis; TNF-Related Apoptosis-Inducing Ligand

Streptococcal throat infection is associated with a specific variant of psoriasis and with HLA-Cw6 expression. In this study, activation of circulating psoriatic cutaneous lymphocyte-associated antigen (CLA)(+) memory T cells cultured together with epidermal cells occurred only when streptococcal throat extracts were added. This triggered the production of Th1, Th17, and Th22 cytokines, as well as epidermal cell mediators (CXCL8, CXCL9, CXCL10, and CXCL11). Streptococcal extracts (SEs) did not induce any activation with either CLA(-) cells or memory T cells cultured together with epidermal cells from healthy subjects. Intradermal injection of activated culture supernatants into mouse skin induced epidermal hyperplasia. SEs also induced activation when we used epidermal cells from nonlesional skin of psoriatic patients with CLA(+) memory T cells. Significant correlations were found between SE induced upregulation of mRNA expression for ifn-γ, il-17, il-22, ip-10, and serum level of antistreptolysin O in psoriatic patients. This study demonstrates the direct involvement of streptococcal infection in pathological mechanisms of psoriasis, such as IL-17 production and epidermal cell activation.

Topics: Animals; Antigens, Differentiation, T-Lymphocyte; Antistreptolysin; Cells, Cultured; Chemokine CXCL10; Culture Media; Epidermal Cells; Epidermis; Humans; Immunologic Memory; Interferon-gamma; Interleukin-17; Interleukin-22; Interleukin-8; Interleukins; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Pharyngitis; Psoriasis; RNA, Messenger; Streptococcal Infections; Streptococcus; T-Lymphocytes; Tumor Necrosis Factor-alpha

The efficacy of 1α,25-dihydroxyvitamin D3 (Vit-D) limits its topical use despite its profound effects on cellular differentiation, proliferation, and immunomodulation. Therefore, in search for a more effective analog of Vit-D, in this study we have evaluated the antiproliferative and proapoptotic effects of 1α,25-dihydroxyvitamin D3-3-bromoacetate (BE). Proliferation and apoptosis studies in normal human epidermal keratinocytes (NHEKs) were conducted by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), CFSE (carboxy fluorescein succinimidyl ester) dilution, and Annexin V assays. Western blot analysis and real-time PCR were performed to determine its effect on signal transduction. A reconstructed human epidermis (RHE) model was used to further validate the therapeutic role of BE in psoriasis. BE was significantly more potent than an equivalent concentration of Vit-D in inhibiting growth and survival of human keratinocytes. The antimitotic effect was found to be due to the inhibition of phosphorylation of serine/threonine protein kinase (AKT) and its downstream target, mammalian target of rapamycin (mTOR). In the RHE model, BE reversed IL-22-induced psoriasiform changes more effectively than Vit-D. Interestingly, BE inhibited the IL-22-induced gene expression of AKT1, MTOR, chemokines [IL-8 and RANTES (regulated upon activation, normal T-cell expressed and secreted)], and psoriasin (S100A7) more significantly than Vit-D. These results suggest the potential of BE as a prospective therapeutic agent for psoriasis.

Topics: Apoptosis; Calcitriol; Cell Proliferation; Cell Survival; Cells, Cultured; Chemokine CCL5; Epidermal Cells; Gene Expression; Humans; Interleukin-22; Interleukin-8; Interleukins; Keratinocytes; Phosphorylation; Proto-Oncogene Proteins c-akt; Psoriasis; S100 Calcium Binding Protein A7; S100 Proteins; Signal Transduction; TOR Serine-Threonine Kinases; Vitamin D; Vitamins

Protease-activated receptor 2 (PAR2) is a G protein-coupled receptor which mediates a variety of functions in the skin including cutaneous inflammation. SLIGKV-NH(2) , an agonist peptide for PAR2, enhanced the interleukin (IL)-17-induced production of two CXC chemokines, CXCL1 (GRO-α) and CXCL8 (IL-8), in normal human epidermal keratinocytes (NHEK) in a concentration-dependent manner. The enhanced production of those chemokines was suppressed by a PAR2-specific siRNA. The SLIGKV-NH(2) -induced production of both CXCL1 and CXCL8 was markedly reduced by cyclosporine A. The enhanced production of CXCL1 was suppressed by 1α, 24R-dihydroxyvitamin D(3) , an active form of vitamin D(3) , and weakly by glucocorticoids, dexamethasone and clobetasol propionate, whereas production of CXCL8 was not altered by any of those receptor agonists. In psoriatic skin, the thickened upper spinous layer of the epidermis was positive for PAR2 protein and the expression of the IL17A mRNA was increased. These results suggest that the IL-17-induced pro-inflammatory reaction is enhanced by the activation of PAR2 in keratinocytes, and that the effect of PAR2 is differentially modulated by cyclosporine A, the active form of vitamin D(3) and glucocorticoids.

Topics: Cells, Cultured; Chemokine CXCL1; Clobetasol; Cyclosporine; Dexamethasone; Dihydroxycholecalciferols; Glucocorticoids; Humans; Inflammation Mediators; Interleukin-17; Interleukin-8; Keratinocytes; Oligopeptides; Psoriasis; Receptor, PAR-2; RNA, Messenger; RNA, Small Interfering

Topics: Adult; Aged; Corticotropin-Releasing Hormone; Dermatitis, Atopic; Disease Progression; Female; Gene Expression Regulation; Genetic Association Studies; Humans; Inflammation Mediators; Interleukin-8; Male; Mast Cells; Middle Aged; Psoriasis; Receptors, Corticotropin-Releasing Hormone; Skin; Vascular Endothelial Growth Factor A; Young Adult

Psoriasis is a benign, chronic skin disease characterized by keratinocyte hyperproliferation and abnormal differentiation. Curcumin, a selective phosphorylase kinase inhibitor, is a natural phytochemical present in turmeric. Curcumin has been confirmed to have anti-inflammatory properties as well as the ability to inhibit proliferation and decrease the expression of pro-inflammatory cytokines in psoriatic keratinocytes. However, the pro-apoptotic effect of curcumin in keratinocytes remains unclear. In the present study, we investigated the effect of curcumin on apoptosis induction in TNF-α-treated HaCaT cells. These results show that curcumin exhibited a significant pro-apoptotic effect on HaCaT cells only in the presence of TNF-α and/or TRAIL. The pro-apoptotic effect of curcumin resulted from the increased expression of TRAIL-R1/R2 and the decreased expression of anti-apoptotic proteins. Our results indicate that both curcumin and TNF-α up-regulated the expression of TRAIL-R1/R2. In addition, the expression of anti-apoptotic proteins (IAP1, IAP2, Bcl-X(L)) was up-regulated by TNF-α but suppressed by curcumin in HaCaT cells. Because these proteins are regulated by NF-κB, we examined the role of curcumin in NF-κB activation. As expected, curcumin inhibited TNF-α-induced activation of NF-κB, including NF-κB-P65. Curcumin also inhibited the TNF-α-induced production of IL-6/IL-8 in HaCaT cells. These results imply that curcumin-induced apoptosis of HaCaT cells only occurs when TNF-α or/and TRAIL are present. Therefore, we believe that curcumin is able to reverse the anti-apoptotic function of TNF-α in HaCaT cells and thus expect curcumin to be successful in the treatment of psoriasis.

Topics: Apoptosis; Cell Line; Curcumin; Humans; Interleukin-6; Interleukin-8; Keratinocytes; NF-kappa B; Psoriasis; Receptors, TNF-Related Apoptosis-Inducing Ligand; TNF-Related Apoptosis-Inducing Ligand; Tumor Necrosis Factor-alpha; Up-Regulation

IL-33, a member of the IL-1 family, is implicated in type 2 T helper cell immune reactions and acts as an "alarmin" to induce activation of dendritic cells in response to external stimuli. We investigated the effect of inflammatory cytokines on IL-33 expression in normal human epidermal keratinocytes. IFN-γ dose- and time-dependently induced IL-33 expression in protein and mRNA; this was dependent on extracellular signal-regulated kinase, p38, EGFR, and JAK phosphorylation. Combined IFN-γ and tumor necrosis factor (TNF)-α treatment induced expression of a 20-kDa band corresponding to mature IL-33, which was abolished by the addition of a calpain inhibitor. The addition of the inhibitor to IFN-γ and TNF-α-stimulated cells also induced strong expression of a 25-kDa band. Small interference (si) RNA for IL-33 abolished expression of the smaller bands and the 30-kDa IL-33 band, suggesting that these IL-33 forms were IL-33 transcription products. Recombinant IL-33 added in the medium induced IL-8 production, and RNA knockdown by siRNA enhanced IL-8 expression, suggesting its dual role as a cytokine and a nuclear factor. These results indicate that IL-33 has a role in inflammatory skin diseases, in which IFN-γ and TNF-α are present in high levels.

Topics: Calpain; Cells, Cultured; Dermatitis, Atopic; Epidermal Cells; Epidermis; Foreskin; Gene Expression; Humans; Infant, Newborn; Interferon-gamma; Interleukin-33; Interleukin-8; Interleukins; Keratinocytes; Lichen Planus; Male; MAP Kinase Signaling System; Psoriasis; RNA Interference; Skin Diseases; Tumor Necrosis Factor-alpha

Psoriasis, characterized by circumscribed, red, thickened plaques with an overlying silver-white scale, is a common T-cell-mediated chronic inflammatory skin disease. Although hydrogen sulfide (H(2)S) has been shown to be a signaling molecule with both pro- or anti-inflammatory effects, its relationship with psoriasis has not been elucidated. In the present study, 15 patients with chronic progressive psoriasis and 15 healthy volunteers were investigated. Serum H(2)S levels in psoriasis patients were significantly lower than those of healthy controls (16.69 ± 5.47 μM vs. 34.5 ± 6.39 μM). In contrast, serum levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and interleukin-8 (IL-8) were significantly higher in psoriasis patients than healthy controls (22.88 ± 6.24 pg/ml vs. 12.07 ± 3.68 pg/ml; 61.47 ± 8.21 pg/ml vs. 31.54 ± 13.73 pg/ml; and 39.43 ± 8.56 pg/ml vs. 20.55 ± 6.45 pg/ml, respectively). The serum H(2)S levels negatively correlated with clinical disease severity. Furthermore, treatment of HaCaT human keratinocytes with TNF-α increased the levels of nitric oxide (NO), IL-6 and IL-8 (32.21 ± 5.71 μM vs. 3.22 ± 0.98 μM; 203.96 ± 13.16 pg/ml vs. 13.57 ± 3.75 pg/ml; and 301.24 ± 30.17 pg/ml vs. 29.06 ± 10.91 pg/ml, respectively) in the culture media. Exogenous H(2)S inhibited the TNF-α-mediated upregulation of NO, IL-6 and IL-8 in a dose-dependent manner. In addition, H(2)S inhibited TNF-α-mediated activation of p38, extracellular-signal-regulated kinase and nuclear factor kappa B. In conclusion, H(2)S may play a protective role in the pathogenesis of psoriasis. H(2)S-releasing agents may be promising therapeutics for psoriasis.

Topics: Anti-Inflammatory Agents; Blotting, Western; Cell Line; DNA Primers; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Humans; Hydrogen Sulfide; Interleukin-6; Interleukin-8; Keratinocytes; Nitric Oxide; Nitrites; Psoriasis; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Tetrazolium Salts; Thiazoles; Tumor Necrosis Factor-alpha

A number of studies argue in favour of an important role of microbial colonization, in particular of Staphylococcus aureus, in triggering atopic dermatitis (AD) flare-up and psoriasis, in particular through the superantigenic properties of toxins generated by S. aureus.. The aim of this study was to assess the efficacy of a 3-week Avène hydrotherapy on the skin surface of patients suffering from psoriasis or atopic dermatitis.. Skin samples were taken from healthy subjects or atopic (n = 18) or psoriatic patients (n = 39) undergoing hydrotherapy at Avène at the beginning (D0) and the end of treatment (D18). The severity of the dermatosis was evaluated according to SCORing Atopic Dermatitis (SCORAD) or Psoriasis Area Severity Index (PASI) scores at D0 and D18. Marker of inflammation interleukin 8 (IL-8), S. aureus colonization (protein A) and enterotoxins were assessed in skin samples using RT-PCR.. At D0, significant differences were observed between healthy subjects and atopic or psoriatic patients in all the parameters evaluated (IL-8, protein A). At the end of the hydrotherapy, a significant decrease in SCORAD was associated with a significant reduction of IL-8, S. aureus colonization and enterotoxin D in patients with atopic dermatitis. Similarly, a significant decrease in PASI was associated with a significant reduction of IL-8, S. aureus colonization and enterotoxin N in patients with psoriasis.. This study demonstrates the positive effects of Avène hydrotherapy on the skin of patients suffering from chronic dermatosis, with decreased inflammation and reduced colonization by S. aureus.

Topics: Adult; Child; Child, Preschool; Dermatitis, Atopic; Enterotoxins; Humans; Hydrotherapy; Infant; Interleukin-8; Mineral Waters; Psoriasis; Severity of Illness Index; Staphylococcal Protein A; Staphylococcal Skin Infections; Staphylococcus aureus; Statistics, Nonparametric; Treatment Outcome; Young Adult

High-mobility group box-1 (HMGB1) plays important roles in inflammation, immune responses, and tumor progression. Since HMGB1 and its components have been shown to be mediators of a number of diseases but several sources of recombinant HMGB1 showed controversial biological activity, it is important to obtain recombinant HMGB1 with properties that resemble the native protein. For this purpose, we cloned genes coding for human HMGB1 and its active components A box and B box by PCR and inserted the cloned genes into pET28a vectors for transformation of Escherichia coli BL21. The E. coli expressed proteins were then purified with a Ni(2+)-NTA column and the endotoxin content was removed. Recombinant human HMGB1 (rhHMGB1) and its B box thus obtained stimulated, but A box inhibited, the production of the chemokine CXCL8/IL-8 by THP-1 monocytic cell line. We also used purified rhHMGB1 to immunize rabbits and generated potent anti-sera, which was capable of neutralizing the activity of rhHMGB1 in vitro and detecting the increased HMGB1 expression in inflammatory tissues in mice and humans. Thus, we have established essential means to produce biologically active rhHMGB1 that will facilitate us to study its role in diseases and to explore its potential as a therapeutic agent.

Topics: Animals; Cell Line; Cloning, Molecular; Concanavalin A; Escherichia coli; HMGB1 Protein; Humans; Immune Sera; Immunization; Inflammation; Interleukin-8; Liver Failure, Acute; Mice; Monocytes; Psoriasis; Rabbits; Recombinant Proteins; Transformation, Bacterial

Topics: Anti-Inflammatory Agents, Non-Steroidal; Cell Line; Cytokines; Epidermis; Fibroblasts; Gene Expression Regulation; Humans; Inflammation; Interleukin-6; Interleukin-8; Keratinocytes; Lipoxins; Models, Biological; Psoriasis; Skin

Psoriatic plaques have been shown to contain increased levels of pro-inflammatory cytokines. Also, serum levels of several cytokines have been reported elevated in psoriatic patients. It is postulated that changes in cytokine production both locally and systemically could be useful in monitoring disease activity. The aim of this study was to evaluate serum cytokine profile of interleukin (IL)-8, γ-interferon (IFN-γ) and tumor necrosis factor-α (TNF-α) in Egyptian psoriatic patients by enzyme-linked immunosorbent assay (ELISA) technique and to correlate these levels with disease severity. We analyzed serum samples from 60 Egyptian patients (31 females and 29 males) with a mean age of 40.2 ± 17.4 years with active psoriasis, and 21 healthy volunteers for major T-helper type 1 cytokines using the ELISA technique. The disease severity, including erythema, induration and scales, was assessed by Psoriasis Area and Severity Index (PASI) score. TNF-α and IFN-γ were markedly elevated in all sera from psoriatic patients. TNF-α was found a more efficient predictor for disease severity than IL-8 and IFN-γ using three receiver-operator curves with accuracy. IL-8 was also moderately elevated and correlated with the age of patients (r = 0.28). We have obtained evidence that TNF-α in our study was found to be more useful than the other two tested cytokines, IL-8 and IFN-γ as a follow-up marker for monitoring disease severity in Egyptian psoriatic patients. A positive correlation between lL-8 and the age of the patients was also noted.

Topics: Adolescent; Adult; Aged; Biomarkers; Child; Egypt; Female; Humans; Interferon-gamma; Interleukin-8; Male; Middle Aged; Psoriasis; Severity of Illness Index; Tumor Necrosis Factor-alpha; Young Adult

Semaphorin 7A (Sema7A) expressed on activated T cells stimulates cytokine production in monocytes through its receptor, α1β1 integrin.. To study the significance of Sema7A expressed on keratinocytes in skin inflammation where interaction between keratinocytes and β1-integrin expressing inflammatory cells, such as monocytes, takes place.. The regulation of Sema7A expression on keratinocytes by various cytokines was studied by flow cytometry and immunoblot. β1-integrin expressing human monocyte cell line, THP-1 cells, were co-cultured with paraformaldehyde-fixed normal human epidermal keratinocytes (NHK) and IL-8 production by THP-1 cells was studied. The significance of β1-integrin or Sema7A within this cell interaction was examined by the experiments using β1-integrin blocking antibody or Sema7A siRNA.. IFN-γ and TNF-α slightly increased Sema7A expression, while IL-4 decreased it. Among cytokines tested, TGF-β1 most strikingly increased the Sema7A expression on NHK. When NHK was stimulated by TGF-β1, paraformaldehyde-fixed, and co-cultured with THP-1 cells, IL-8 production by THP-1 cells was increased compared to THP-1 cells only. When THP-1 cells were pretreated with β1-integrin blocking antibody, this increase in IL-8 production by THP-1 cells was inhibited. Likewise, when NHK were pretreated with Sema7A siRNA before fixation and co-cultured with THP-1 cells, increase in IL-8 production by THP-1 cells was inhibited.. Our results suggest that Sema7A on keratinocytes and β1-integrin on monocytes contribute to monocyte activation by keratinocytes within skin inflammation, such as psoriasis or wound.

Topics: Antigens, CD; Coculture Techniques; Cytokines; DNA Primers; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; GPI-Linked Proteins; Humans; Inflammation; Integrin beta1; Interleukin-8; Keratinocytes; Monocytes; Psoriasis; RNA, Small Interfering; Semaphorins; Wound Healing

Sulfur is able to penetrate the skin, and a sulfur-rich balneotherapy has been suggested to be effective in the treatment of psoriasis. Psoriasis is now considered a genetically programmed, immune-mediated, inflammatory disease, in which intralesional T lymphocytes trigger keratinocytes to proliferate and perpetuate the disease process. Interleukin (IL)-17 and IL-22 produced by Th1/Th17 lymphocytes induce IL-8 secretion by keratinocytes, a key event in the pathogenesis of the disease. It is now clear that mitogen-activated protein kinase (MAPK) (extracellular signal-regulated kinases (ERK) 1 and 2) activity is required for IL-17-induced IL-8 synthesis by keratinocytes, and, in fact, MAPK activity is increased in lesional psoriatic skin. Here, we demonstrate both in vitro and in vivo on primary psoriatic lesions that pharmacological inhibitors of ERKs as well as hydrogen sulfide not only reduce the basal expression and secretion of IL-8, but also interfere with IL-17- and IL-22-induced IL-8 production. These observations, together with the known anti-inflammatory activity of H₂S, are relevant to understanding some previously unexplained biological effects exerted by sulfur therapy.

Topics: Aged; Cell Line; Extracellular Signal-Regulated MAP Kinases; Humans; Hydrogen Sulfide; Interleukin-17; Interleukin-22; Interleukin-8; Interleukins; Keratinocytes; Male; MAP Kinase Signaling System; Phosphorylation; Psoriasis

Plasmacytoid dendritic cells (pDC) are present in inflammatory skin lesions, in particular, in psoriasis. In such lesions, the inflammatory mediator histamine is also detected in high amounts. We therefore investigated a possible interaction of pDC with histamine, especially via the most recently described histamine H(4) receptor (H(4)R). We detected the expression of the H(4)R on pDC in the blood and in lesional psoriasis skin. Interestingly, compared with healthy controls and patients with atopic dermatitis, pDC from the blood of psoriasis patients expressed the highest levels of the H(4)R, which was even more upregulated on stimulation with IFN-γ and CpG. After activation of the H(2)R and H(4)R on pDC, we observed downregulation of CpG-induced production of tumor necrosis factor α, IFN-α, and CXCL8, but not of the chemokine CXCL10. Histamine-induced downregulation of cytokine production was more pronounced in pDC derived from psoriasis patients. Furthermore, we observed F-actin polymerization and active migration of pDC in response to H(4)R agonist stimulation. Taken together, our results indicate that the H(4)R is highly expressed on pDC in psoriasis and influences cytokine production and migration of pDC. Therefore, the H(4)R alone or in combination with the H(2)R might be a promising therapeutic target in psoriasis.

Topics: Blood Buffy Coat; Cell Movement; Cells, Cultured; Dendritic Cells; Dermatitis, Atopic; Gene Expression; Histamine; Humans; Interferon-alpha; Interferon-gamma; Interleukin-13; Interleukin-8; Oligonucleotides; Psoriasis; Receptors, G-Protein-Coupled; Receptors, Histamine; Receptors, Histamine H4; Skin; Tumor Necrosis Factor-alpha

Psoriasis patients are frequently associated with metabolic syndromes. Such associations are possibly mediated by adipokines. We investigated the in vitro effects of visfatin (an adipokine) on chemokine expression in human keratinocytes. Normal human keratinocytes were incubated with visfatin, and their chemokine production was analyzed by ELISA and RT-PCR. Visfatin enhanced TNF-α-induced CXC chemokine ligand (CXCL) 8, CXCL10, and CC chemokine ligand (CCL) 20 secretion and mRNA expression in keratinocytes, although visfatin alone was ineffective. A small interfering RNA against nuclear factor-κB (NF-κB) p65 suppressed the visfatin-induced production of CXCL8, CXCL10, and CCL20 whereas a small interfering RNA against signal transducer and activator of transcription (STAT) 3 suppressed CXCL8 induction. This indicates the involvement of NF-κB in CXCL8, CXCL10, and CCL20 induction by visfatin and the involvement of STAT3 in CXCL8 induction. Visfatin alone increased the transcriptional activity and tyrosine phosphorylation of STAT3, which was suppressed by Janus kinase (JAK) 2 inhibitor. Visfatin enhanced basal and TNF-α-induced NF-κB activity and inhibitory κB (IκB) α phosphorylation, which was suppressed by IκB kinase inhibitor. Visfatin induced the tyrosine and serine phosphorylation of JAK2 and IκB kinase α/β, respectively. Intraperitoneal injection of visfatin elevated mRNA and protein levels of CXCL1, CXCL10, and CCL20 in murine skin. These results suggest that visfatin enhances CXCL8, CXCL10, and CCL20 production in human keratinocytes and homologous chemokine production in murine skin. Visfatin may induce the infiltration of type 1 or type 17 helper T cells or neutrophils to the skin via chemokine induction and thus link metabolic syndromes to psoriasis.

Topics: Animals; Cells, Cultured; Chemokine CCL20; Chemokine CXCL10; Female; Humans; I-kappa B Kinase; Interleukin-8; Janus Kinase 2; Keratinocytes; Mice; Mice, Inbred BALB C; NF-kappa B; Niacinamide; Nicotinamide Phosphoribosyltransferase; Pentosyltransferases; Psoriasis; Receptor, Insulin; STAT3 Transcription Factor; Tumor Necrosis Factor-alpha

Tumour necrosis factor-like weak inducer of apoptosis (TWEAK), a member of the tumour necrosis factor (TNF) family, has been implicated as a proinflammatory cytokine in many types of autoimmune and infectious diseases. However, information about TWEAK in dermatological diseases is limited. Herein, we investigated the role of TWEAK in patients with Henoch-Schonlein purpura (HSP) and the ability of TWEAK on chemokine production in the human dermal microvascular endothelial cell line (HMEC-1). Serum TWEAK levels in patients with HSP, together with patients with psoriasis vulgaris (PV) and atopic dermatitis (AD), were detected by enzyme-linked immunosorbent assay (ELISA). HMEC-1 cells were treated with TWEAK at concentrations ranging from 1 ng/ml to 100 ng/ml. Serum levels of TWEAK were elevated in patients with HSP in the acute stage but not in patients with PV or AD. Moreover, TWEAK levels were correlated with the severity of HSP. TWEAK markedly induced CCL5 and CXCL8 production at both mRNA and protein levels in HMEC-1 cells. In addition, TWEAK-stimulated HMEC-1 supernatant enhanced HL-60 or human acute monocytic leukaemia cell line (THP-1) cell migration. Finally, Western blot data revealed that TWEAK can induce rapid phosphorylation of inhibitor of κB-α (IκBα) in HMEC-1 cells. In conclusion, we show that serum levels of TWEAK were elevated in patients with acute stage HSP. TWEAK may act as a regulator of nuclear factor-κB (NF-κB) activation and chemokine production in human dermal microvascular endothelial cells, thus promoting leucocyte migration in cutaneous vasculitis.

Topics: Adolescent; Adult; Apoptosis; Blotting, Western; Case-Control Studies; Cells, Cultured; Chemokine CCL5; Child; Child, Preschool; Cytokine TWEAK; Dermatitis, Atopic; Endothelial Cells; Female; Gene Expression; Humans; I-kappa B Proteins; IgA Vasculitis; Inflammation; Interleukin-8; Male; NF-kappa B; NF-KappaB Inhibitor alpha; Phosphorylation; Polymerase Chain Reaction; Psoriasis; RNA, Messenger; Signal Transduction; Skin; Tumor Necrosis Factors

Mast cells are increasingly present in the lesional skin of chronic skin inflammatory diseases including psoriasis and basal cell carcinoma (BCC). It has previously been shown that proteinase-activated receptor (PAR)-2 is expressed by mast cells, and tryptase is a potent activator of this receptor. In this study, skin biopsies from both healthy-looking and lesional skin of patients with psoriasis and superficial spreading BCC were collected and the expression of PAR-2 immunoreactivity in tryptase-positive mast cells was analysed. PAR-2 expression was confirmed in vitro in different mast cell populations. Cord-blood derived mast cells (CBMC) were stimulated with a PAR-2 activating peptide, 2-furoyl-LIGRLO-NH(2). Consequently, IL-8 and histamine production was analysed in the supernatants. We observed a significant increase in the percentage of mast cells expressing PAR-2 in the lesional skin of psoriasis and BCC patients compared with the healthy-looking skin. HMC-1.2, LAD-2 and CBMC mast cells all expressed PAR-2 both intracellularly and on the cell surface. CBMC activation with the PAR-2 activating peptide resulted in an increased secretion of IL-8, but no histamine release was observed. Furthermore, both PAR-2 and IL-8 were co-localized to the same tryptase-positive mast cells in the lesional BCC skin. These results show that mast cells express increased levels of PAR-2 in chronic skin inflammation. Also, mast cells can be activated by a PAR-2 agonist to secrete IL-8, a chemokine which can contribute to the progress of inflammation.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Basal Cell; Female; Humans; Interleukin-8; Male; Mast Cells; Middle Aged; Psoriasis; Receptor, PAR-2; Skin Neoplasms; Up-Regulation; Young Adult

The strong but complex genetic background suggests that inherent and intrinsic rather than exogenous factors have a key role in immunopathogenesis of psoriasis. It is reasonable to speculate that the dysfunctional activity of psoriatic T cells may partly originate from the abnormal haematopoietic cells.. To test if T cells originated from haematopoietic progenitor cells in psoriasis patients display functional alternations similar to previously reported abnormalities of circulating T cells.. Bone marrow CD34(+) haematopoietic cells were isolated from psoriatic patients with family history and healthy subjects, and differentiated into T cells in vitro in the thymic stromal co-culture system. These cells were further subjected to functional comparisons such as in vitro proliferation, secretion of cytokines such as IL-4, IL-8 and IFN-gamma, and inducing the production of C-myc, Bcl-xL, and Ki67 proteins in human keratinocytes.. While bone marrow-derived CD34(+) cells from both patients and healthy volunteers developed into mature T cells within weeks in the thymic environment in vitro, the differentiated T cells from psoriatic patients showed higher proliferation and stronger capacity to secret TH1 cytokines in response to streptococcal superantigen. The differentiated T cells from psoriatic patients, but not from normal controls, induced overexpression of C-myc and Ki67, but not Bcl-XL, in keratinocytes.. T cells differentiated from CD34(+) cells of psoriatic patients, but not normal controls, are functionally similar to psoriatic circulating T cells, suggesting that the dysfunctional activity of T cells in psoriatic patients can be traced back to the early development of haematopoietic cells.

Topics: Adult; Antigens, CD34; bcl-X Protein; Bone Marrow; Case-Control Studies; Cell Differentiation; Cell Proliferation; Coculture Techniques; Family Health; Female; Hematopoietic Stem Cells; Humans; Interferon-gamma; Interleukin-8; Keratinocytes; Male; Middle Aged; Proto-Oncogene Proteins c-myc; Psoriasis; Stromal Cells; T-Lymphocytes

Psoriasis is an immune-mediated disease typically associated with cutaneous neutrophilic infiltration and Munro microabscesses. Interleukin (IL)-8 is one of the main neutrophil-attracting chemokines. Although keratinocytes have traditionally been considered to be the principal source of IL-8 in psoriasis, we present data that suggest that cutaneous lymphocyte associated antigen (CLA) + T lymphocytes synthesize this cytokine.. Six patients with psoriasis and 6 healthy controls were studied. Immunomagnetic separation was used to isolate CLA+ and CLA- T lymphocytes and IL-8 and interferon (IFN)-gamma production was quantified for each cell subpopulation using enzyme-linked immunosorbent assay. Finally, gene expression of IL-8 was analyzed by reverse transcriptase-polymerase chain reaction.. CLA+ and CLA- T lymphocytes from patients with psoriasis and from controls showed a significantly increased production of IFN-gamma when activated, whereas only activated CLA+ T lymphocytes (from patients and controls) synthesized IL-8. The higher level of expression of IL-8 and IFN-gamma by CLA+T lymphocytes in comparison to CLA- cells was confirmed.. Previous studies have confirmed IL-8 production by T lymphocytes in inflammatory skin diseases with neutrophil-rich infiltrates, such as acute generalized exanthematous pustulosis, Behçet disease, and pustular psoriasis. We have confirmed the role of the subset of T lymphocytes with skin tropism (CLA+) in IL-8 production in nonpustular psoriasis.

Topics: Antigens, Differentiation, T-Lymphocyte; Antigens, Neoplasm; Chemotaxis; Computer Systems; Gene Expression Regulation; Humans; Immunologic Memory; Interferon-gamma; Interleukin-8; Lymphocyte Activation; Membrane Glycoproteins; Neutrophils; Psoriasis; Reverse Transcriptase Polymerase Chain Reaction; Skin; T-Lymphocyte Subsets

Several cross-sectional studies have shown that different cytokines and growth factors are enhanced in psoriasis.. We aimed to understand the role/relation of interleukin (IL)-22, IL-17, IL-23, IL-8, vascular endothelial growth factor (VEGF) and tumour necrosis factor (TNF)-α in psoriasis vulgaris, addressing their levels and changes before, during and after psoralen-ultraviolet A (PUVA) and narrowband ultraviolet B (NB-UVB) treatment.. A cross-sectional and a longitudinal study (n = 34) - before (T0) and at 3 (T3), 6 (T6) and 12 (T12) weeks of NB-UVB and PUVA therapy - were performed; 17 patients started NB-UVB and 17 PUVA, and IL-22, IL-17, IL-23, IL-8, TNF-α and VEGF levels were evaluated.. At T0, compared with controls (n = 20), all the parameters were significantly higher in patients, except for TNF-α. Both NB-UVB and PUVA treatment gave, at T3, a significant decrease in TNF-α and IL-23; IL-22 and IL-17 decreased significantly at T6; all parameters and Psoriasis Area and Severity Index decreased significantly at T12. However, in both groups, at T12, VEGF was still significantly higher than control.. Psoriasis seems to be a complex disease in which the cytokine network is disturbed, namely in levels of IL-22, IL-17, IL-23, IL-8, TNF-α and VEGF. NB-UVB and PUVA follow-up studies suggested that the reduction in the IL-23/Th17 axis might be important in the pathogenic mechanisms of psoriasis. Further follow-up studies of patients with psoriasis treated with these and other therapies could be very helpful for the understanding of the disturbance in the cytokine network in psoriasis and indirectly in its pathogenesis.

Topics: Adult; Cross-Sectional Studies; Cytokines; Female; Humans; Interleukin-17; Interleukin-22; Interleukin-23; Interleukin-8; Interleukins; Longitudinal Studies; Male; Middle Aged; Psoriasis; PUVA Therapy; Severity of Illness Index; Tumor Necrosis Factor-alpha; Ultraviolet Therapy; Vascular Endothelial Growth Factor A

Psoriasis is a common skin disease affecting about 1-3% of the world population. Many types of cells, including lymphocytes, dendritics APCs (antigen presenting cells), NKT (natural killer T) cells, neutrophils, keratinocytes and fibroblasts are involved in the pathogenesis of psoriasis.The aim of our study was to assess in psoriatic patients the production of IL-8, IL-10 and IL-12 cytokines by neutrophils, fibroblasts and fibroblasts - neutrophils interaction.. The production of IL-8, IL-10 and IL-12 cytokines was evaluated in supernatants after cells incubation for 21 h in culture medium alone and in medium in the presence of IL-8 or TNF-α. Concentrations of IL-8, IL-10, IL-12 were measured by enzyme-linked immunosorbent assay (ELISA) method using commercially available kits.. Our results demonstrate that fibroblasts are not able to produce IL-10 and IL-12 but they generate IL-8. The amount of IL-8 depends on the site of derivation of fibroblasts and on the stimuli. Neutrophils released IL-8, IL-12 (at a lower level in psoriatic patients) and IL-10 but only in the case of healthy donors and at a very low concentration. Moreover, we observed higher concentrations of IL-12 and IL-8 in supernatants as a result of fibroblasts-neutrophils interaction in psoriatic patients.. Our results suggest that fibroblasts take part in the inflammatory reaction in psoriasis via cytokines or direct interaction with neutrophils. Fibroblasts probably do not exert any anti-inflammatory effect.

Topics: Adult; Female; Fibroblasts; Humans; Interleukin-10; Interleukin-12; Interleukin-8; Male; Middle Aged; Neutrophils; Psoriasis

IL-17F is known to be involved in many inflammatory diseases, but its role in skin diseases has not been fully examined. Because IL-8 is involved in many skin diseases such as psoriasis, we investigated the production of IL-8 in normal human epidermal keratinocytes (NHEKs) stimulated by IL-17F, tumor necrosis factor-alpha (TNF-alpha), IL-17A, and control using real-time PCR and ELISA. The results showed that IL-17F induced production of IL-8 in NHEKs in a time-dependent manner. Interestingly, the amounts of IL-8 stimulated by IL-17F were much higher than those stimulated by TNF-alpha or IL-17A. Next, we confirmed that selective mitogen-activated protein kinase kinase inhibitors significantly inhibited IL-17F-induced IL-8 production. Moreover, mouse skin intradermally injected with IL-17F expressed high level of IL-8 mRNA and induced ERK1/2 phosphorylation. Histological examination of mouse skin that was injected with IL-17F revealed marked neutrophilia in dermis and the infiltration was significantly inhibited by anti-IL-8 antibody. Finally, IL-17F expression in skin biopsy samples from psoriasis patients were examined by western blotting and ELISA. IL-17F was upregulated in lesional psoriatic skin compared with nonlesional skin. These results indicate that IL-17F may be involved in psoriasis via, in part, the activation of ERK1/2 and the induction of IL-8 in keratinocytes.

Topics: Animals; Epidermis; Female; Gene Expression Regulation; Humans; Interleukin-17; Interleukin-8; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Neutrophils; Phosphorylation; Psoriasis; RNA, Messenger

Osteopontin (OPN) is a phosphorylated acidic glycoprotein produced by cells of the immune system, epithelial tissue, smooth muscle cells, osteoblasts, and tumor cells. OPN interacts with integrins and CD44 to enhance Th1 and inhibit Th2 cytokine expression. The involvement of this molecule in the onset of psoriasis has not previously been studied. Here, we demonstrate that OPN is expressed in peripheral blood mononuclear cells and in skin biopsies of psoriatic patients. The study was conducted on 30 patients affected with plaque psoriasis, and on 11 healthy donors. Two blood samples and two skin samples from patients affected with atopic dermatitis were used as control for Th2 typical inflammatory skin disease. The analysis of IL-1beta, IFN-gamma, TauNuF-alpha, IL-8, and ICAM-1 showed the characteristic Th1 pattern in all the psoriatic blood and skin samples analyzed. This study offers an opportunity for understanding inflammation in psoriasis and supports the hypothesis that OPN could represent a potential target for therapeutic intervention in psoriatic patients.

Topics: Biopsy; Gene Expression Regulation; Humans; Hyaluronan Receptors; Inflammation; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-1beta; Interleukin-8; Leukocytes, Mononuclear; Osteopontin; Psoriasis; Skin; Th1 Cells; Th2 Cells

Antimicrobial peptides (AMPs) are strongly expressed in lesional skin in psoriasis and play an important role as proinflammatory "alarmins" in this chronic skin disease. Vitamin D analogs like calcipotriol have antipsoriatic effects and might mediate this effect by changing AMP expression. In this study, keratinocytes in lesional psoriatic plaques showed decreased expression of the AMPs beta-defensin (HBD) 2 and HBD3 after topical treatment with calcipotriol. At the same time, calcipotriol normalized the proinflammatory cytokine milieu and decreased interleukin (IL)-17A, IL-17F and IL-8 transcript abundance in lesional psoriatic skin. In contrast, cathelicidin antimicrobial peptide expression was increased by calcipotriol while psoriasin expression remained unchanged. In cultured human epidermal keratinocytes the effect of different vitamin D analogs on the expression of AMPs was further analyzed. All vitamin D analogs tested blocked IL-17A induced HBD2 expression by increasing IkappaB-alpha protein and inhibition of NF-kappaB signaling. At the same time vitamin D analogs induced cathelicidin through activation of the vitamin D receptor and MEK/ERK signaling. These studies suggest that vitamin D analogs differentially alter AMP expression in lesional psoriatic skin and cultured keratinocytes. Balancing AMP "alarmin" expression might be a novel goal in treatment of chronic inflammatory skin diseases.

Topics: Anti-Infective Agents; Antimicrobial Cationic Peptides; Blotting, Western; Cathelicidins; Cells, Cultured; Gene Expression Regulation; Genes, Reporter; Humans; Interleukin-17; Interleukin-8; MAP Kinase Signaling System; Peptides; Polymerase Chain Reaction; Psoriasis; Receptors, Calcitriol; RNA Interference; Vitamin D

IL-20 cytokine subfamily members, including IL-19, IL-20, and IL-24, are highly expressed in psoriatic skin lesions. Here, we demonstrate that psoriasis mediators IL-17 and IL-22 synergistically induce the production of IL-20 subfamily proteins in cultured human keratinocytes. Interestingly, expression of the IL-22 receptor (IL-22R) also increased in epidermal lesions versus normal skin. IL-22R over-expression using an adenoviral vector to mimic psoriatic conditions in cultured keratinocytes significantly enhanced IL-17- and IL-22-induced production of IL-20 subfamily cytokines. Furthermore, IL-17 and IL-22 coordinately enhanced MIP-3alpha, IL-8, and heparin-binding EGF-like growth factor (HB-EGF) production, depending on the amount of IL-22R expression. Additionally, because IL-20 and IL-24 share the IL-22R with IL-22, the function of IL-20 and IL-24 was also increased. IL-20 and IL-24 have effects similar to that of IL-22; IL-24 showed more potent expression than IL-20. A combination of IL-24 and IL-17 increased the production of MIP-3alpha, IL-8, and HB-EGF, as did a combination of IL-22 and IL-17. These data indicate that increased IL-22R expression in epidermal keratinocytes contributes to the pathogenesis of psoriasis through enhancing the coordinated effects of IL-22 and IL-17, inducing the production of the IL-20 subfamily, chemokines, and growth factors.

Topics: beta-Defensins; Cells, Cultured; Chemokine CCL20; Epidermis; Gene Expression; Heparin-binding EGF-like Growth Factor; Humans; Intercellular Signaling Peptides and Proteins; Interferon-gamma; Interleukin-10 Receptor beta Subunit; Interleukin-17; Interleukin-1alpha; Interleukin-22; Interleukin-8; Interleukins; Keratinocytes; Models, Biological; Phosphorylation; Psoriasis; Receptors, Interleukin; STAT3 Transcription Factor; Transduction, Genetic; Transforming Growth Factor alpha; Tumor Necrosis Factor-alpha

Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1) functions in both DNA repair and redox signaling, making it an attractive emerging therapeutic target. However, the role of APE1 in cutaneous inflammatory responses is largely unknown. In this study, we report that APE1 is a key upstream regulator in TLR2-dependent keratinocyte inflammatory responses. We found that nuclear expression of APE1 in epidermal layers was markedly up-regulated in psoriatic skin. APE1 was essential for the transcriptional activation and nuclear translocation of hypoxia-inducible factor-1alpha and NF-kappaB, both of which are crucial for inflammatory signaling in keratinocytes. Moreover, APE1 played a crucial role in the expression of TLR2-mediated inflammatory mediators, including TNF-alpha, CXCL8, and LL-37, in HaCaT cells and human primary keratinocytes. Silencing of APE1 attenuated cyclin D1/cyclin-dependent kinase 4 expression and phosphorylation of ERK1/2 and Akt, thereby affecting keratinocyte proliferation. Importantly, TLR2-induced generation of reactive oxygen species contributed to the nuclear translocation and expression of APE1, suggesting an autoregulatory circuit in which the subcellular localization of APE1 is associated with the production of APE1 per se through reactive oxygen species-dependent signaling. Taken together, these findings establish a role for APE1 as a master regulator of TLR2-dependent inflammatory responses in human keratinocytes.

Topics: Antimicrobial Cationic Peptides; Apoptosis; Cathelicidins; Cell Line; Cyclin D1; Cyclin-Dependent Kinase 4; DNA-(Apurinic or Apyrimidinic Site) Lyase; Epidermis; Extracellular Signal-Regulated MAP Kinases; Humans; Hypoxia-Inducible Factor 1; Inflammation; Interferon Inducers; Interleukin-8; Keratinocytes; NF-kappa B; Poly I-C; Proto-Oncogene Proteins c-akt; Psoriasis; Reactive Oxygen Species; RNA, Small Interfering; Signal Transduction; Toll-Like Receptor 2; Transcriptional Activation; Transfection; Tumor Necrosis Factor-alpha; Up-Regulation; Zymosan

IL-8 is a chemokine that has been implicated in a number of inflammatory diseases involving neutrophil activation. HuMab 10F8 is a novel fully human mAb against IL-8, which binds a discontinuous epitope on IL-8 overlapping the receptor binding site, and which effectively neutralizes IL-8-dependent human neutrophil activation and migration. We investigated whether interference in the cytokine network by HuMab 10F8 might benefit patients suffering from palmoplantar pustulosis, a chronic inflammatory skin disease. Treatment of patients with HuMab 10F8 was well tolerated and significantly reduced clinical disease activity at all five endpoints, which included a >or=50% reduction in the formation of fresh pustules. IL-8 neutralization was monitored at the site of inflammation by assessing exudates of palmoplantar pustulosis lesions. HuMab 10F8 sequestered IL-8 in situ, as observed by rapid dose-dependent decreases of IL-8 concentrations immediately following Ab infusion. These data demonstrate a critical role for IL-8 in the pathophysiology of palmoplantar pustulosis. HuMab 10F8 is capable of interrupting IL-8 activity in vivo and represents a candidate for treatment of inflammatory diseases and other pathological conditions associated with IL-8 overproduction.

Topics: Amino Acid Sequence; Animals; Antibodies, Monoclonal; Cells, Cultured; Epitopes; Humans; Immune Tolerance; Immunotherapy; Inflammation; Interleukin-8; Mice; Mice, Transgenic; Models, Molecular; Molecular Sequence Data; Neutrophils; Protein Binding; Protein Structure, Tertiary; Psoriasis; Time Factors

Oral acitretin is currently indicated for the treatment of severe psoriasis in adults, but its use is limited by systemic side effects and teratogenicity. Topical administration of acitretin may lessen the risk of systemic toxicity while increasing local bioavailability in the skin. The effects of topical acitretin on reconstructed human epidermis (RHE) and Rhino mice were investigated and compared to those of currently marketed topical retinoids: tretinoin and tazarotene. In acitretin-treated RHE cultures, there was a reduction in keratohyalin granules and filaggrin expression in the stratum granulosum, a loss of keratin 10 expression in the stratum spinosum, and an increase in keratin 19 expression in all viable cell layers. All retinoids showed similar signs of activity in RHE cultures. Furthermore, the release of pro-inflammatory cytokines IL-1alpha and IL-8 in RHE cultures was less pronounced with acitretin compared to tretinoin- and tazarotene-containing formulations, suggesting that acitretin may be less irritating. In Rhino mice, acitretin induced a local, dose-dependent reduction in utricle diameter after seven daily dermal doses. A similar effect was observed in tretinoin- and tazarotene-treated mice. Our data suggest that topical application of acitretin may have a therapeutic benefit in the local management of keratinization disorders.

Topics: Acitretin; Administration, Topical; Animals; Disease Models, Animal; Epidermis; Female; Filaggrin Proteins; Humans; Interleukin-1alpha; Interleukin-8; Keratin-10; Keratin-19; Keratolytic Agents; Male; Mice; Mice, Hairless; Nicotinic Acids; Psoriasis; Tretinoin

In psoriasis CD4+CD25+ regulatory T cells are functionally deficient. The imbalance between regulatory and effector T-cell functions is important for inducing psoriasis. It is reasonable to speculate that the dysfunctional activity of CD4+CD25+ regulatory cells may originate partly from the abnormal haematopoietic cells determined mainly by genetic background.. To test the hypothesis that haematopoietic stem cells are responsible for dysfunctional CD4+CD25+ regulatory cells in psoriasis.. Bone marrow-derived CD34+ haematopoietic cells from patients with psoriasis (with a family history of psoriasis) and from normal controls were differentiated into T cells in vitro. CD4+CD25+ T cells were isolated by an immunomagnetic bead method, and proliferation activity and capacity for cytokine secretion were determined. Furthermore, the ability of CD4+CD25+ T cells to suppress the proliferative responses of allogeneous peripheral blood CD4+CD25- effector T cells was assessed in vitro.. The differentiated CD4+CD25+ T cells of psoriatic origin showed similar characteristics to those of normal volunteers, including proliferation activity and secretion profile of the cytokines interleukin (IL)-2, IL-4, IL-8, IL-10 and interferon (IFN)-gamma. However, proliferation and secretion levels of the cytokines IL-2 and IL-10 for CD4+CD25+ cells of psoriatic CD34+ cell origin were significantly lower than those of normal controls in response to streptococcal superantigen (Strep-A). In particular, CD4+CD25+ T cells differentiated from psoriatic CD34+ cells were functionally insufficient to restrain effector T-cell proliferation.. CD4+CD25+ T cells differentiated in vitro from haematopoietic cells of patients with psoriasis are impaired in regulatory function. The dysfunction of psoriatic CD4+CD25+ T cells may be due to inherent genetic programming passed down from bone marrow-derived haematopoietic cells.

Topics: Adult; Case-Control Studies; Cell Differentiation; Cell Proliferation; Cytokines; Family Health; Female; Hematopoietic Stem Cells; Humans; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-4; Interleukin-8; Male; Middle Aged; Psoriasis; T-Lymphocytes, Regulatory

Neutrophils are the first cells arriving at sites of acute inflammation. On their way from blood to the site of inflammation, neutrophils have to adhere to endothelial cells (EC), to transverse the basement membrane and subsequently to travel through the interstitial matrix. Recently, we have shown that human Thy-1 is an alternate EC receptor for the leukocyte integrin Mac-1 that contributes to leukocyte recruitment to sites of inflammation, providing a new pathway for adhesion and transmigration of neutrophils. Here, we studied the effect of Thy-1-mediated adhesion on neutrophil functions. Binding of neutrophils to recombinant human Thy-1 stimulated the release of MMP-9 from neutrophils, resulting in their enhanced migration through collagen-IV and matrigel. Further, we showed that neutrophil interaction with Thy-1 stimulated secretion of CXCL8 and thus could support the attraction of additional neutrophils to inflammatory sites. Blocking experiments confirmed the pivotal roles of Thy-1 on activated dermal EC or fibroblasts and its counter receptor CD18 on neutrophils for the regulation of MMP-9 and CXCL8 release from neutrophils. Our results support the general concept that the function of 'adhesion molecules' in particular of human Thy-1, may not only be to provide mechanical support but also regulate neutrophil functions.

Topics: Antibodies, Monoclonal; CD18 Antigens; Cell Adhesion; Cell Communication; Cell Migration Assays, Leukocyte; Cell Movement; Cells, Cultured; Chemotaxis, Leukocyte; Coculture Techniques; Culture Media, Conditioned; Endothelial Cells; Fibroblasts; Humans; Interleukin-8; Isoantibodies; Matrix Metalloproteinase 9; Neutrophils; Psoriasis; Recombinant Proteins; RNA, Small Interfering; Skin; Thy-1 Antigens

The critical role of IL-17 has recently been reported in a variety of conditions. Since IL-17 deeply participates in the pathogenesis of psoriasis and keratinocyte production of certain cytokines, the involvement of T helper cell 17 (Th17) in atopic dermatitis (AD) is an issue to be elucidated. To evaluate the participation of Th17 cells in AD, we successfully detected circulating lymphocytes intracellularly positive for IL-17 by flow cytometry, and the IL-17+ cell population was found exclusively in CD3+CD4+ T cells. The percentage of Th17 cells was increased in peripheral blood of AD patients and associated with severity of AD. There was a significant correlation between the percentages of IL-17+ and IFN-gamma+ cells, although percentage of Th17 cells was not closely related to Th1/Th2 balance. Immunohistochemically, IL-17+ cells infiltrated in the papillary dermis of atopic eczema more markedly in the acute than chronic lesions. Finally, IL-17 stimulated keratinocytes to produce GM-CSF, TNF-alpha, IL-8, CXCL10, and VEGF. A marked synergistic effect between IL-17 and IL-22 was observed on IL-8 production. The number of Th17 cells is increased in the peripheral blood and acute lesional skin of AD. Th17 cells may exaggerate atopic eczema.

Topics: Adolescent; Adult; Aged; Case-Control Studies; CD4-Positive T-Lymphocytes; Cell Movement; Child; Dermatitis, Atopic; Dermis; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-17; Interleukin-22; Interleukin-8; Interleukins; Male; Middle Aged; Psoriasis; T-Lymphocytes, Helper-Inducer; Th1 Cells; Th2 Cells; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

The multifunctional cytokine tumor necrosis factor-alpha (TNF-alpha) is known to play an important role in inflammatory and immunological responses in human skin. Although it has been documented that reactive oxygen species (ROS) are involved in TNF-alpha-induced signaling pathways associated with certain inflammatory diseases, their role in TNF-alpha signaling cascades has not been examined in primary human keratinocytes used as a model of inflammatory skin disease and psoriasis. Employing a series of in vitro and in cellulo approaches, we have demonstrated that in primary human keratinocytes (i) TNF-alpha rapidly induces ROS generation, IkappaB degradation, NF-kappaB p65 nuclear translocation, and ultimately production of inflammatory cytokines; (ii) TNF-alpha-induced cytokine production is mediated both by the mammalian target of rapamycin signaling pathway via NF-kappaB activation and by ROS; (iii) TNF-alpha-dependent NF-kappaB activation (that is, IkappaB degradation and NF-kappaB p65 nuclear translocation) is not mediated by ROS; and (iv) a cell-penetrating derivative of the antioxidant enzyme, catalase, as well as taurine and N-acetyl-cysteine attenuate the TNF-alpha-induced production of cytokines. These latter results suggest that catalase and perhaps other antioxidants should be considered as part of a more specific and effective therapy for the treatment of inflammatory skin diseases, including psoriasis.

Topics: Acetylcysteine; Antioxidants; Catalase; Cells, Cultured; Free Radical Scavengers; Humans; Hydrogen Peroxide; I-kappa B Proteins; Interleukin-6; Interleukin-8; Keratinocytes; Male; Protein Kinases; Psoriasis; Reactive Oxygen Species; Signal Transduction; Sirolimus; Skin Diseases; Taurine; TOR Serine-Threonine Kinases; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Thermal balneotherapy with Comano's spa water (CW; Trentino, Italy) is beneficial for psoriasis and other skin disorders but its operative mechanisms are largely unknown. Previously, we showed that CW interferes with the production and secretion of IL-6 and various VEGF-A isoforms and with CK-16 expression by cultured human psoriatic keratinocytes. In this study, confluent cultures of epidermal keratinocytes isolated from the lesional areas of 9 psoriatic patients were exposed for 11-13 days to DMEM, whose chemicals had been dissolved in either deionised water (DW-DMEM, controls) or CW (CW-DMEM, treated cells), in order to assess the expression and secretion of TNF-alpha and IL-8 by such cells. The results gained by means of immunocytochemistry, Western immunoblotting (WB), and ELISA assays showed that CW exposure significantly down-regulated the intracellular levels of TNF-alpha, a key inducer of IL-8, IL-6, and other chemokines. However, no assayable TNF-alpha secretion occurred in keratinocyte-conditioned DW- and CW-DMEM samples. Moreover, the intracellular levels and secretion rates of IL-8 were also markedly reduced in the protein extracts and conditioned media of CW-DMEM-incubated keratinocytes. Notably, the most effective inhibition of IL-8 secretion was elicited by a 25% CW fraction in the DMEM. Altogether, our findings indicate that by attenuating at lesional skin sites the deregulated production and secretion of a cascade of several cytokines and chemokines (e.g. TNF- alpha, IL-8, IL-6, and various VEGF-A isoforms), and by offsetting the keratinocytes' abnormal differentiation program entailing CK-16 expression, CW balneotherapy may beneficially influence the clinical manifestations of psoriasis.

Topics: Balneology; Cells, Cultured; Down-Regulation; Humans; Interleukin-8; Italy; Keratinocytes; Psoriasis; Tumor Necrosis Factor-alpha; Water

Psoriasis is a chronic inflammatory cutaneous disorder. It is marked by aberrant epidermal and dermal expression of cytokines.. Evaluate the expression of proinflammatory cytokines in a particular severe form of psoriasis the psoriatic erythroderma.. We focused on intra-lesional cytokine gene expression in cutaneous biopsies of lesional site and their correspondent non lesional skin. On the whole, four healthy volunteers and thirty six patients were included in this study. Among these, three had a psoriatic erythroderma. Assuming that local production of cytokines may be approached by mRNA cytokine quantification, the expression of alpha tumour necrosis factor (TNFalpha) and interleukin-8 (IL-8) was analyzed by reverse transcription and real time quantitative polymerase chain reaction.. Under expression of all selected molecules in psoriatic erythroderma lesions was contrasted with the data obtained in the other psoriatic lesion forms witch revealed that ratios had significantly increased in lesional skin compared with non lesional one. However, at anatomy-pathology analysis, inflammatory infiltrate in psoriatic erythroderma was classical poor and no specific of this disease, such as the case of our three patients. This could explain the drop of intra-lesional inflammatory mediators.. The paradoxal low levels of proinflammatory cytokines in psoriatic erythroderma are an original and important result.

Topics: Biopsy; Dermatitis, Exfoliative; Humans; Interleukin-8; Prospective Studies; Psoriasis; Skin; Tumor Necrosis Factor-alpha

To investigate whether lithium carbonate, propranolol or chloroquine aggravate psoriasis through influencing cytokines of the psoriatic cytokine network, HaCaT keratinocytes were stimulated with TNF-a after treatment with these drugs. Protein secretion of a set of multiple different cytokines and growth factors in culture supernatants were measured by using a cytokine antibody array technology. Expression of IL-8 and IL-6 mRNA was determined by real-time PCR. In culture supernatants of TNF-alpha-stimulated HaCaT cells, production of IL-6 and TNF-alpha could be enhanced by lithium carbonate; production of IL-6 and a panel of cytokines and growth factors could be enhanced by propranolol hydrochloride; and IL-6 was up-regulated by chloroquine diphosphate as well. Real-time PCR analysis showed a significantly dose-dependent increase of IL-8 and IL-6 mRNA expression in HaCaT cells stimulated with TNF-a as compared to cells without TNF-alpha-stimulation, the mRNA expression of IL-8 was higher than that of IL-6 with the same concentration of TNF-alpha (P < 0.01). Compared with HaCaT cells cultured with medium alone, propranolol hydrochloride at the concentration of 1 x 10(-6) mol x L(-1) could stimulate HaCaT cells to express higher level of IL-6 mRNA (P < 0.05). The drugs investigated show a modulatory effect on certain cytokines and growth factors which are able to modulate inflammatory type of immune reaction present in psoriatic lesions.

Topics: Adrenergic beta-Antagonists; Antimalarials; Cells, Cultured; Chloroquine; Humans; Interleukin-6; Interleukin-8; Keratinocytes; Lithium Carbonate; Propranolol; Psoriasis; RNA, Messenger; Tumor Necrosis Factor-alpha

Toll-like receptors (TLRs) are crucial players in the innate immune response to microbial invaders. The lipophilic yeast Malassezia furfur has been implicated in the triggering of scalp lesions in psoriasis. The aim of the present study was to assess the role of TLRs in the defence against M. furfur infection. The expression of the myeloid differentiation factor 88 (MyD88) gene, which is involved in the signalling pathway of many TLRs, was also analysed. In addition, a possible correlation of antimicrobial peptides of the beta-defensin family to TLRs was tested. Human keratinocytes infected with M. furfur and a variety of M. furfur-positive psoriatic skin biopsies were analysed by RT-PCR, for TLRs, MyD88, human beta-defensin 2 (HBD-2), HBD-3 and interleukin-8 (IL-8) mRNA expression. When keratinocytes were infected with M. furfur, an up-regulation for TLR2, MyD88, HBD-2, HBD-3 and IL-8 mRNA was demonstrated, compared to the untreated cells. The same results were obtained when psoriatic skin biopsies were analysed. The M. furfur-induced increase in HBD-2 and IL-8 gene expression is inhibited by anti-TLR2 neutralising antibodies, suggesting that TLR2 is involved in the M. furfur-induced expression of these molecules. These findings suggest the importance of TLRs in skin protection against fungi and the importance of keratinocytes as a component of innate immunity.

Topics: Adaptor Proteins, Signal Transducing; beta-Defensins; Biopsy; Cell Proliferation; Cells, Cultured; Dermatomycoses; Gene Expression Regulation; Humans; Interleukin-8; Keratinocytes; Malassezia; Myeloid Differentiation Factor 88; Psoriasis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Skin; Toll-Like Receptor 2

Alterations in specific signal transduction pathways may explain the increased expression of proinflammatory cytokines seen in inflammatory diseases such as psoriasis. We reveal increased TNF-alpha protein expression, but similar TNF-alpha mRNA levels, in lesional compared with nonlesional psoriatic skin, demonstrating for the first time that TNF-alpha expression in lesional psoriatic skin is regulated posttranscriptionally. Increased levels of activated MAPK-activated protein kinase 2 (MK2) together with increased MK2 kinase activity were found in lesional compared with nonlesional psoriatic skin. Immunohistochemical analysis showed that activated MK2 was located in the basal layers of the psoriatic epidermis, whereas no positive staining was seen in nonlesional psoriatic skin. In vitro experiments demonstrated that both anisomycin and IL-1beta caused a significant activation of p38 MAPK and MK2 in cultured normal human keratinocytes. In addition, TNF-alpha protein levels were significantly up-regulated in keratinocytes stimulated with anisomycin or IL-1beta. This increase in TNF-alpha protein expression was completely blocked by the p38 inhibitor, SB202190. Transfection of cultured keratinocytes with MK2-specific small interfering RNA led to a significant decrease in MK2 expression and a subsequent significant reduction in the protein expression of the proinflammatory cytokines TNF-alpha, IL-6, and IL-8, whereas no change in the expression of the anti-inflammatory cytokine IL-10 was seen. This is the first time that MK2 expression and activity have been investigated in an inflammatory disease such as psoriasis. The results strongly suggest that increased activation of MK2 is responsible for the elevated and posttranscriptionally regulated TNF-alpha protein expression in psoriatic skin, making MK2 a potential target in the treatment of psoriasis.

Topics: Adult; Anisomycin; Cells, Cultured; Enzyme Activation; Fluorescent Antibody Technique; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Intracellular Signaling Peptides and Proteins; p38 Mitogen-Activated Protein Kinases; Protein Kinases; Protein Serine-Threonine Kinases; Psoriasis; RNA Processing, Post-Transcriptional; RNA, Messenger; Skin; Tumor Necrosis Factor-alpha; Up-Regulation

The pro-inflammatory cytokine interleukin-1 (IL-1) is constitutively expressed by keratinocytes in vivo and has been shown to be expressed in psoriatic lesional skin. To determine what role the IL-1 system might contribute to the inflammatory process in psoriasis, semi-quantitative RT-PCR and cRNA microarray studies were performed on biopsies excised from lesional and non-lesional skin. Whilst IL-1alpha mRNA levels showed a reduction in lesional skin in a subset of patients, steady state IL-1beta mRNA was increased markedly. Neither of the two IL-1 receptor transcripts nor total IL-1 receptor antagonist exhibited major changes within the lesion. Expression of the IL-1-induced chemokine IL-8 was only observed in lesional epidermis. Functional genomic experiments comparing transcriptome profiles derived from psoriatic lesional skin and IL-1 stimulated keratinocytes demonstrated a striking level of overlap. Taken together, these data suggest that IL-1 is likely to be an important mediator in the initiation and maintenance of psoriatic plaques and may represent an attractive therapeutic target.

Topics: Adult; Aged; Aged, 80 and over; Cells, Cultured; Epidermal Cells; Female; Humans; Immunoenzyme Techniques; Inflammation Mediators; Interleukin-1; Interleukin-8; Male; Middle Aged; Psoriasis; RNA, Messenger; Skin

Mitogen- and stress-activated protein kinase 1 (MSK1) is a downstream target of both the p38 and extracellular signal-regulated kinase 1/2 (ERK1/2) mitogen-activated protein kinases (MAPKs). MSK1 stimulates transcription of different pro-inflammatory genes through activation of transcription factors. The purpose of this study was to investigate the expression and activation of MSK1 in lesional psoriatic skin and its role in cytokine production in cultured normal human keratinocytes. Western blotting revealed a consistent and significant increase in phosphorylated (activated) MSK1(Ser376) in lesional psoriatic skin. Immunofluorescence staining revealed the phosphorylated MSK1(Thr581) to be localized in the basal layers of the epidermis in lesional psoriatic skin. No staining was found in non-lesional psoriatic skin. Cultured human keratinocytes incubated with anisomycin or IL-1beta resulted in the phosphorylation of the p38 MAPK and MSK1(Ser376). MSK1(Ser376) phosphorylation was inhibited by pre-incubation with the p38 inhibitor SB 202190. Transfection of the keratinocytes with specific MSK1 small interfering RNA resulted in 80% reduction of MSK1 expression and 51, 40, and 31% decrease in IL-6, IL-8, and tumor necrosis factor-alpha protein production, respectively. This study demonstrates for the first time the expression of MSK1 in epidermal keratinocytes and increased activation focally in psoriatic epidermis. As MSK1 regulates the production of pro-inflammatory cytokines, it may play a role in the pathogenesis of psoriasis.

Topics: Adult; Anisomycin; Cells, Cultured; Cyclic AMP Response Element-Binding Protein; Cytokines; DNA-Binding Proteins; Enzyme Activation; Enzyme Inhibitors; Epidermal Cells; Epidermis; Fluorescent Antibody Technique; Humans; Imidazoles; Interleukin-1; Interleukin-6; Interleukin-8; Keratinocytes; Nuclear Proteins; Protein Synthesis Inhibitors; Psoriasis; Pyridines; Regulatory Factor X Transcription Factors; Ribosomal Protein S6 Kinases, 90-kDa; RNA, Small Interfering; Transcription Factors; Tumor Necrosis Factor-alpha

Fibroblasts have been implicated in psoriatic inflammatory processes. The aim of the study was to evaluate soluble interleukin 2 receptor (sIL-2R), interleukin 6 (IL-6), and interleukin 8 (IL-8) plasma levels in psoriatic patients and IL-6 and IL-8 levels in fibroblast culture supernatants. Cytokines levels in plasma and supernatants were measured by ELISA. Plasma sIL-2R, IL-6, and IL-8 levels were higher before the treatment in comparison to healthy controls (P < 0.001) and decreased after treatment. Fibroblasts from healthy controls, psoriatic lesional skin, and noninvolved psoriatic skin, when stimulated with tumor necrosis factor alpha, released considerable amounts of IL-6 and IL-8. No significant difference between healthy controls and psoriatic fibroblasts was observed. Monitoring plasma sIL-2R levels could be employed as a reliable method of psoriasis activity. IL-8 and IL-6 plasma levels seem to reflect psoriasis activity, and treatment response, respectively. Fibroblasts are not a major source of increased IL-6 and IL-8 production in psoriasis.

Topics: Adult; Aged; Enzyme-Linked Immunosorbent Assay; Female; Fibroblasts; Gene Expression Regulation; Humans; Inflammation; Interleukin-6; Interleukin-8; Male; Middle Aged; Psoriasis; Receptors, Interleukin-2

Mast cells are involved in many disorders where the triggering mechanism that leads to degranulation and/or cytokine secretion has not been defined. Several chronic inflammatory diseases are associated with increased mast cell numbers and upregulation of the TNF receptor family member CD30, but the role of elevated CD30 expression is poorly understood. Here we report what we believe to be a novel way to activate mast cells with CD30 that leads to degranulation-independent secretion of chemokines. CD30 induced a de novo synthesis and secretion of the chemokines IL-8, macrophage inflammatory protein-1alpha (MIP-1alpha), and MIP-1beta, a process involving the MAPK/ERK pathway. Mast cells were found to be the predominant CD30 ligand-positive (CD30L-positive) cell in the chronic inflammatory skin diseases psoriasis and atopic dermatitis, and both CD30 and CD30L expression were upregulated in lesional skin in these conditions. Furthermore, the number of IL-8-positive mast cells was elevated both in psoriatic and atopic dermatitis lesional skin as well as in ex vivo CD30-treated healthy skin organ cultures. In summary, characterization of CD30 activation of mast cells has uncovered an IgE-independent pathway that is of importance in understanding the entirety of the role of mast cells in diseases associated with mast cells and CD30 expression. These diseases include Hodgkin lymphoma, atopic dermatitis, and psoriasis.

Topics: Adolescent; Adult; CD30 Ligand; Cell Degranulation; Cells, Cultured; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokines; Cytokines; Dermatitis, Atopic; Female; Fetal Blood; Gene Expression; Histamine Release; Humans; Interleukin-8; Ki-1 Antigen; Leukotrienes; Macrophage Inflammatory Proteins; Male; MAP Kinase Signaling System; Mast Cells; Middle Aged; Psoriasis; Skin Diseases; Tryptases; Up-Regulation

Avarol, a marine sesquiterpenoid hydroquinone, and 14 avarol derivatives have shown interesting anti-inflammatory properties in previous studies. In this study, avarol and derivatives were evaluated in high-throughput keratinocyte culture models using cytokeratin 10 and SKALP/Elafin expression as markers for respectively normal and psoriatic differentiation. Avarol and five of its derivatives (5, 10, 13, 14 and 15) were selected for further study. Only 10, 13, 14 and 15 were able to inhibit keratinocyte cell growth. Changes in expression levels of 22 genes were assessed by quantitative real time PCR (qPCR). From these genes, TNFalpha mRNA levels showed the strongest changes. For compound 13, 15 and dithranol (used as a model antipsoriatic drug), a dose-dependent downregulation of TNFalpha mRNA was found. The changes in TNFalpha mRNA were confirmed at the protein level for compound 13. Additionally, this compound was able to reduce also IL-8 and COX-2 mRNA levels and this effect was correlated with a reduction in COX-2 protein expression. The mechanism of action of this compound involves at least the inhibition of NF-kappaB-DNA binding activity. In conclusion, our high-throughput screening models in combination with quantitative assessment of changes in gene expression profiles identified the avarol derivative 13, a benzylamine derivative of avarol at the 4' position of benzoquinone ring, as an interesting anti-psoriatic drug candidate that inhibits keratinocyte cell growth and TNFalpha and COX-2 expression.

Topics: Animals; Antineoplastic Agents; Cell Differentiation; Cell Proliferation; Cells, Cultured; Cyclooxygenase 2; Drug Evaluation, Preclinical; Dysidea; Elafin; Enzyme-Linked Immunosorbent Assay; Humans; In Vitro Techniques; Interleukin-8; Keratinocytes; Keratins; Membrane Proteins; Psoriasis; RNA, Messenger; Sesquiterpenes; Tumor Necrosis Factor-alpha

Topics: Case-Control Studies; Cytokines; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Psoriasis; Receptors, Interleukin-2; Saudi Arabia; Tumor Necrosis Factor-alpha

To screen human scFv against IL-8 from phage antibody library.. IL-8-His fusion protein was expressed in E.coli BL21 (DE3) transfected with prokaryotic expression vector pRSET-IL-8 and was purified by affinity chromatography. Specific antibody was screened by 3 rounds of panning of phage antibody library with the fusion protein. The antigen binding activity and DNA sequences of positive clones were determined and analyzed.. After 3 rounds of panning, 2 positive clones were obtained which could bind IL-8 specifically. DNA sequence analysis showed both of the 2 VH genes belonged to human IgG VH3 subgroup, and the Vlambda gene belonged to human VlambdaDPL5 and VlambdaDPL2 subgroups,respectively.. It is feasible to obtain human anti-IL-8 antibody by phage display technique, which provides the basis for further research on psoriasis and other related diseases.

Topics: Antibodies; Bacteriophages; Base Sequence; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Humans; Immunoglobulin Heavy Chains; Immunoglobulin Variable Region; Interleukin-8; Molecular Sequence Data; Peptide Library; Psoriasis; Recombinant Fusion Proteins

Tetradecylthioacetic acid (TTA) is a hypolipidemic antioxidant with immunomodulating properties involving activation of peroxisome proliferator-activated receptors (PPARs). Human endothelial cells express PPARs. We hypothesized that TTA could modulate endothelial cell activation at least partly through PPAR-related mechanisms.. We explored this hypothesis by different experimental approaches involving both in vitro studies in human endothelial cells (HUVECs) and in vivo studies in humans and PPAR-alpha-/- mice. Our main findings were as follows: (1) TTA suppressed the tumor necrosis factor alpha-induced expression of vascular cell adhesion molecule 1 (VCAM-1) and interleukin 8 (IL-8) in HUVECs. (2) No TTA-mediated attenuation of VCAM-1 and chemokine expression was seen in the liver of PPAR-alpha-/- mice. (3) Whereas TTA markedly enhanced PPAR-alpha-target genes in the liver of wild-type, but not of PPAR-alpha-/-, mice, no such effect on PPAR-alpha-target genes was seen in HUVECs. (4) The relevance of our findings to human disease was suggested by a TTA-mediated downregulation of serum levels of soluble VCAM-1 and IL-8 in psoriasis patients.. We show that TTA has the ability to attenuate tumor necrosis factor alpha-mediated endothelial cell activation, further supporting antiinflammatory effects of this fatty acid, possibly involving both PPAR-alpha-dependent and -independent pathways.

Topics: Adult; Aged; Animals; Anti-Inflammatory Agents; Antioxidants; Cell Adhesion; Cells, Cultured; Chemokine CCL2; Endothelium, Vascular; Female; Gene Expression; Humans; Interleukin-8; Male; Mice; Mice, Inbred Strains; Mice, Mutant Strains; Middle Aged; Monocytes; Neutrophils; PPAR alpha; PPAR delta; Psoriasis; Sulfides; Tumor Necrosis Factor-alpha; Umbilical Veins; Vascular Cell Adhesion Molecule-1

Nuclear factor-kappaB (NF-kappaB) is an inducible nuclear transcription factor regulating a range of cellular processes. An imbalance of the DNA binding activity of NF-kappaB may, therefore, be part of the pathophysiological mechanisms in psoriasis. The purpose of this study was to determine the NF-kappaB DNA binding activity in psoriatic skin using three different kappaB sites and to determine how DNA binding activity was modulated by the anti-psoriatic drug calcipotriol. By electrophoretic mobility shift assay, we demonstrated that the NF-kappaB DNA binding to the p53 kappaB site was decreased, whereas the NF-kappaB DNA binding to the interleukin-8 (IL-8) kappaB site was increased in lesional psoriatic skin compared with non-lesional psoriatic skin. No regulation was seen on the NF-kappaB DNA binding to the major histocompatibility complex class I kappaB site. These changes were paralleled by a similar decrease in p53 expression and an increase in IL-8 expression in involved psoriatic skin compared with uninvolved skin as determined by quantitative RT-PCR. The alteration in NF-kappaB DNA binding activity was neither accompanied by any change in the expression of the inhibitor kappaB (IkappaB) kinases, IKKalpha, IKKbeta, and IKKgamma nor in the expression of the NF-kappaB inhibitor proteins, IkappaBalpha and IkappaBbeta. Immunofluorescence analysis revealed that p65 was sequestered in the cytoplasm of keratinocytes, whereas p50 exhibited a cytoplasmic as well as a nuclear localization. Interestingly, this distribution of p50 and p65 was similar in lesional and non-lesional psoriatic skin. Topical application of calcipotriol to lesional psoriatic skin for 4 d resulted in increased NF-kappaB binding to the p53 kappaB site and decreased NF-kappaB binding to the IL-8 kappaB site. Taken together, our data demonstrate that the NF-kappaB DNA binding activity is regulated in a specific manner in psoriatic skin depending on the kappaB sites investigated, and that topical treatment of psoriatic skin normalizes the abnormal NF-kappaB binding activity seen in lesional psoriatic skin.

Topics: Administration, Cutaneous; Adult; Blotting, Western; Calcitriol; Cells, Cultured; Dermatologic Agents; DNA; Electrophoretic Mobility Shift Assay; Fluorescent Antibody Technique; Gene Expression Regulation; Humans; I-kappa B Kinase; I-kappa B Proteins; Interleukin-8; Keratinocytes; NF-kappa B; Protein Isoforms; Protein Serine-Threonine Kinases; Psoriasis; Response Elements; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Skin; Tumor Suppressor Protein p53

Munro's microabscesses are a characteristic histopathologic feature of psoriasis vulgaris; however, the pathomechanisms underlying the migration of transepidermal leukocytes (PMNs) have not been fully elucidated yet.. Since the lesional scale extracts contain potent chemoattractants, such as IL-8 and C5a fragments, we studied their location in the lesions of psoriasis vulgaris and PPP with immunohistochemical techniques.. Localization of IL-8 was not detected in the subcorneal keratinocytes but was demonstrated only in the basal keratinocytes together with migrating PMNs. In contrast, the presence of a complement fragment, C3b, was observed on the cell membranes of subcorneal keratinocytes, suggesting that these were the sites of complement activation.. Such distinct localization of IL-8 and complement components suggests that the intraepidermal migration of PMNs takes place first according to the concentration gradient of IL-8, and thereafter they are guided by complement components to the final destination, the subcorneal portion of the lesional skin.

Topics: Cell Movement; Chemotactic Factors; Complement C5a; Humans; Interleukin-8; Keratinocytes; Neutrophils; Psoriasis; Skin

Previous studies of acute generalized exanthematous pustulosis, a peculiar drug hypersensitivity reaction, suggested that CXCL8-producing T cells regulate sterile, polymorphonuclear neutrophil-rich skin inflammations. In this study, we test the hypothesis of whether CXCL8-producing T cells are present in autoinflammatory diseases like pustular psoriasis and Behçet's disease. Immunohistochemistry of normal skin revealed few CD4+ and CD8+ T cells, few CXCL8+ cells, and no neutrophilic infiltration, whereas in acute exacerbations of atopic dermatitis, numerous CD4+ T cells but few CD8+ T cells, neutrophils, or CXCL8+ cells were detected. In contrast, a pronounced infiltration of neutrophils and of predominantly CD4+ T cells was observed in skin biopsies from pustular psoriasis, Behçet's disease, and acute generalized exanthematous pustulosis, with infiltrating T cells strongly positive for CXCL8 and the chemokine receptor CCR6. Skin-derived T cell clones from pustular skin reactions were positive for CCR6 but negative for CCR8 and secreted high amounts of CXCL8 and GM-CSF, often together with IFN-gamma and TNF-alpha after in vitro stimulation. Moreover, some skin-derived T cell clones from Behçet's disease and from pustular psoriasis predominantly produced CXCL8 and GM-CSF, but failed to secrete IL-5 and IFN-gamma. These cells might represent a particular subset as they differ from both Th1 as well as Th2 T cells and are associated with a unique, neutrophil-rich sterile inflammation. Our findings suggest that CXCL8/GM-CSF-producing T cells may orchestrate neutrophil-rich pathologies of chronic autoinflammatory diseases like pustular psoriasis and Behçet's disease.

Topics: Adult; Aged; Behcet Syndrome; CD4 Antigens; CD8 Antigens; Chemokine CCL20; Chemokines, CC; Dermatitis, Atopic; Female; Flow Cytometry; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunohistochemistry; Inflammation; Interferon-gamma; Interleukin-8; Macrophage Inflammatory Proteins; Male; Middle Aged; Neutrophils; Psoriasis; Receptors, CCR6; Receptors, Chemokine; Skin; Skin Diseases; T-Lymphocytes; Tumor Necrosis Factor-alpha

Psoriasis vulgaris, a type-1 cytokine-mediated chronic skin disease, can be treated successfully with fumaric acid esters (FAE). Beneficial effects of this medication coincided with decreased production of IFN-gamma. Since dendritic cells (DC) regulate the differentiation of T helper (Th) cells, this study focussed on effects of monomethylfumarate (MMF, bioactive metabolite of FAE) on polarization of monocyte-derived DC. MMF-incubated, lipo-polysaccharide-stimulated DC (MMF-DC) produced dramatically (p<0.05) reduced levels of IL-12p70 and IL-10 (8+/-4% and 20+/-4%, respectively) compared to control DC. MMF-DC were mature. MMF affected polarization of DC irrespective of polarization factor(s) and ligands for the various Toll-like receptors used. Coculture of MMF-DC with naive and primed allogenous Th cells resulted in lymphocytes producing less IFN-gamma, i.e. 59% and 54% of that by the respective Th cells cocultured with control DC. IL-4 production by primed, but not naive Th cells cocultured with MMF-DC was decreased as compared to cocultures with control DC. IL-10 production by naive and primed Th cells cocultured with MMF-DC and control DC did not differ. In addition, MMF inhibited LPS-induced NF-kappaB activation in DC. Together, beneficial effects of FAE in psoriasis involve modulation of DC polarization by MMF such that these cells down-regulate IFN-gamma production by Th cells.

Topics: Antigens, CD; Cell Differentiation; Cell Polarity; Coculture Techniques; Dendritic Cells; Down-Regulation; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Fumarates; Humans; Interleukin-10; Interleukin-12; Interleukin-8; Lymphocyte Activation; Maleates; Monocytes; Protein Subunits; Psoriasis; Th1 Cells; Tumor Necrosis Factor-alpha

Psoriasis is a chronic inflammatory skin disorder characterized by accumulation of Th1-type T cells and neutrophils, regenerative keratinocyte proliferation and differentiation, and enhanced epidermal production of antimicrobial peptides. The underlying cause is unknown, but there are some similarities with the immunologic defense program against bacteria. Development of psoriasiform skin lesions has been reported after administration of granulocyte colony-stimulating factor (G-CSF), a cytokine induced in monocytes by bacterial antigens. To further investigate the relation between this type of cytokine-induced dermatitis and psoriasis, we analyzed the cutaneous cytokine profile [tumor necrosis factor-alpha (TNF-alpha), interferon-gamma, transforming growth factor-beta1 (TGF-beta1), interleukin-10 (IL-10), IL-12p35 and p40, and IL-8] and expression of markers of epidermal activation [Ki-67, cytokeratin-16, major histocompatibility complex (MHC) class II, intercellular adhesion molecule-1 (ICAM-1)] in a patient who developed G-CSF-induced psoriasiform dermatitis by using quantitative real-time reverse transcriptase-polymerase chain reaction and immunohistology. The histologic picture resembled psoriasis with regard to epidermal hyperparakeratosis and the accumulation of lymphocytes in the upper corium. CD8(+) T cells were found to infiltrate the epidermis which was associated with an aberrant expression of Ki-67, cytokeratin-16, MHC class II, and ICAM-1 on adjacent keratinocytes. As compared to normal skin (n = 7), there was an increased expression of TNF-alpha, IL-12p40, and IL-8, a decreased expression of TGF-beta1, and a lack of IL-10, similar to the findings in active psoriasis (n = 8). Therefore, G-CSF may cause a lymphocytic dermatitis that, similar to psoriasis, is characterized by a pro-inflammatory Th1-type cytokine milieu and an epidermal phenotype indicative of aberrant maturation and acquisition of non-professional immune functions.

Topics: Cytokines; Drug Eruptions; Epidermis; Gene Expression; Granulocyte Colony-Stimulating Factor; Humans; Interleukin-10; Interleukin-12; Interleukin-12 Subunit p35; Interleukin-12 Subunit p40; Interleukin-8; Keratinocytes; Male; Middle Aged; Protein Subunits; Psoriasis; T-Lymphocytes; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

In this study, the phenotype of psoriatic keratinocytes and fibroblasts in reconstructed skin models was compared to those constructed from normal cells. Characterization of this model by immunohistochemistry showed that classical markers of keratinocyte differentiation exhibited similar patterns of distribution in the psoriatic models to those derived from normal cells and generally reflected in vivo observations. Some crucial differences, however, were observed between normal and psoriatic models when pro-inflammatory gene expression and keratinocyte proliferation were investigated. Notably, the chemokine receptor CXCR2 was overexpressed in the psoriatic models, and, moreover, was localized to the granular layer of keratinocytes as seen in psoriasis in vivo. Pro-inflammatory genes (tumor necrosis factor alpha [TNF-alpha], interferon gamma [IFN-gamma], and interleukin 8 [IL-8]) were expressed at high levels in the psoriatic models, but were only minimally expressed in the normal models. Models derived from uninvolved psoriatic skin showed the same gene expression profile as those derived from involved skin along with an increased proliferation rate when compared to normal models. These results suggest that psoriatic individuals possess an inherent predisposition to develop the disease phenotype even in the absence of T cells. This study represents a comprehensive characterization of psoriatic human skin reconstructed in vitro, and demonstrates the potential of this model as a valuable tool in drug discovery.

Topics: Adult; Aged; Antibody Specificity; Cells, Cultured; Chemokine CXCL1; Chemokines, CXC; Female; Gene Expression; Humans; Immunohistochemistry; Intercellular Signaling Peptides and Proteins; Interferon-gamma; Interleukin-6; Interleukin-8; Keratinocytes; Male; Middle Aged; Organ Culture Techniques; Phenotype; Psoriasis; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Tumor Necrosis Factor-alpha

Psoriatic plaques have been shown to contain increased levels of proinflammatory cytokines. Serum levels of interleukin (IL)-6, IL-7, IL-8, and interferon (IFN)-gamma have been reported elevated in psoriatic patients.. To evaluate serum cytokine profiles in psoriasis patients by improved enzyme-linked immunosorbent assay (ELISA) technique and to correlate these levels with disease severity.. We analyzed single serum samples from 10 patients with active untreated psoriasis, two patients with active treated psoriasis, and five healthy volunteers for major T helper type 1 and T helper type 2 cytokines using the LINCOplex ELISA multi-analyte detection system that permits simultaneous detection of multiple cytokines from a single sample. The disease severity, including erythema, induration, scale, and surface area, was assessed.. IFN-gamma was markedly elevated in all sera from psoriasis patients, 33.8 +/- 1.3 pg/ml (mean +/- standard error) versus 8 +/- 1.5 pg/ml for normal controls (p < 0.01), and positively correlated with all indices of disease severity (Spearman r > 0.6). IL-8 was also increased in psoriasis patients (24.4 +/- 1.8 pg/ml) versus normal controls (3.6 +/- 1.2 pg/ml) (p < 0.05) and positively correlated with the degree of erythema (Spearman r > 0.6). Mean IL-12 levels were decreased in sera from psoriasis patients (8.5 +/- 1.2 pg/ml) compared with normal controls (42.2 +/- 5.3 pg/ml) (p < 0.01). Also, serum IL-10 levels were below detection levels in psoriatics compared with controls (6.4 +/- 1.3 pg/ml).. This new ELISA system allowed rapid and reliable detection of numerous cytokines in single serum samples from patients with psoriasis. We observed that IFN-gamma and IL-8 cytokines were elevated in psoriatics and correlated with parameters of disease severity while IL-10 and IL-12 were decreased.

Topics: Adolescent; Adult; Aged; Biomarkers; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-8; Male; Middle Aged; Psoriasis; Severity of Illness Index; Th1 Cells; Th2 Cells

Chemokines represent a family of potent biological mediators. Within the group of receptors mediating their effects, a promiscuous receptor has been found which is able to bind and inactivate diverse chemokines of both C-C and C-X-C families. It is co-localized with blood group antigens of the Duffy system on the same glycoprotein and expressed on red blood cells as well as post-capillary blood vessels. In the present study three aspects of Duffy pathophysiology were studied: firstly the amount of IL-8 and RANTES binding to red blood cells and its correlation to disease activity of psoriatic patients, secondly the distribution of Duffy phenotype among psoriatic patients and thirdly the expression of Duffy antigen in normal vs psoriatic skin. Red blood cells from psoriatic patients (n=50) were lysed by triton X (1%) and supernatants tested in IL-8- and RANTES sandwich-ELISA. Duffy phenotype of psoriatic patients (n=50) was assessed by typing red blood cells with specific antisera in indirect Coombs technique. For immunohistochemical detection in normal and psoriatic skin (n=10 respectively) a specific monoclonal antibody (Fy6) was used. Neither IL-8- nor RANTES-levels on red blood cells correlated to disease activity and distribution of Duffy phenotype in psoriatics was not significantly altered when compared to the normal population. Furthermore, Duffy antigen was expressed in a similar pattern in normal and psoriatic skin at all parts of vasculature, albeit much more abundantly in diseased skin. Altogether, chemokine binding to red blood cells seems of minor importance in psoriasis. However, Duffy antigen together with other binding mechanisms like proteoglycans may play a role at local level by binding locally produced chemokines. Thus biological effects of chemokines are both restricted and focussed to dermal tissue.

Topics: Adult; Aged; Aged, 80 and over; Chemokine CCL5; Dose-Response Relationship, Drug; Duffy Blood-Group System; Enzyme-Linked Immunosorbent Assay; Erythrocytes; Female; Humans; Immunohistochemistry; Interleukin-8; Male; Middle Aged; Phenotype; Protein Binding; Protein Structure, Tertiary; Psoriasis; Skin

Leopoldine spa water is a hypotonic water rich in sulphate that has been used occasionally for balneological treatments in psoriatics. We evaluated the anti-inflammatory effects of this salso-sulphate water on the skin of subjects with psoriasis.. We selected 10 volunteer subjects (23-58 years old), who presented symmetrical, bilateral psoriasis involving at least 40% of the body surface. All the subjects were subjected to the following treatment schedule: (i) immersion of both arms in water twice a day [the right arm was immersed in Leopoldine spa water at its natural source temperature (27.2 degrees C) for 30 min, and the left arm was immersed in double-distilled water for 30 min at a constant temperature of 27 degrees C]; (ii) both arms were exposed to the sun for 60 min after each immersion; and (iii) vaseline containing moisturizing creams were applied liberally. The treatment was continued for 4 weeks and was well tolerated by all subjects. Response to treatment was assessed by means of the modified Psoriasis Area and Severity Index (PASI). Six of the 10 subjects volunteered to undergo a cutaneous biopsy of lesional skin both before and 4 weeks after treatment to allow for assessment of modifications of the cutaneous infiltrate in the areas treated.. At the end of 4 weeks the average pretreatment PASI score of the left arms was 5.72 (range 4-9.6) while the right arms had a mean PASI of 5.56 (range of 3.2-9). At the end of the study the average PASI score was 0.78 for the arms treated with Leopoldine spa water and 2.83 for the arms treated with double-distilled water. The mean PASI improvement score for the Leopoldine spa water treated arms was 85.9% while the double-distilled water treated arms showed a PASI improvement score of 50.5%. An immunohistological study showed significant differences between the cutaneous samples taken 4 weeks after treatment and those taken before treatment with Leopoldine spa water. There were significant decreases in the numbers of epidermal CD4+ and CD8+ T lymphocytes and CD1a+ Langerhans cells (microscopic field at x 22 objective), as well as a decrease in the epidermal keratinocyte expression of intercellular adhesion molecule-1 and interleukin-8 and the dermal expression of CD4+ and CD8+ T lymphocytes.. These data show the potential anti-inflammatory effects of Leopoldine mineral waters on human skin affected by psoriasis.

Topics: Adult; Balneology; Female; Humans; Immunoenzyme Techniques; Intercellular Adhesion Molecule-1; Interleukin-8; Italy; Male; Middle Aged; Mineral Waters; Psoriasis; Severity of Illness Index; Treatment Outcome

To explore the therapeutic mechanism of Xiaoyin Jiedu Yin (XYJDY) in treating psoriasis.. Abnormal elevation of interleukin 8 (IL-8) level in mice was induced by Staphylococcus aureus enterotoxin B (SEB), then the antagonizing effects of XYJDY and Composite Qingdai Capsule were observed.. Serum IL-8 level arose and reached the peak 3-5 hrs after SEB induction. XYJDY could antagonize the induction significantly, and the effect was more significant when large dosage was given.. XYJDY, in either large or small dose, could effectively antagonize the SEB induction on IL-8. Applying with superantigen theory, this fact could be used to elucidate the mechanism of XYJDY in removing Heat, cooling blood and detoxifying at cell biologic level.

Topics: Animals; Drugs, Chinese Herbal; Enterotoxins; Interleukin-8; Male; Mice; Phytotherapy; Psoriasis; Staphylococcus aureus

Atopic dermatitis (AD) and psoriasis are genetically determined inflammatory skin disorders. Keratinocytes actively participate in cutaneous inflammatory responses by elaborating various chemokines.. We investigated the capacity of IL-4, IFN-gamma, and TNF-alpha to modulate the expression of CCL and CXCL chemokines in cultured keratinocytes from patients and healthy individuals, as well as chemokine expression in situ.. Keratinocyte cultures were established from normal-looking skin of adult patients with AD or psoriasis vulgaris and from healthy subjects. Monocyte chemoattractant protein 1 (MCP-1)/CCL2, RANTES/CCL5, IL-8/CXCL8, and IFN-gamma-induced protein of 10 kd (IP-10)/CXCL10 production was evaluated at the mRNA and protein levels by using RNase protection assay and ELISA, respectively. The expression of the same chemokines was studied in chronic lesional skin by means of immunohistochemistry or in situ hybridization.. Only IL-8 mRNA was detected in unstimulated ke-ratinocyte cultures. MCP-1 and IP-10 were potently induced by IFN-gamma, whereas IL-8 and RANTES were preferentially upregulated by TNF-alpha and, to a lesser extent, by IFN-gamma. IL-4 weakly induced IP-10, RANTES, and IL-8 but not MCP-1. Keratinocytes of patients with AD invariably responded with significantly earlier and higher RANTES expression. By contrast, keratinocytes of patients with psoriasis displayed much higher levels of both constitutive and induced IL-8 and a stronger induction of MCP-1 and IP-10. RANTES and MCP-1 mRNA(+) keratinocytes were detected in the basal layer of lesions of patients with AD and psoriasis. IP-10 and IL-8 were consistently upregulated in the epidermis of patients with psoriasis but not in lesions of patients with AD.. Keratinocytes of patients with AD and psoriasis show an intrinsically abnormal and different chemokine production profile and may thus favor the recruitment of distinct leukocyte subsets into the skin.

Topics: Adult; Autoantigens; Cells, Cultured; Chemokine CCL2; Chemokine CCL5; Chemokine CXCL10; Chemokines; Chemokines, CXC; Chemotaxis, Leukocyte; Cytokines; Dermatitis, Atopic; Female; Gene Expression Regulation; Humans; Interferon-gamma; Interleukin-4; Interleukin-8; Keratinocytes; Leukocytes; Male; Middle Aged; Psoriasis; T-Lymphocyte Subsets; Tetradecanoylphorbol Acetate; Tumor Necrosis Factor-alpha

Psoriasis is a disease marked by keratinocyte hyperproliferation, neutrophil and lymphocyte infiltration, and aberrant epidermal and dermal expression of cytokines. Previously, it has been shown that LIF appears to be involved in skin inflammation and can induce the expression of IL-8. We sought to determine whether expression of LIF is abnormal in lesional psoriatic skin and whether this correlates with the expression of IL-8. Using reverse transcription polymerase chain reaction, we measured the expression of LIF and IL-8 mRNA in biopsies from normal individuals, non-lesional psoriatic skin, and lesional psoriatic skin. No difference was seen between the expression of IL-8 and LIF in normal and in non-lesional psoriatic skin. However, LIF expression in lesional skin was increased 160% compared with normal biopsies or non-lesional skin (p < 0.001). Immunostaining of frozen sections showed that the expression of LIF protein was principally suprabasal and, in the majority of sections, concentrated mainly in the stratum corneum of the lesional skin, whereas it was mainly in the stratum spinosum of the normal/non-lesional skin. IL-8 mRNA expression did not differ between the non-lesional and normal skin, but expression in the lesional skin was 17.6-fold greater than in normal skin (p < 0.001), and this expression was correlated with the increased LIF expression (r = 0.67, p < 0.001). Although a significant negative correlation was demonstrated between LIF mRNA expression and the duration of the last outbreak of the disease, no other correlations were found between levels of cytokine expression and a variety of parameters including PASI score. These data suggest a role for keratinocyte LIF in the psoriatic lesion and a link between LIF and IL-8 expression.

Topics: Adult; Aged; Case-Control Studies; Female; Growth Inhibitors; Humans; Immunohistochemistry; Interleukin-6; Interleukin-8; Leukemia Inhibitory Factor; Lymphokines; Male; Middle Aged; Psoriasis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

We performed an immunohistochemical study to try to determine the cellular source of interleukin-8 (IL-8) in psoriatic skin lesions. IL-8 was positively stained in the vast majority of neutrophils but not in the mononuclear cells, macrophages, or keratinocytes. IL-8-positive neutrophils were seen both in Munro's microabcesses in cases of psoriasis vulgaris and in a small spongiform pustule and much larger macropustules of Kogoj in cases of pustular psoriasis. Some IL-8-positive neutrophils were observed in the upper dermis of pustular psoriasis. The staining was considered to be specific because it could be completely blocked by preabsorption with recombinant IL-8. In addition, stimulation of human neutrophils with lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-alpha) for 18 h induced IL-8 production in vitro. In our study, IL-8 was expressed in the neutrophils of psoriasis, suggesting that neutrophils are one of the sources of IL-8 in psoriasis. The expression of IL-8 and the influx of neutrophils led us to speculate that the IL-8 autocrine and/or paracrine system functions in the formation of the microabcesses and pustules in proriasis.

Topics: Humans; Immunohistochemistry; In Vitro Techniques; Interleukin-8; Lipopolysaccharides; Neutrophils; Psoriasis; Tumor Necrosis Factor-alpha

An important cellular aberration at sites of psoriatic inflammation is an increase in the number of dermal mast cells. Being multifactorial immune effector cells, it is believed that mast cells play an essential role in perpetuating the inflammatory process of psoriasis. However, factors responsible for the infiltration and accumulation of mast cells in psoriatic lesions are largely unknown. Recent studies have demonstrated that Interleukin-8 (IL-8) exerts strong chemotactic effects on mast cells in vitro. Overexpression of IL-8 has also been reported in psoriatic lesions. In this study, we have found a correlation between the expression of IL-8 and dermal mast cell density in lesional psoriatic skin as compared to nonlesional psoriatic skin.. Four-mm punch biopsies were taken from 14 psoriatic patients and eight healthy volunteers. Using immunohistochemical techniques, 8 microm sections of lesional psoriatic, nonlesional psoriatic, and normal control samples were evaluated for dermal mast cell density and the density of IL-8 expressing keratinocytes.. It was found that dermal mast cell density in lesional psoriatic, nonlesional psoriatic, and normal skin was 105.4 +/- 71.2, 42.3 +/- 30.1, and 47.5 +/- 32.5 mast cells/mm(2), respectively. IL-8+ keratinocyte density in lesional psoriatic, non lesional psoriatic, and normal skin was 171.5 +/- 67.1, 25.4 +/- 14.9 and 20.6 +/- 8.7 IL-8+ Keratinocytes/mm(2), respectively.. The results of this study suggest that increased levels of IL-8 in the keratinocytes of psoriatic plaques play a contributing role in the migration of mast cells to lesion sites.

Topics: Biopsy, Needle; Culture Techniques; Female; Humans; Immunohistochemistry; Interleukin-8; Keratinocytes; Male; Mast Cells; Psoriasis; Reference Values; Sensitivity and Specificity; Skin

Keratinocytes (KC) are important source of and targets for several cytokines. Although KC express IL-15 mRNA, the functional effects of IL-15 on these epithelial cells remain to be dissected. Investigating primary human foreskin KC and HaCaT cells, we show here by semiquantitative RT-PCR and flow cytometric analysis that both translate IL-15 and IL-15R mRNA and express IL-15 and IL-15Ralpha protein on the cell surface, suggesting that human KC can employ IL-15 for juxtacrine signaling. While IL-15 exerted no significant effect on KC proliferation and IL-6 or IL-8 secretion, IL-15 inhibited both anti-Fas and methylcellulose-induced KC apoptosis in vitro. This is in line with the recognized potent anti-apoptotic effects of IL-15. IL-2, whose receptor shares two components with the IL-15R, failed to inhibit KC apoptosis. Together with the role of IL-15 in sustaining chronic immune reactions, this invited the question of whether a reduction of KC apoptosis by IL-15 may be involved in the pathogenesis of psoriasis, a chronic hyperproliferative inflammatory skin disease characterized by abnormally low KC apoptosis in the epidermis. Remarkably, compared with nonlesional psoriatic skin and skin of healthy volunteers, lesional psoriatic epidermis showed high IL-15 protein expression in the epidermis and enhanced binding activity for IL-15. Therefore, antagonizing the inhibitory effects of IL-15 on KC apoptosis deserves exploration as a novel therapeutic strategy in psoriasis management.

Topics: Apoptosis; Binding Sites; Cell Division; Cell Membrane; Cells, Cultured; Epidermis; fas Receptor; Gene Expression Regulation; Humans; Immune Sera; Immunosuppressive Agents; Interleukin-15; Interleukin-6; Interleukin-8; Keratinocytes; Methylcellulose; Psoriasis; Receptors, Interleukin-15; Receptors, Interleukin-2; RNA, Messenger

The analysis of cytokine profiles plays a central part in the characterization of disease-related inflammatory pathways and the identification of functional properties of immune cell subpopulations. Because tissue biopsy samples are too small to allow the detection of cytokine protein, the detection of mRNA by RT-PCR analysis is often used to investigate the cytokine milieu in inflammatory lesions. RT-PCR itself is a qualitative method, indicating the presence or absence of specific transcripts. With the use of internal or external standards it may also serve as a quantitative method. The most widely accepted method is quantitative competitive RT-PCR, based on internal shortened standards. Recently, online real-time PCR has been introduced (LightCycler), which allows quantitation in less than 30 min. Here, we have tested its use for the analysis of cytokine gene expression in different experimental in vitro and ex vivo settings. First, we compared quantitative competitive RT-PCR with real-time RT-PCR in the quantitation of transcription levels of the CD4(+) cell-specific chemoattractant Interleukin-16 during the maturation of monocyte-derived dendritic cells, and found a good correlation between both methods. Second, differences in the amounts of IL-16 mRNA in synovial tissue from patients with rheumatoid arthritis and osteoarthritis as assessed by real-time RT-PCR paralleled differences in the level of IL-16 protein in the synovial fluid. Finally, we employed real-time RT-PCR to study the cutaneous expression of several cytokines during experimental immunomodulatory therapy of psoriasis by Interleukin-10, and demonstrate that the technique is suitable for pharmacogenomic monitoring. In summary, real-time RT-PCR is a sensitive and rapid tool for quantifying mRNA expression even with small quantities of tissue. The results obtained do not differ from those generated by quantitative competitive RT-PCR.

Topics: Arthritis, Rheumatoid; Cells, Cultured; Dendritic Cells; Gene Expression; Humans; Interferon-gamma; Interleukin-10; Interleukin-16; Interleukin-8; Monocytes; Osteoarthritis; Psoriasis; Receptors, Interleukin-8B; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Synovial Membrane; Tumor Necrosis Factor-alpha

Psoriasis is currently considered to be an inflammatory disease of the skin mediated by T-lymphocytes. The key role in the pathogenesis of psoriasis is played by IL-8 which together with INF-gamma have a mitogenic effect on keratinocytes and trigger their hyperproliferation and activation of neutrofile leucocytes as well as the inflammatory reaction. The skin afflicted with psoriatic lesions contains 100-times increased expression of active IL-8. Its most significant source is probably represented by keratinocytes, but it is produced also by endothelial cells, fibroblasts, T-lymphocytes, polymorphonuclear leucocytes, fibroblasts, Langerhans cells, macrophages. The revealed pathogenetic coincidences render new prospective possibilities in the treatment of psoriasis.

Topics: Humans; Interleukin-8; Keratinocytes; Psoriasis; T-Lymphocytes

The pro-inflammatory properties of interleukin-8 (IL-8) suggest a major role of this peptide in inflammatory processes of skin and other organs. Both biochemical and immunohistochemical studies from our group have demonstrated IL-8 peptide within psoriatic scales and epidermis. So far, however, the relevance of circulating IL-8 and its relation to locally produced IL-8 in this disease remain unclear. Serum IL-8 levels of psoriatic patients were determined in sandwich-ELISA prior to therapy as well as during therapy. Using either the assay from our laboratory or three commercial ELISAs, no correlation was found between serum IL-8 levels and disease severity at any stage of the disease. Similarly, epidermal IL-8-immunoreactivity was monitored immunohistochemically in sequential biopsies from individual psoriatic lesions as they resolved during the course of therapy. Initially, decreased epidermal IL-8 immunoreactivity shifted to a homogeneous staining comparable to normal or non-diseased skin as lesions resolved under treatment. These results indicate a role of IL-8 at local level in psoriatic skin. In contrast to hyperinflammatory diseases like sepsis, where increased serum IL-8 levels are found, in psoriasis either circulating IL-8 is absent or potent mechanisms are operative effectively binding and/or inactivating IL-8 as it enters circulation.

Topics: Biopsy; Humans; Immunohistochemistry; Interleukin-8; Psoriasis; Severity of Illness Index; Skin

Chemokines play an important role in the selective movement of leucocytes into inflammatory areas and they also activate various cells in inflamed tissues. However, it is unclear which cells are the main sources of chemokines in actual inflammatory diseases, even though both mononuclear cells and non-inflammatory resident cells are able to produce chemokines in vitro and the former cells are also the main target of chemokines. To clarify the roles of chemokines that are produced by mononuclear cells in AD, we measured levels in vivo of mRNA for IL-8 and MIP-1 alpha, as well as the level of regulated upon activation normal T cell expressed and secreted (RANTES) mRNA in freshly isolated peripheral blood mononuclear cells from patients with AD. We compared the results with those from psoriatic patients, and patients without AD who were suffering from other cutaneous diseases and eosinophilia. Levels of mRNAs were determined by semiquantitative reverse transcriptase-polymerase chain reactions. Levels of IL-8 and MIP-1 alpha mRNA were elevated not only in atopic patients but also in non-atopic patients with inflammatory skin disease associated with eosinophilia, compared with levels in psoriatic patients and healthy controls. Levels of RANTES mRNA were similar in atopic patients but they were lower in the other two groups of patients when compared with levels in healthy controls. In atopic patients, the levels of both IL-8 and MIP-1 alpha mRNAs but not of RANTES mRNA decreased with improvements in symptom scores after therapy. These findings suggest that mononuclear cells are not only the target of chemokines but might also play an important role in the pathogenesis of AD by producing IL-8 and MIP-1 alpha.

Topics: Adolescent; Adult; Aged; Chemokine CCL4; Chemokine CCL5; Dermatitis, Atopic; Eosinophilia; Female; Humans; Interleukin-8; Leukocytes, Mononuclear; Macrophage Inflammatory Proteins; Male; Middle Aged; Psoriasis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Skin Diseases

Endothelins (ETs), in addition to their systematical activities, exert important functions at the skin level, such as increase of keratinocyte proliferation, neo-angiogenesis and leukocyte chemotaxis, which are among the main characteristics of psoriasis. To assess a possible ET-1 involvement in plaque-type psoriasis, ET-1 determinations were carried out in 15 sera and 8 lesional and non-lesional biopsy skin extracts from psoriatic patients and in 15 sera and 5 biopsy skin extracts from healthy volunteers, sex- and age-matched, using commercially available ELISA kits. A statistical analysis of the results showed that ET-1 levels were increased in sera of psoriatic patients, as compared to normal subjects (p = 0.04). In addition, there was a significant correlation between both serum (r = 0.60, p = 0.02) and lesional skin (r = 0.80, p = 0.03) ET-1 values versus the Psoriasis Area and Severity Index scores. Significant increases of the lesional versus the non-lesional (p = 0.01) and versus the normal (p = 0.04) ET-1 skin extract values were observed, together with a significant correlation between lesional and non-lesional ET-1 skin levels (r = 0.79, p = 0.03). These findings were also confirmed at the mRNA level, using RT-PCR analysis, where increased ET-1 mRNA levels, densitometrically measured, were found in the lesional samples versus non-lesional and normal skin. Since interleukin-8 is involved in psoriasis and shares some biological properties with ET-1, we further evaluated the levels of this cytokine in skin extracts. The behaviour of interleukin-8 paralleled that of ET-1, and a significant correlation between these two molecules was observed in the lesional skin (r = 0.76, p = 0.05). Taken together, these data stress that, as previously described for interleukin-8, ET-1 may be involved in inflammatory processes associated with psoriasis.

Topics: Adult; Aged; DNA Primers; Endothelin-1; Female; Humans; Inflammation; Interleukin-8; Male; Middle Aged; Polymerase Chain Reaction; Psoriasis; Severity of Illness Index; Skin; Transcription, Genetic; Up-Regulation

The chemokine RANTES is a chemoattractant for eosinophils, T lymphocytes of memory phenotype and monocytes, suggesting that it plays an important part in chronic inflammatory and allergic diseases. In various types of cells, RANTES production is markedly induced by tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma in combination. Psoriasis vulgaris is a chronic cutaneous inflammatory disease. Cytokines and chemokines produced by T cells and epidermal keratinocytes, such as interleukin (IL) 8, are involved in the pathogenesis of psoriasis. T-cell clones obtained from psoriatic skin have been shown to produce the Th1 cytokine IFN-gamma. In addition, abnormal expression of proinflammatory cytokines including TNF-alpha has been observed in psoriatic lesions. These reports led us to hypothesis that psoriatic skin could provide epidermal keratinocytes with TNF-alpha and IFN-gamma, so that keratinocytes could produce RANTES. In this study, we addressed the question as to whether RANTES was involved in psoriasis vulgaris. Immunohistochemistry of skin biopsies showed RANTES was present in the intercellular spaces between epidermal keratinocytes, in the fully developed lesions from the middle to the edge of psoriatic plaques, but not in the perilesional uninvolved and healthy control skin. Further, we confirmed the production of RANTES, together with IL-8, by cultured normal human epidermal keratinocytes, using an enzyme-linked immunosorbent assay. Stimulation with TNF-alpha and IFN-gamma in combination synergistically increased the RANTES production in this system. These results clearly demonstrate the expression of RANTES in psoriatic lesions and suggest the involvement of this chemokine in the outcome of cutaneous inflammatory diseases. Tacalcitol (1 alpha,24(R)-dihydroxyvitamin D3), an active vitamin D3 analogue, inhibited RANTES and IL-8 production in cultured normal epidermal keratinocytes. This result indicates that active vitamin D3 is effective in the regulation of chemokine production by epidermal keratinocytes, which may partly account for its action as an antipsoriatic drug.

Topics: Cell Culture Techniques; Chemokine CCL5; Cytokines; Dihydroxycholecalciferols; Dose-Response Relationship, Drug; Humans; Interferon-gamma; Interleukin-8; Keratinocytes; Psoriasis; Recombinant Proteins; Skin; Time Factors; Tumor Necrosis Factor-alpha

Focal infections such as chronic tonsillitis or dental caries occasionally play a role in the induction or exacerbation of palmoplantar pustulosis (PPP). Arthro-osteitis is sometimes a complication in severe cases of PPP. To study the effects of bacterial infection on the exacerbation of cutaneous lesions and arthralgia, we investigated the T-cell receptor V beta repertoire in peripheral blood mononuclear cells (PBMC) and tonsil tissue after tonsillectomy in 4 cases, who had chronic tonsillitis and a history of exacerbation of cutaneous lesions following a sore throat. First, serum levels of interleukin-6 (IL-6) and IL-8 were measured before and after tonsillectomy by enzyme-linked immunosorbent assay (ELISA). Second, 3H-TdR incorporation was used to examine the effects of the culture supernatant on the PBMC of the autologous patients, other PPP patients without tonsillitis and normal controls. T-cell receptor V beta repertoire was examined by the reverse transcriptase-polymerase chain reaction method. Results showed that IL-8 was significantly high in the serum and abundantly released from tonsillar lymphocytes, which may play a role in the accumulation of neutrophils in lesional skin. T-cell receptors V beta 6 and 12 were preferentially expressed on tonsillar lymphocytes, and V beta 4, 7, 9, 17 and 18 were detected relatively frequently. These data suggest that restricted usage of T-cell receptor V beta subsets may play a crucial role in the induction of tonsillitis associated with PPP.

Topics: Adult; Culture Media, Conditioned; Female; Gene Expression; Humans; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Lymphocyte Activation; Male; Middle Aged; Mitogens; Palatine Tonsil; Polymerase Chain Reaction; Psoriasis; Receptors, Antigen, T-Cell, alpha-beta; RNA; Staphylococcus aureus; Superantigens; T-Lymphocytes; Time Factors; Tonsillectomy; Tonsillitis

Interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha) and interleukin-8 (IL-8) production by peripheral monocytes in 15 patients with psoriasis vulgaris and 13 healthy controls were investigated using an enzyme-linked immunosorbent assay. Polymorphonuclear leukocytes, mononuclear leukocytes, monocytes, lymphocytes and serum were isolated and cultured for 1, 12, 24 and 48 h in RPMI 1640 medium without fetal calf serum to release the cytokines. All cytokine levels in serum were very low in both psoriatic patients and the controls. However, a significant increase in the IL-1 alpha levels in the monocyte culture supernatants was observed in the psoriatic patients compared with that of the controls. IL-1 beta and IL-8 levels in the monocyte supernatants were also significantly increased in the psoriatic patients compared with the controls. TNF-alpha levels in the 12-h supernatants of the monocytes increased in some samples from psoriatic patients but not in the 48-h supernatants. Our study confirmed the overproduction of cytokines in the peripheral monocytes of the psoriatic patients and suggest the activation of monocytes in the patients with psoriasis.

Topics: Adult; Aged; Cell Separation; Cell Survival; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-1; Interleukin-8; Lymphocytes; Male; Middle Aged; Monocytes; Neutrophils; Psoriasis; Tumor Necrosis Factor-alpha

Keratinocytes are influenced by cytokines released by skin-infiltrating T lymphocytes. IL-17 is produced by activated CD4+ T cells and can stimulate epithelial cells. We investigated whether IL-17 could modulate the cytokine production and cell-surface molecule expression of keratinocytes. The effects of IL-17 were compared with those of IFN-gamma, which is also derived from activated T cells and is a strong stimulator for keratinocytes. IL-17 enhanced the mRNA and protein production of the proinflammatory cytokines IL-6 and IL-8 in a concentration-dependent way, and induced a weak expression of intercellular adhesion molecule (ICAM)-1 and HLA-DR. The production of IL-1alpha and IL-15 was not altered. IFN-gamma augmented the production of IL-6, IL-8, and IL-15 and strongly induced both cell-surface molecules. IL-17 and IFN-gamma showed marked synergism in the stimulation of IL-6 and IL-8 protein secretion and, to a lesser extent, in the induction of ICAM-1 and HLA-DR expression. The majority of the CD4+ and CD8+ T cell clones derived from lesional psoriatic skin expressed IL-17 mRNA, suggesting that skin-infiltrating T cells can produce this cytokine. This IL-17 mRNA expression was detectable in T helper cell type 1 and type 2 and did not correlate with the IFN-gamma or IL-4 production. In addition, IL-17 mRNA is detectable in biopsies from lesional psoriatic skin, but not in nonlesional control biopsies. Our study indicates that IL-17 is a proinflammatory cytokine, which could amplify the development of cutaneous inflammation and may support the maintenance of chronic dermatoses, through stimulation of keratinocytes to augment their secretion of proinflammatory cytokines.

Topics: CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Clone Cells; Cytokines; Drug Synergism; Female; HLA-DR Antigens; Humans; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-1; Interleukin-15; Interleukin-17; Interleukin-6; Interleukin-8; Keratinocytes; Male; Psoriasis; RNA, Messenger; Skin

The aim of this study is to determine some functions of neutrophil in patients affected by psoriatic arthritis and to compare them to those of patients affected by cutaneous psoriasis and to normal controls. We used a model of experimental cutaneous inflammation allowing to separate a cluster of purified and viable PMN cells. Then we analyzed, within the three groups, the IL-8 concentration in serum and in the supernatant obtained from the inflammatory site to gather data on the possible pathogenic role played by this cytokine in psoriatic arthritis. We studied neutrophil functions in patients with cutaneous psoriasis and psoriatic arthritis, in acute phase, in comparison with healthy control subjects. We investigated in vivo neutrophil migration by Senn's skin window technique and measured adhesion assay and superoxide production in circulating and migrating neutrophils after different stimuli. We also measured IL-8 concentration in serum and in the supernatant obtained from the inflammatory site, artificially created through the skin window scrape. Neutrophil migration in vivo was significantly higher in both groups of patients than in controls. In the presence of fMLP, blood cells showed a burst of superoxide release, which was significantly more pronounced in patients when compared to healthy controls. Neutrophils from skin window scrape showed a much higher response to fMLP as compared to blood cells of all subject groups, but no differences were observed between patients and controls. No correlation was found between the three groups in adhesion ability under basal condition or in response to different stimuli by circulating and migrating neutrophils. Our results also show a great increase of IL-8 in the exudate from patients compared to controls. Our study shows that there is no difference in neutrophil functions between patients with psoriatic arthritis and cutaneous psoriasis; moreover we suggest that the source of high IL-8 levels are neutrophils rather than the keratinocytes.

Topics: Adult; Arthritis, Psoriatic; Case-Control Studies; Cell Adhesion; Cell Movement; Female; Humans; In Vitro Techniques; Interleukin-8; Male; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Psoriasis; Superoxides

The critical role of infiltrating T cells in the pathogenesis of psoriasis is now well established. In order to determine whether circulating T cells from patients with psoriasis were also involved in the disease process, the authors investigated the biological behavior as studied by chemotactic activity of T cells in patients with psoriasis.. A 48 microchemotaxis chamber was employed to determine T-cell chemotaxis activity. In addition, the expression of T cell activation markers such as HLA-DR and interleukin 2 receptor were analysed with fluorescence activated cell sorting technique and serum IL-8 level was measured with ELISA methods. Forty-five patients with psoriasis (23 patients with severe psoriasis and 22 with mild psoriasis) and 21 patients with atopic dermatitis were investigated. For comparison, T-lymphocytes from 20 healthy controls were tested equally.. T-cell chemotactic responses were significantly decreased in patients with severe psoriasis and atopic dermatitis as compared to healthy controls. Increased expression of activation markers such as HLA-DR and interleukin 2 receptor were demonstrated in circulating T cells from psoriatic patients and atopic dermatitis patients in comparison to healthy controls. Serum IL-8 level was significantly increased in patients with psoriasis and atopic dermatitis.. Circulating T cells in patients with severe psoriasis show abnormal in vitro chemotactic response to IL-8. Furthermore, the in vivo activation state of T lymphocytes in these patients and increased level of serum IL-8 seemed to be associated to their decreased in vitro T-cell chemotactic responses.

Topics: Chemotaxis, Leukocyte; Dermatitis, Atopic; Humans; Interleukin-8; Psoriasis; T-Lymphocytes

The biologically active vitamin D analog calcipotriol is effective and safe in the topical treatment of psoriasis, but its exact mechanism of action is unknown.. We investigated expression of 1,25-dihydroxyvitamin D3 receptors, markers for inflammation (CD1a, CD4, CD8, CD11b, CD15; NAP-1/interleukin-8; 55 kd tumor necrosis factor-receptor; intercellular adhesion molecule-1; HLA-DR), proliferation (proliferating cell nuclear antigen, Ki-67), and differentiation (transglutaminase K; involucrin; cytokeratin 16) in psoriatic skin during topical calcipotriol treatment.. For immunohistochemical staining we used the labeled avidin-biotin technique on cryostat-cut sections.. We found a significant increase of 1,25-dihydroxyvitamin D3 receptor expression in epidermal basal keratinocytes of lesional psoriatic skin during calcipotriol treatment. In all patients analyzed, effects on proliferation and differentiation of epidermal keratinocytes were stronger than effects on dermal inflammation. Effects on inflammation were more pronounced in the epidermal than in the dermal compartment.. Our findings indicate that analogs of 1,25-dihydroxyvitamin D3 upregulate their corresponding receptor in human keratinocytes in vivo. This mechanism may be important in the therapeutic efficacy of vitamin D analogs in psoriasis. The differential therapeutic effects in the epidermal and dermal skin compartments may be due to a reduced bioavailability of calcipotriol in the dermal compartment.

Topics: Administration, Cutaneous; Antigens, CD; Antigens, CD1; Biological Availability; Calcitriol; CD11 Antigens; CD4 Antigens; CD8 Antigens; Cell Differentiation; Cell Division; Dermatologic Agents; Epidermis; Gene Expression Regulation; HLA-DR Antigens; Humans; Immunoenzyme Techniques; Immunohistochemistry; Intercellular Adhesion Molecule-1; Interleukin-8; Keratinocytes; Keratins; Lewis X Antigen; Male; Proliferating Cell Nuclear Antigen; Protein Precursors; Psoriasis; Receptors, Calcitriol; Receptors, Interleukin; Receptors, Interleukin-8A; Receptors, Tumor Necrosis Factor; Skin; Transglutaminases; Tumor Necrosis Factor-alpha; Up-Regulation

Topics: Cyclosporine; Humans; Immunosuppressive Agents; Interleukin-1; Interleukin-8; Psoriasis

The potent calciotropic hormone calcitriol (1,25-dihydroxyvitamin D3, 1,25(OH)2D3) has been shown to be very effective and safe in the topical treatment of psoriasis. In vitro, 1,25(OH)2D3 inhibits proliferation and stimulates differentiation of human keratinocytes. Increasing evidence suggests an immunoregulatory function of this potent steroid hormone. To further characterize the biological effects of topical calcitriol treatment in psoriasis, we have analyzed immunohistochemically the expression of markers for epidermal proliferation (proliferating cell nuclear antigen=PCNA) and differentiation (transglutaminase K, involucrin, cytokeratin 16), as well as inflammation (CD1a, 55 kDa TNF-receptor, NAP-1/IL-8) in calcitriol-treated psoriatic skin in situ. Our findings strongly support the hypothesis that calcitriol modulates keratinocyte proliferation/differentiation as well as inflammation in human skin in vivo. The immunoreactivity of markers for epidermal proliferation and differentiation, as well as of CD1a and NAP-1/IL-8, changed after 8 weeks of calcitriol treatment almost completely to the pattern characteristic for non-lesional psoriatic skin, while a large number of 55 kDa TNF-receptor positive cells could be found in the dermal compartment.

Topics: Administration, Topical; Antigens, CD1; Calcitriol; Cell Differentiation; Cell Division; Humans; Immunohistochemistry; Interleukin-8; Keratinocytes; Keratins; Proliferating Cell Nuclear Antigen; Protein Precursors; Psoriasis; Receptors, Tumor Necrosis Factor; Skin; Transglutaminases

Increased levels of several cytokines, mainly proinflammatory mediators, have been reported in psoriatic lesions. Little information, if any, is available concerning other cytokines, especially those initially studied as marrow differentiation agents. Using the experimental approach of measuring cytokines released by skin organ cultures. IL-11 and three other proinflammatory cytokines (IL-1 beta, IL-6 and IL-8) were determined using commercially available ELISA kits in supernatants of ten biopsies from lesional and nonlesional psoriatic skin areas and in supernatants of biopsies from ten normal volunteers. The results obtained showed that the amounts of IL-11 and the other three modulators were significantly increased in the material from the lesional areas (P < 0.01). The amounts of IL-11, which is known to have functional activity similar to the proinflammatory cytokines and to share a receptor component with IL-6, were also correlated with the disease severity index (R = 0.69, P = 0.04). In addition, a nearly significant correlation was noted between the amounts of IL-11 released by the lesional and the nonlesional skin biopsies (R = 0.66, P = 0.05). More detailed studies are needed to clarify whether IL-11 plays a specific functional role in psoriasis, but this study emphasizes the complexity of the pathogenesis of psoriasis and the cytokine network, including activation of proinflammatory and haemopoietic biological response modifiers, in this disease.

Topics: Adult; Aged; Female; Humans; Interleukin-1; Interleukin-11; Interleukin-6; Interleukin-8; Male; Middle Aged; Organ Culture Techniques; Psoriasis; Skin

Topics: Amino Acid Sequence; Biological Assay; Cell Degranulation; Chemokine CCL5; Chemokine CXCL1; Chemokines; Chemokines, CC; Chemokines, CXC; Chemotactic Factors; Chemotaxis, Leukocyte; Chromatography, Gel; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Dermatitis; Electrophoresis, Polyacrylamide Gel; Glucuronidase; Growth Substances; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Leukocytes; Molecular Sequence Data; Peptide Fragments; Psoriasis; Skin

A structural model of interleukin-8 receptor type beta (IL-8R-beta) was constructed based on the structure of bacteriorhodopsin. High temperature molecular dynamics simulations were performed to search the possible conformations of loop regions in IL-8R-beta which recognize the ligand. The crystal structure of interleukin 8 (IL-8) was used as a geometric constraint of the extracellular loop regions of IL-8R-beta in the conformational search. 500 complex structures were extracted from the dynamics trajectory and five plausible models were selected based on the binding energy and known experimental data. To study further the interaction between IL-8R-beta and its ligands, the complex of IL-8R-beta and platelet factor 4 (PF4) C-terminal peptide was also modeled by molecular dynamics simulations. From these models, the N-terminus, extracellular domain 3 and extracellular domain 4 of IL-8R-beta were found to be important for ligand binding. Key residues of these regions involved in ligand binding were characterized. These models provide insight into the structural basis of biological activity of IL-8 and PF4 and may guide the design of potential therapeutic agents targeting IL-8 receptors. Furthermore, the approach developed from this study may have implications for the understanding of other chemokine receptor-ligand interactions that have been recently suggested to be involved in HIV infection.

Topics: Amino Acid Sequence; Anti-HIV Agents; Anti-Inflammatory Agents, Non-Steroidal; Circular Dichroism; Computer Simulation; Crystallography, X-Ray; Drug Design; HIV-1; Interleukin-8; Ligands; Models, Molecular; Molecular Sequence Data; Platelet Factor 4; Protein Binding; Protein Conformation; Psoriasis; Receptors, Chemokine; Receptors, Interleukin; Receptors, Interleukin-8B

The chronic skin disease psoriasis is characterized by epidermal hyperproliferation and inflammation. The exact etiology of the disease is still unknown. At the molecular level, overexpression of growth factors and proinflammatory cytokines such as IL-8 and the corresponding receptor has been described in psoriatic plaques. On the other hand, the loss of inhibitory control mechanisms is involved in the pathogenesis of the disease, as exemplified by the reduced mRNA levels for the cell cycle inhibitor p53 found in lesional skin. Here we extend these findings to a cytokine with negative regulatory functions, IL-10. Only under certain conditions are human keratinocytes able to synthesize IL-10. In skin, pathological overexpression of IL-10 was described om atopic dermatitis. IL-10 exerts its effects via a specific receptor (IL-10R). We show here for the first time the presence and functionality of IL-10R in epidermal cells and its dramatically decreased expression in acute exanthematic psoriatic epidermis by in vitro and in situ binding studies. These results were substantiated using semiquantitative reverse transcriptase-PCR, demonstrating decreased expression of the IL-10R gene in psoriatic skin, its down-modulation by the proinflammatory cytokine IL-8, and its pharmacological induction in cultured cells. Biological responsiveness of epidermal cells toward IL-10 could also be demonstrated by a reduction of the growth rate and inhibition of IFN-gamma-induced HLA-DR expression. Our results provide the first evidence for a role of the IL-10R gene in the homeostasis of the epidermis and substantiate the concept of a loss of negative regulatory peptides as a step in the eruption of psoriasis.

Topics: Acute Disease; Cells, Cultured; DNA; Down-Regulation; Gene Expression Regulation; Glucocorticoids; HLA-DR Antigens; Humans; Interleukin-10; Interleukin-8; Keratinocytes; Polymerase Chain Reaction; Protein Binding; Psoriasis; Receptors, Interleukin; Receptors, Interleukin-10; RNA, Messenger; Up-Regulation

Epidermal infiltration by polymorphonuclear cells is a prominent feature in psoriatic lesions. Expression of neutrophil-specific chemoattractants by lesional keratinocytes could play an important role in the regulation of this infiltration process. We therefore examined the mRNA expression of GRO-alpha, a well-characterized peptide with neutrophil-specific activation profile in psoriatic lesions by in situ hybridization. Clusters of clearly detectable and in some cases highly abundant GRO-alpha hybridization signals could be demonstrated in the differentiated layers of psoriatic epidermis. The signals were clearly associated with keratinocytes, with no nearby neutrophils detectable by microscopic examination. When additional tissue sections of GRO-alpha-expressing lesions were examined with an interleukin-8/neutrophil-activating peptide-8 (IL-8)-specific anti-sense probe, IL-8 expression was detectable and confined to areas also expressing GRO-alpha. Expression of both GRO-alpha and IL-8 is focally upregulated by an as yet unknown mechanism in lesional psoriatic keratinocytes, ultimately leading to neutrophil tissue infiltration. We suggest that the focal expression of GRO-alpha and IL-8 in the epidermal layers above the dermal papillae may be involved in the "squirting papilla" reaction described as a characteristic feature of psoriatic plaque-type lesions.

Topics: Cadherins; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Gene Expression; Growth Substances; Humans; In Situ Hybridization; Intercellular Signaling Peptides and Proteins; Interleukin-8; Keratinocytes; Neutrophils; Psoriasis; RNA Probes; RNA, Messenger; RNA, Neoplasm

Topics: Adenocarcinoma; Aged; Antigens, CD; Base Sequence; DNA Primers; Enzyme-Linked Immunosorbent Assay; Humans; Immunoenzyme Techniques; Interleukin-8; Keratinocytes; Male; Molecular Sequence Data; Monocytes; Polymerase Chain Reaction; Psoriasis; Receptors, Interleukin; Receptors, Interleukin-8A; RNA, Messenger; Sebaceous Gland Neoplasms; Sigmoid Neoplasms; Syndrome

Interleukin(IL-)6 is overproduced in psoriatic lesions. We investigated the contribution of dermal fibroblasts to the local IL-6 production. Fibroblasts (passageno. 2 to 6) derived from lesional psoriatic (PP) and normal human (NN) skin were used to analyse the secretion of IL-6, and the related cytokines IL-1, IL-8 and TNF-alpha, and the expression of their corresponding mRNA by bioassay, ELISA and Northern hybridization, respectively. PP fibroblasts cultured under serum-free conditions produced increased amounts of bioactive IL-6 when compared to NN fibroblasts. Differences were partially restored in the presence of growth factors or serum. The serum-induced IL-6 production reached a maximum within 24 h after seeding and remained unchanged in PP fibroblasts, whereas comparable amounts of IL-6 were produced only 6 days later in NN fibroblasts. There was a clear expression of IL-6 mRNA in both types of fibroblasts under serum-free conditions. Unexpectedly, fetal calf serum, inactivated fetal calf serum as well as human serum completely inhibited the expression of IL-6 mRNA in all the PP fibroblast cultures investigated. NN fibroblasts were clearly less sensitive to this inhibiting effect of serum. Furthermore, medium supplemented with serum-free component or calcium also repressed IL-6 mRNA expression in PP fibroblasts in contrast to NN fibroblasts. Cycloheximide fully restored the repressing effect of serum indicating that serum induced a labile repressor protein. PP and NN fibroblasts produced negligible amounts of IL-1 and TNF-alpha, but the production of IL-8, however, was comparable to that of IL-6. Our results show a differently regulated IL-6 synthesis in PP fibroblasts in vitro, suggesting an active contribution of dermal fibroblasts to the local IL-6 production in psoriasis.

Topics: Blotting, Northern; Cells, Cultured; Culture Media; Culture Media, Conditioned; Cytokines; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Gene Expression; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Kinetics; Psoriasis; Receptors, Cytokine; Reference Values; RNA, Messenger; Skin; Time Factors

Uncontrolled proliferation of epidermal cells is the most prominent characteristic of psoriasis. This widespread skin disease can be effectively treated with the microbial substance FK506, which acts by modulating gene expression. We, therefore, asked if the drug changes the expression of genes involved in growth regulation (the mitogenic cytokine interleukin-8 (IL-8) and p53, a negative cell cycle regulator) and signal transduction (protooncogenes c-ras, c-raf, and HER-2). Gene expression was monitored by semiquantitative mRNA-PCR and for p53 by immunocytochemistry in cultured primary keratinocytes (KC). In addition, p53 expression was analysed in skin biopsies of psoriatic patients. After 1-3 hr, IL-8 mRNA levels were dose-dependently decreased in tacrolimus (FK506)-treated cells. Protooncogene expression was not significantly altered. Interestingly, p53 transcription was clearly induced by FK506 treatment. This tendency could be verified on the protein level by immunocytochemistry. In contrast, p53 expression was decreased in lesional psoriatic as compared to normal skin, providing evidence that not only posttranslational modification of the p53 protein, but also transcriptional modulation of the p53 gene, are involved in pathological processes and pharmacological drug action in skin. Together with earlier results showing downmodulation for IL-8 receptor type A expression in cultured KC treated with FK506, these results suggest that both the mitogenic IL-8/IL-8R system and the cell cycle inhibitor p53 represent potential targets for the antipsoriatic action of the drug, whereas protooncogenes acting downstream in mitogenic signal transduction cascades are unaffected. The differential modulation of an entire set of genes provides evidence for the specificity of the drug effects and rules out nonspecific toxic effects on KC.

Topics: Dose-Response Relationship, Drug; Gene Expression; Genes, Tumor Suppressor; Humans; Immunohistochemistry; Interleukin-8; Polymerase Chain Reaction; Psoriasis; Skin; Tacrolimus

Interleukin-8 (IL-8) may be important in psoriasis as it is expressed in the stratum granulosum, attracts polymorphonuclear cells, and stimulates angiogenesis and keratinocyte mitogenesis. To study intrinsic cutaneous factors in psoriasis, we constructed skin equivalents from psoriatic or adult control fibroblasts with normal foreskin keratinocytes. IL-8 levels were measured in supernatants by enzyme-linked immunosorbent assay and in skin equivalents by immunohistochemistry and in situ hybridization. IL-8 was highly induced in skin equivalents compared to cells grown alone. Epidermal stratification varied among fibroblast lines and was correlated with IL-8 levels, but lesional and nonlesional psoriatic skin equivalents from the same donor were similar. Six fibroblast lines (two psoriasis lesion and four normal) supported only monolayers, while 12 lines (seven psoriasis lesion and five normal) produced stratification. Mean IL-8 levels were significantly lower in dermal equivalents of the first group than the second (0.78 +/- 0.40 vs 3.93 +/- 2.83 ng per ml, mean +/- SD, p = 0.01, analysis of variance). Significantly more IL-8 was secreted by psoriatic than normal fibroblast skin equivalents over 14 d (p = 0.015) with greatest differences at 1 and 4 d. Psoriatic IL-8 levels peaked first and remained increased. IL-8 protein and mRNA were initially strongest in dermal fibroblasts, and at the dermal-epidermal interface. Diffuse epidermal expression was replaced by accentuation in the stratum granulosum. Psoriatic skin equivalents were thicker, had more intense IL-8 staining, and developed invagination. We hypothesize that an IL-8 paracrine loop between fibroblasts and keratinocytes may play a key role in epidermal regeneration in the skin equivalent, in normal wound healing, and in the determination of an intrinsic psoriatic wound-healing phenotype.

Topics: Adult; Aged; Cell Line; Epidermis; Female; Fibroblasts; Humans; Interleukin-8; Keratinocytes; Male; Middle Aged; Psoriasis; RNA, Messenger; Skin; Skin, Artificial; Staining and Labeling

Dense focal accumulation of neutrophils in the upper epidermis is a hallmark of psoriasis. Because the signals for neutrophil diapedesis and migration in vivo are not fully understood, psoriatic lesions with pronounced migration of neutrophils may serve as an important model for studying neutrophil chemotaxis. In this study, we present evidence for differential expression of the neutrophil chemotactic cytokines growth-related oncogene alpha, interleukin-8, and ENA-78 (epithelial cell derived and neutrophil-activating properties, 78 amino acids) in psoriatic lesions. In situ hybridization and immunohistochemistry of serial sections were employed to identify and microanatomically localize the cells producing these chemokines. High levels of focal interleukin-8 message were found to be expressed in the upper epidermis by keratinocytes and, most importantly, neutrophils themselves. Growth-related oncogene alpha transcripts were detected in clusters of keratinocytes of the upper epidermis at the same sites where interleukin-8 mRNA was abundant. In contrast to interleukin-8, growth-related oncogene alpha was also detected in the papillary dermis produced by vessel-associated cells. Sites of interleukin-8 and growth-related oncogene alpha mRNA expression were associated with infiltration of neutrophils. Interestingly, mRNA expression of the highly homologous chemokine ENA-78 was quiescent. In conclusion, our data indicate that growth-related oncogene alpha is an important chemoattractant for neutrophil diapedesis in vivo, whereas further migration of neutrophils and formation of micropustules appears to be influenced by the cooperative action of both growth-related oncogene alpha and interleukin-8.

Topics: Cell Movement; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Growth Substances; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Keratinocytes; Neutrophils; Psoriasis; RNA, Messenger; Skin

The CD40/gp39 pathway is known to be an important feature of B/T cell collaboration leading to T cell-dependent activation, proliferation or differentiation of B cells. Additionally, CD40 is involved in the regulation of B cell survival and apoptosis. Recently, CD40 has been shown to be expressed functionally on non-hematopoietic cells, i.e. endothelial cells. Here, we demonstrate that human keratinocytes (KC) cultured in vitro express CD40 constitutively. The surface expression of CD40 is markedly up-regulated following stimulation with interferon (IFN)-gamma, but not with tumor necrosis factor-alpha or interleukin (IL)-1 beta. This process is regulated at the CD40 mRNA level as demonstrated by Northern blot analysis. Furthermore, ligation of CD40 via soluble gp39, the CD40 ligand, enhances intercellular adhesion molecule (ICAM)-1 and Bcl-x up-regulation on IFN-gamma-stimulated KC, but not lymphocyte function-associated antigen (LFA)-3, B7-2, HLA-DR, or Fas expression. The release of IL-8 is also induced following CD40 ligation on KC. In psoriasis, a T cell-mediated inflammatory skin disease, KC have a markedly enhanced expression of CD40. This expression co-localizes with the expression of ICAM-1, Bcl-x, and an influx of CD3+ T cells. These findings suggest a functional role of CD40 on KC in inflammatory skin disorders such as psoriasis and could make a therapeutic intervention by disrupting the CD40/gp39 pathway an approach to consider in these inflammatory skin diseases.

Topics: Adult; bcl-X Protein; CD3 Complex; CD40 Antigens; Cells, Cultured; Gene Expression; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Keratinocytes; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Psoriasis; RNA, Messenger; Signal Transduction; Skin Diseases

Psoriasis is a common chronic skin disease mediated by cellular immune mechanisms and characterized by an intense neutrophil cell infiltrate and proliferative activation of epidermal keratinocytes. We have previously described the expression of the inducible nitric oxide synthase (iNOS) in epidermal keratinocytes of psoriatic skin lesions. In this study, the role of iNOS in psoriatic inflammation was explored ex vivo in psoriatic skin biopsies and in vitro in primary cultures of human keratinocytes. Messenger RNA for the iNOS enzyme (iNOS mRNA) was detected by reverse transcriptase polymerase chain reaction in skin biopsies from patients with psoriasis, but not in skin specimens from patients with atopic eczema or from healthy volunteers. As demonstrated by in situ hybridization and immunohistochemistry, expression of iNOS mRNA and its gene product was localized to the epidermal keratinocytes of psoriatic skin lesions. In situ hybridization further revealed a complete colocalization of mRNA expression for iNOS with interleukin (IL) 8 receptor-specific mRNA either in the basal germinative cell layer or at focal sites of ongoing neutrophil inflammation in suprabasal cell layers. Because psoriatic keratinocytes have previously been shown to express mRNA transcripts for IL-8, it seemed reasonable to hypothesize that iNOS expression could be induced in an autocrine loop by IL-8. This hypothesis was substantiated by our in vitro experiments showing that a combination of IL-8 and interferon gamma induces the expression of iNOS-specific mRNA and of the functional enzyme in cultured human keratinocytes. These results suggest an important role for iNOS in concert with IL-8 and its receptor early during the formation of psoriatic lesions.

Topics: Biopsy; Cells, Cultured; Dermatitis; Humans; Immunohistochemistry; In Situ Hybridization; Interleukin-8; Keratinocytes; Nitric Oxide Synthase; Polymerase Chain Reaction; Psoriasis; RNA, Messenger

Involvement of T-lymphocytes in the pathogenesis of psoriasis and atopic dermatitis is well established. The question arises as to whether not only tissue infiltrating but also circulating T-lymphocytes are involved in the disease process. Therefore we sought to determine whether T-lymphocytes from patients with psoriasis and atopic dermatitis show abnormal biological behavior to the proinflammatory chemokine interleukin 8 (IL-8) in vitro as studied by their chemotactic activity. In addition, the expression of T-cell activation markers such as HLA-DR and interleukin 2 receptor (IL-2R) were analysed with FACS-technique. In all, 25 patients with psoriasis (13 patients with severe psoriasis and 12 patients with mild psoriasis) and 11 patients with atopic dermatitis were investigated. For comparison. T-lymphocytes from 14 healthy controls were tested equally. The results show that T-cell chemotactic responses to IL-8 were significantly decreased in patients with severe psoriasis as compared to healthy controls. T-cells from patients with atopic dermatitis demonstrated an even more pronounced decrease in chemotactic response as compared to T-cells from psoriasis patients or healthy controls. In contrast, increased expression of activation markers HLA-DR and IL-2R were demonstrated in circulating T-cells from patients with severe psoriasis and atopic dermatitis in comparison to healthy controls. It can be concluded that circulating T-cells in patients with severe psoriasis and atopic dermatitis show a decreased in vitro chemotactic response to IL-8. Furthermore, the in vivo phenotypic activation state of T-lymphocytes in these patients seemed to be associated with their decreased in vitro functional capacity.

Topics: Adolescent; Adult; Aged; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Movement; Chemotaxis, Leukocyte; Dermatitis, Atopic; Female; HLA-DR Antigens; Humans; Interleukin-8; Lymphocyte Activation; Male; Middle Aged; Psoriasis; Receptors, Interleukin-2; Recombinant Proteins; T-Lymphocytes

To evaluate the contribution of peripheral blood monocytes (PBMC) to epidermotropic inflammatory reactions in psoriasis, using an enzyme-linked immunosorbent assay we measured spontaneous interleukin-8 (IL-8) production in PBMC obtained from patients with psoriasis. IL-8 production in the psoriatic PBMC was significantly higher than that in normal control PBMC. Plasma IL-8 levels in psoriatic patients were also moderately increased compared to normal control levels. IL-8 production in PBMC was closely related to the clinical severity of psoriasis and to the response to treatment, including systemic methotrexate (MTX) treatment and tonsillectomy. IL-8 production in PBMC was also positively related to the production of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in these cells. We speculate that the chemotactic cytokine IL-8 contributes to the development of psoriatic skin lesions, and mediates inflammatory reactions from the inflammatory focus to the psoriatic lesions.

Topics: Adolescent; Adult; Aged; Female; Humans; Interleukin-8; Male; Methotrexate; Middle Aged; Monocytes; Palatine Tonsil; Physical Stimulation; Psoriasis; Severity of Illness Index; Time Factors; Tonsillectomy

Keratinocytes from normal and psoriatic skin were tested for their in vitro proliferative response to a range of concentrations of rIL-6, rTGF alpha, rIL-8 and rGM-CSF using a serum-free culture system. With one exception, all normal cultures (11/12) were stimulated by 1000 ng/ml IL-6 (P < 0.001). Six out of ten psoriatic keratinocyte cultures were also stimulated at this concentration, but this just failed to reach significance (P = 0.05). As a group, the response by psoriatic keratinocytes to IL-6 was significantly less than that of normal keratinocytes (P = 0.02). TGF alpha at 1 ng/ml induced proliferation in approximately 60% of both normal (8/12, P < 0.05) and psoriatic (6/10, P < 0.01) keratinocyte cultures; there was no significant difference between the responses of the two groups to this cytokine. In addition, small numbers of both normal and psoriatic cultures responded to TGF alpha over a concentration range of 0.1 to 100 ng/ml. Approximately half of the normal and psoriatic cultures were stimulated by 10-1000 ng/ml IL-8. However, the effect was not significant for the group at any of the concentrations tested. GM-CSF had minimal to no effect on most of the normal and psoriatic cultures tested. This study showed that psoriatic keratinocytes are equally responsive to the stimulatory effects of TGF alpha and IL-8, but are less susceptible to IL-6 compared to keratinocytes from normal skin. These findings are consistent with a role for these cytokines in the maintenance of a hyperproliferative epidermis in psoriasis.

Topics: Adult; Cell Division; Cytokines; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; In Vitro Techniques; Interleukin-6; Interleukin-8; Keratinocytes; Psoriasis; Recombinant Proteins; Transforming Growth Factor alpha

Research into the cause and pathophysiological mechanisms underlying expression of psoriatric skin lesions has been hampered by lack of an appropriate animal model for this common and enigmatic cutaneous disease. These studies characterize normal skin, pre-psoriatic skin, and psoriatic plaque skin samples transplanted onto severe combined immunodeficiency mice. In this report we document that 1), normal, prepsoriatic, and psoriatic plaque keratome skin samples can be transplanted onto severe combined immunodeficiency mice reliably with high rates of graft survival (> 85%) and with reproducible changes consistently observed over prolonged periods of engraftment; 2), after transplantation, by clinical assessment and routine light microscopy, normal skin remained essentially normal whereas pre-psoriatic skin became thicker, and psoriatic plaque skin retained its characteristic plaque-type elevation and scale; 3), by using a panel of antibodies and immunohistochemical analysis, the overall phenotype of human cell types (including immunocytes) that persisted in the transplanted skin was remarkably similar to the immunophenotype of pretransplanted skin samples; 4), clearly recognized interface zones between human and murine skin within the epidermal and dermal compartments could be identified by routine microscopy and immunostaining, with focal areas of chimerism; and 5), elevated interleukin 8 cytokine levels were present in transplanted pre-psoriatic and psoriatic plaque skin samples. We conclude that there are many similarities between pre- and post-transplanted human samples of normal and psoriatic skin that are grafted onto severe combined immunodeficiency mice. Thus, we propose that this new animal model is appropriate for additional mechanistic-type studies designed to reveal the underlying genetic/etiological abnormality, as well as better illuminate the pathophysiological basis, for this important skin disease.

Topics: Animals; Chimera; Disease Models, Animal; Evaluation Studies as Topic; Humans; Immunohistochemistry; Interleukin-8; Mice; Mice, SCID; Psoriasis; Skin; Skin Transplantation; Transplantation, Heterologous

Topics: Antigens, CD1; Autoantigens; Cell Communication; Cytokines; Epidermis; HLA-DR Antigens; Humans; In Vitro Techniques; Interleukin-6; Interleukin-8; Lymphocyte Culture Test, Mixed; Psoriasis; T-Lymphocytes; Tumor Necrosis Factor-alpha

Psoriasis is a common inherited skin disease that is characterized by hyperproliferation of epidermal keratinocytes and excessive dermal angiogenesis. A growing body of evidence supports a key pathogenetic role for activated keratinocytes in the angiogenic response that accompanies psoriasis. We investigated the role of psoriatic epidermis in the aberrant expression of angiogenesis by examining the ability of pure populations of multipassaged keratinocytes obtained from the skin of normal individuals and psoriatic patients to induce angiogenesis in vivo in the rat corneal bioassay and endothelial cell chemotaxis in vitro. Media conditioned by keratinocytes from psoriatic patients, including both symptomless skin and psoriatic plaques, induced vigorous angiogenic responses in over 90% of corneas tested and potently stimulated directional migration of capillary endothelial cells in vitro. In contrast, conditioned medium from normal keratinocyte cultures was weakly positive in less than 10% of corneas assayed and failed to stimulate endothelial cell chemotaxis. Furthermore, keratinocytes from psoriatic skin exhibited a 10- to 20-fold increase in interleukin-8 production and a seven-fold reduction in thrombospondin-1 production. The angiogenic activity present in keratinocyte-conditioned media from psoriatic patients was suppressed by adding either highly purified thrombospondin-1 (125 ng) or following the addition of either normal keratinocyte-conditioned media or neutralizing interleukin-8 antibody. We conclude that psoriatic keratinocytes are phenotypically different from normal keratinocytes with respect to their angiogenic capacity and that this aberrant phenotype is attributable to a defect in the overproduction of interleukin-8 and a deficiency in the production of the angiogenesis inhibitor thrombospondin-1.

Topics: Adult; Angiogenesis Inducing Agents; Animals; Cells, Cultured; Culture Media, Conditioned; Endothelium, Vascular; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-8; Keratinocytes; Male; Membrane Glycoproteins; Middle Aged; Neovascularization, Pathologic; Psoriasis; Rats; Rats, Inbred F344; Skin; Thrombospondins

Topics: Cells, Cultured; Chemotactic Factors; Cytokines; Dermatitis; Humans; Interleukin-1; Interleukin-8; Monocyte Chemoattractant Proteins; Psoriasis; RNA; Skin; Tumor Necrosis Factor-alpha

Interleukin-8 is assumed to play a central role in the pathogenesis of psoriasis. Since an increased expression of the interleukin-8 receptor has been observed both in polymorphonuclear leukocytes and in affected psoriatic epidermis, we were interested in whether the interleukin-8 receptor could be a molecular target of antipsoriatic compounds. Cyclosporine, calcitriol, calcipotriol or dithranol caused a dose-dependent decrease in interleukin-8 binding to cultured human keratinocytes, while interleukin-8 binding to granulocytes was not affected. In addition, the interleukin-8-induced human leukocyte antigen-DR (HLA-DR) expression of keratinocytes was nearly completely blocked by treatment of the cells with these substances. The inhibition of the keratinocyte interleukin-8 receptor and its function by antipsoriatic drugs may contribute to their therapeutic action.

Topics: Anthralin; Calcitriol; Cells, Cultured; Cyclosporine; Flow Cytometry; HLA-DR Antigens; Humans; Interleukin-8; Keratinocytes; Neutrophils; Psoriasis; Receptors, Interleukin; Receptors, Interleukin-8A; Recombinant Proteins

In human fibroblast cultures TPA increased IL-6 and IL-8 production. This was reduced by vitamin D3 metabolites and analogs. The two analogs employed: 1,25 (OH)2-22 (E)-dehydro-24-monohomo vitamin D3 (Compound A) and 1,25 (OH)2 -22 (E)-dehydro-24 dihomo-vitamin D3 (Compound B) may be useful in the therapy of pathologic proliferative disorders including psoriasis, particularly since they are less toxic and have less effect on calcium metabolism than vitamin D3.

Topics: Calcitriol; Calcium; Cell Division; Cell Line; Fibroblasts; Humans; Interleukin-6; Interleukin-8; Psoriasis; Skin

IL-8 is a chemotactic cytokine with proinflammatory and growth-promoting activities. Recently it has been shown to influence several functions of keratinocytes, including HLA-DR expression, chemotaxis, and proliferation by binding to a specific receptor. Because psoriasis vulgaris is characterized by epidermal hyperproliferation and infiltration of inflammatory cells, we investigated the expression of IL-8 and its receptor in normal and psoriatic epidermis using semiquantitative reverse-transcriptase-polymerase chain reaction. In addition the mRNA levels of the proto-oncogenes c-ras, c-raf, c-myc, and HER-2 were also investigated as potential growth-promoting stimuli in psoriatic epidermis. IL-8 mRNA was only detected in lesional psoriatic epidermis, and IL-8R-specific mRNA was found to be 10 times increased in lesional psoriatic epidermis. There was no significant difference in the protooncogene mRNA levels. In order to test the relevance of the massively increased IL-8R levels in psoriatic epidermis, we investigated the effect of the antipsoriatic drug FK-506 on specific IL-8 and IL-8R mRNA expression. FK-506 dose dependently inhibited IL-8R expression and function. Our data suggest that in psoriatic skin, elevated IL-8 levels and markedly increased IL-8R expression may act in concert to induce the cardinal signs of psoriasis--epidermal hyperproliferation and leukocyte infiltration. IL-8R may prove a molecular target for antipsoriatic drugs such as FK-506.

Topics: Adult; Aged; Amino Acid Sequence; Cells, Cultured; Down-Regulation; Female; Gene Expression Regulation; Humans; Interleukin-8; Male; Middle Aged; Molecular Sequence Data; Proto-Oncogenes; Psoriasis; Receptors, Interleukin; Receptors, Interleukin-8A; RNA, Messenger; Skin; Tacrolimus

The purpose of this study was to investigate and to compare, by in situ hybridization, gene expression of IL-1 beta, IL-8, TGF-beta 1, TGF-beta 2, TGF-beta 3, TGF-alpha, p53 and c-myc in lesions and in non-involved skin of patients with psoriasis. All lesional skin biopsies showed overexpression of IL-1 beta, IL-8 TGF-alpha mRNAs. IL-1 beta hybridization signals were strong in a small number of cells localized predominantly in the dermal papillae and in the suprapapillary epidermis. Overexpression of TGF-alpha was observed in all suprabasal keratinocytes, whereas strongly elevated IL-8 mRNA expression was found to be restricted to clusters of suprabasal keratinocytes. TGF-beta 3, p53 and c-myc transcripts were clearly detected in the epidermis of all biopsies, although expression levels were comparable in lesional and non-lesional skin.

Topics: Cytokines; Gene Expression; Genes, myc; Genes, p53; Genes, Tumor Suppressor; Humans; In Situ Hybridization; Interleukin-1; Interleukin-8; Proto-Oncogene Mas; Proto-Oncogenes; Psoriasis; Transforming Growth Factor alpha; Transforming Growth Factor beta

Interleukin (IL)-8 and gro peptides are members of the intercrine-alpha family of chemotaxins known to be present in biologically active form in psoriasis lesions. However, the relative contribution of the three different gro genes to the expression of this material is unknown, as is the stimulus for gro overexpression in psoriatic lesions. To address these questions, Northern blot and semiquantitative polymerase chain reaction analysis were performed on RNA extracted from keratome biopsies of normal skin, untreated plaques of psoriasis, or plaques treated for 1 week with low-dose cyclosporin A (CsA). Northern blot analysis revealed a significant correlation between gro and IL-8 mRNA levels in psoriasis lesions from 26 different individuals (r = 0.61, p = 0.0009), and overexpression of gro was markedly reduced by CsA prior to detectable clinical improvement (79.3%, p = 0.01, n = 22). To determine which form(s) of gro were overexpressed in psoriatic lesions, total keratome RNA (1 microgram) was analyzed by semiquantitative reverse transcription-polymerase chain reaction (SQRT-PCR). In five patients known to markedly overexpress gro and IL-8 mRNAs by Northern blotting, gro-alpha was approximately six times more abundant than gro-beta, and 25 times more abundant than gro-gamma. In cultured human keratinocytes, all three forms of gro mRNA were increased by IL-1 alpha or by interferon (IFN)-gamma plus tumor necrosis factor (TNF)-alpha. However, in contrast to the situation in vivo, CsA had no inhibitory effect on cytokine-stimulated gro expression in cultured keratinocytes. Taken together, these results demonstrate that the gro-alpha gene is selectively overexpressed in psoriatic lesions and strongly suggest that overexpression of gro is a keratinocyte response to activated T cells in psoriasis.

Topics: Base Sequence; Blotting, Northern; Cells, Cultured; Chemokine CXCL1; Chemokines, CXC; Cyclosporine; Gene Expression; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-1; Interleukin-8; Isomerism; Keratinocytes; Molecular Sequence Data; Polymerase Chain Reaction; Proto-Oncogene Proteins; Psoriasis; RNA, Messenger; Skin

Topics: Humans; Interleukin-8; Leukotriene B4; Psoriasis; Skin

Psoriatic scale extracts contain a unique chemotactic peptide fraction that is likely to be involved in the induction of rhythmic transepidermal leukocyte chemotaxis. Recent studies have identified the presence of two unrelated chemotactic peptides in this fraction, ie, C5a/C5a des Arg and interleukin 8 (IL-8), and its related cytokines. To investigate their relative contribution to the transepidermal leukocyte migration as well as their interrelationship in psoriatic lesions, we have quantified concentrations of immunoreactive C5a/C5a des Arg and IL-8 in psoriatic lesional scale extracts and those from related sterile pustular dermatoses such as subcorneal pustular dermatosis and pustulosis palmaris et plantaris.. The concentrations of C5a/C5a des Arg and IL-8 were more significantly increased in the horny-tissue extracts from lesional skin than in those from noninflammatory orthokeratotic skin (P < .01). The increase of C5a/C5a des Arg concentration was specific to the lesional scale extracts, but showed a rather wide range of variation. By contrast, IL-8 concentration, although consistently increased in the lesional scale extracts, was also moderately increased even in noninflammatory scale extracts prepared from ichthyosis vulgaris. The elevation of IL-8 levels in psoriatic lesions was also confirmed by measuring their levels in cutaneous tissue fluid samples collected from suction blisters. However, unexpectedly, some control samples obtained from normal skin also showed a moderate increase in the IL-8 level. Neutrophil chemotactic activity correlated significantly only with the levels of C5a/C5a des Arg in the scales (P < .05). No such significant correlation was found between chemotactic activity and IL-8 or between C5a/C5a des Arg and IL-8.. Based on these results, we speculate that, although IL-8 may exert a synergistic effect with C5a/C5a des Arg in the induction of transepidermal leukocyte chemotaxis, it constitutes a proinflammatory cytokine that is involved in the production of the persistent inflammatory changes characterized by a T-lymphocyte infiltration. In contrast, C5a/C5a des Arg seems to be generated only in the inflammatory lesional skin under specific circumstances that preferentially favor complement activation and also seems to play a major role in the induction of cyclic transepidermal leukocyte chemotaxis from "squirting papillae."

Topics: Adolescent; Adult; Chemotactic Factors; Chemotaxis, Leukocyte; Complement C5a, des-Arginine; Female; Humans; Interleukin-8; Keratosis; Male; Middle Aged; Neutrophils; Psoriasis; Skin

T-cell activation appears to be critical for the maintenance of psoriatic lesions. In this study, we determined whether cytokines released by epidermal cells from psoriatic lesions are providing signals that result in propagation of intralesional T-cell activation. Supernatants were obtained from epidermal cell cultures derived from skin biopsy specimens of psoriatic patients and normal subjects. These supernatants were added to purified normal CD4+ T cells activated via T-cell receptor (immobilized anti-CD3 and fibronectin) or via other activating pathways (anti-CDw60 or UM4D4).. Psoriatic supernatants (n = 9), but not normal supernatants (n = 7, P < .0006), potentiated T-cell stimulation with anti-CD3 and fibronectin to 172% +/- 41% over control stimulation levels. The degree of lesional psoriatic epidermal cell potentiation correlated with the clinical severity of the lesion (r = .82, P = .007). Psoriatic epidermal cytokine potentiation of T-cell activation was not limited to T-cell receptor mediated stimulation; potentiation of anti-CDw60-stimulated CD4+ T cells was also observed. Neutralizing antisera to interleukin 1 and interleukin 8, but not interleukin 6, were found to reduce only partly the observed potentiation of T-cell activation. To determine whether cyclosporine is down modulating T-cell-potentiating cytokine activity in psoriasis, we compared samples obtained during a double-blind clinical trial of intralesional cyclosporine. T-cell-potentiating activity from psoriatic lesional sites treated with cyclosporine was not significantly modulated relative to the activity derived from vehicle-treated or untreated sites.. These data demonstrate that lesional psoriatic epidermal cells release a balance of cytokines that potentiate T-cell activation. Because normal epidermal cells do not potentiate T-cell activation in this system, these findings demonstrate a mechanism by which the epidermis may non-specifically potentiate and perpetuate T-cell activation in psoriatic lesions.

Topics: CD4-Positive T-Lymphocytes; Cell Division; Cells, Cultured; Cyclosporine; Epidermal Cells; Epidermis; Fibronectins; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Interleukins; Keratinocytes; Leukocytes, Mononuclear; Lymphocyte Activation; Psoriasis; Receptors, Antigen, T-Cell; T-Lymphocytes; Up-Regulation

Polymorphonuclear leukocyte (PMNL) infiltration is an important characteristic in psoriatic lesions. The proinflammatory 8-kD peptide interleukin-8 (IL-8) is present in psoriatic scales and possesses a high chemotactic activity on human neutrophils, which may relate to its role in psoriasis. Its chemotactic activity is mediated via specific receptors on PMNL. The goal of our work was to ascertain whether PMNL infiltration in psoriasis can be accounted for by functional abnormalities of the circulating PMNL due to alterations in the IL-8 receptor density or affinity (or both). Results of radioligand binding studies performed in 10 psoriatic patients, 10 patients with atopic eczema and 11 normal controls showed no difference in receptor affinity (Kd) between the groups. However, a slight but significant elevation in IL-8 receptor density was seen on PMNL from psoriatic individuals (31,230 +/- 3,237 binding sites per cell) compared to those from normal volunteers (24,152 +/- 2,643) and atopic eczema patients (24,092 +/- 2,743). Increased number of IL-8 receptors may, besides elevated cutaneous IL-8 concentrations, contribute to the intraepidermal accumulation of PMNL in psoriasis.

Topics: Adult; Aged; Aged, 80 and over; Chemotaxis, Leukocyte; Chronic Disease; Dermatitis, Atopic; Female; Humans; Interleukin-8; Male; Middle Aged; Neutrophils; Psoriasis; Receptors, Immunologic; Receptors, Interleukin-8A

Quantitative studies of cytokine gene expression in vivo are necessary in order to properly describe the cytokine network and to elucidate its role in skin inflammation. Ideally, one should be able to follow cytokine gene expression in epidermal, dermal, and blood compartments. However, such studies are limited by small amounts of available material. Here we report a polymerase chain reaction (PCR) cDNA amplification protocol useful for quantification of specific mRNAs in small skin samples. We found that analysis of dilution series of each sample permitted establishment of quantitative PCR amplification conditions using only picogram to nanogram amounts of total RNA. Cytokine mRNA amounts could then be measured relative to an internal standard species, co-reverse transcribed, and co-amplified with the cytokine species as a measure of cDNA input. Large numbers of samples can be screened rapidly with initial short dilution series identifying cytokine-positive samples and the correct dilution range for each, followed by closer analysis in this range. Epidermal samples obtained through curettage of a small skin area, 2-mm dermal biopsies from the scraped sites, and a few blood drops from the biopsy sites all yielded sufficient RNA for analysis by this protocol. Any mRNA of known sequence can be studied. We analyzed interleukin 8 mRNA levels in more than a hundred epidermal samples from patients and normal test persons and found a variation over several orders of magnitude that seemed to follow the degree of inflammation of the skin.

Topics: Base Sequence; Dermatitis, Contact; DNA; Epidermis; Glyceraldehyde-3-Phosphate Dehydrogenases; Humans; Interleukin-8; Molecular Sequence Data; Polymerase Chain Reaction; Psoriasis; RNA, Messenger; Skin

Topics: Amino Acid Sequence; Chemotactic Factors; Chemotaxis, Leukocyte; Chromatography, High Pressure Liquid; Humans; Interleukin-8; Molecular Sequence Data; Neutrophils; Psoriasis; Sequence Homology, Amino Acid; Skin; Skin Diseases

Previous studies have shown that neutrophil-activating peptide 1/interleukin-8 (IL-8) is present in psoriatic scales and to a lesser extent in normal human epidermis. A panel of monoclonal antibodies and polyclonal antisera raised against IL-8 was used to localize IL-8 with immunoperoxidase techniques in non-lesional and lesional skin of patients with psoriasis and palmo-plantar pustulosis (PPP), and in corresponding sites from healthy subjects. Intracellular IL-8 immunoreactivity was found in all epidermal cell layers in biopsies of healthy subjects and in non-lesional and lesional skin in both PPP and psoriasis. The most intense immunolabeling was regularly found in the basal cell layer. Intercellular epidermal IL-8 immunolabeling was regularly detected in lesional biopsies in PPP and psoriasis, but not in healthy subjects or non-lesional skin in PPP and psoriasis. No intercellular immunolabeling was detected after successful treatment of lesional skin. The majority of cells along the eccrine sweat glands, dermal mononuclear cell infiltrates, and endothelial cells were IL-8 immunoreactive in all biopsies studied. The present study suggests that IL-8, its precursor form, or, alternatively, a degradation product is present in normal human epidermis.

Topics: Humans; Immunohistochemistry; Interleukin-8; Psoriasis; Skin; Sweat Glands

Neutrophil accumulation in the epidermis is a histologic characteristic of psoriasis. We addressed the question: What is the major protein-like chemotactic principle responsible for neutrophil accumulation? Purification of proteinaceous neutrophil chemoattractants from extracts obtained from psoriatic scales by multistep high-performance liquid chromatography (HPLC) yielded three biochemically distinct polypeptides with potent neutrophil chemotactic activity. Aminoterminal amino acid sequence analysis of the quantitatively major neutrophil attractant revealed the sequence ELRXQXIKTYSK, which is identical to that of a 69 residue form of neutrophil-activating peptide-1/interleukin 8 (NAP-1/IL-8). The second major attractant showed the sequence XXVATELRXQXL . . ., which is identical to that of the gene product of the oncogene "gro" as well as "melanoma growth stimulatory activity, MGSA," whereas the third and minor neutrophil chemotaxin has an NH2-terminal sequence identical with NAP-1/IL-8. Estimation of NAP-1/IL-8-related proteins and gro/MGSA by HPLC combined with bioassay revealed a mean of 3.3 +/- 1.7 ng NAP-1/IL-8-related proteins (n = 11) and 3.2 +/- 1.9 ng gro/MGSA (n = 11) per 1 mg psoriatic scales. In normal heel callus (n = 8), these neutrophil attractants were found at concentrations below 0.02 +/- 0.01 ng/mg. The finding of more than 150-times increased amounts of both NAP-1/IL-8 and gro/MGSA in lesional psoriasis material suggest that these mitogenic as well as neutrophil- and lymphocyte-chemotactic compounds may play an important role in the pathogenesis of psoriasis.

Topics: Amino Acid Sequence; Chemotaxis, Leukocyte; Chromatography, High Pressure Liquid; Humans; Interleukin-8; Molecular Sequence Data; Neutrophils; Psoriasis

Interactions between T lymphocytes, neutrophils, and epidermal cells are believed to play a central role in the pathophysiology of psoriasis and other inflammatory cutaneous disorders. Although there is strong evidence that lymphocyte-function-associated antigen-1 (LFA-1) positive T cells are retained in the epidermis via intercellular adhesion molecule-1 (ICAM-1) expression induced on keratinocytes, the molecular basis for the directed migration of T cells or neutrophils towards the epidermis is not known. To investigate whether epidermal keratinocyte-derived products may be important in the migration of T cells and neutrophils into the epidermis, human keratinocytes were cultured in the presence of various cytokines and chemotactic activity of the supernatants were assessed. TNF-alpha stimulation produced directed migrational responses for both neutrophils and T-lymphocytes (both CD4 and CD8), but not B lymphocytes; 69% of T-cell movement and 80% of neutrophil migration induced by the TNF-alpha treated keratinocyte cell supernatants could be inhibited by anti-interleukin-8 (IL-8) serum. Using the same antibody, IL-8 was immunoprecipitated from the supernatants of TNF-stimulated 35S-labelled keratinocytes, and a single 7-kd band product detected by SDS-PAGE. In keeping with these biological activities and protein data, Northern blot analysis of total cellular RNA extracted from keratinocyte monolayers hybridized with a 32P-labelled 1-kb cDNA to IL-8 mRNA, revealed induction of the IL-8 gene in the presence of TNF-alpha and IL-1 beta, but not IFN-gamma. The protein kinase C agonist, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), a known stimulator of psoriasiform cutaneous inflammation when applied directly to murine epidermis, strongly induced keratinocyte elaboration of IL-8 mRNA. These studies demonstrate that activated human keratinocytes are capable of producing biologically active IL-8, and provide evidence that keratinocytes can play a key role in mediating the influx of T cells and neutrophils into the epidermis.

Topics: Blotting, Northern; Cell Adhesion Molecules; Cells, Cultured; Chemotaxis; Cytokines; DNA; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Gene Expression; Humans; Immunohistochemistry; Intercellular Adhesion Molecule-1; Interleukin-8; Keratinocytes; Lymphocyte Function-Associated Antigen-1; Neutrophils; Precipitin Tests; Psoriasis; RNA, Messenger; T-Lymphocytes; Tetradecanoylphorbol Acetate

Various cytokines have in the past been detected in human skin. Among these, the neutrophil-activating peptide NAP-1/IL-8 is a potent 8-kD proinflammatory peptide that has been purified from psoriatic scales. Its chemotactic activity on human neutrophils, as well as its presence in psoriatic scales, may relate to a role in this disease. In the present study, the tissue distribution of the peptide was examined immunohistochemically using two monoclonal antibodies (52E8, 46E5) recently produced and characterized in our laboratory. Immunoreactivity was detected in both normal and psoriatic skin, resulting in uniform suprabasal keratinocyte staining in normal skin with 52E8 and of all keratinocytes with 46E5. Immunoreactivity in psoriasis correlated to the inflammatory tissue reaction, varying from uniform absence in highly active psoriasis to focally weak staining in plaque type psoriasis. Cells of the acrosyringium and hair follicles were always positive and were unaffected by the inflammatory activity. Epidermal immunoreactivity detected in this study may be associated with closely related peptides of the IL8 family or with truncated or extended forms of NAP-1/IL-8.

Topics: Antibodies, Monoclonal; Humans; Immunoenzyme Techniques; Interleukin-8; Psoriasis; Reference Values; Skin; Staining and Labeling

The importance of immunologic mechanisms in psoriasis has been deduced from the ability of immunosuppressive therapies to ameliorate this common and chronic skin disease. Certainly the histology of psoriatic lesions suggests a dialogue between the hyperplastic keratinocytes and infiltrating T lymphocytes and macrophages. To begin dissecting the cytokine network involved in the pathophysiology of psoriasis, the location, in both epidermal and dermal compartments, of tumor necrosis factor-alpha, interleukin-8, intercellular adhesion molecule-1, and transforming growth factor-alpha at the protein and/or mRNA levels were identified. Tumor necrosis factor-alpha was selected as a potentially key regulatory cytokine, first because it induces cultured keratinocyte interleukin-8, intercellular adhesion molecule-1, and transforming growth factor-alpha production, and second because intercellular adhesion molecule-1 expression by keratinocytes in psoriatic epidermis had been identified previously. Using immunohistochemical localization, tumor necrosis factor-alpha was identified in 12 psoriatic lesions as intense and diffuse expression by dermal dendrocytes (macrophages) in the papillary dermis (without significant staining of endothelial cells, mast cells, or dermal Langerhans cells), and focally by keratinocytes and intraepidermal Langerhans cells. Functional interaction between the dermal dendrocytes and keratinocytes was suggested by the presence of interleukin-8 expression of suprabasal keratinocytes immediately above the tumor necrosis factor-alpha-positive dermal dendrocytes. Interleukin-8 mRNA and transforming growth factor-alpha mRNA were detectable in the epidermal roof of psoriatic lesions, but neither was detectable at the protein or mRNA levels in any normal skin specimens. Treatment of cultured human keratinocytes with phorbol ester (which experimentally produces psoriasiform changes on mouse skin) or tumor necrosis factor-alpha also increased interleukin-8 and transforming growth factor-alpha mRNAs. Further elucidation of the cellular and molecular basis for the genesis and evolution of psoriasis will provide the framework for a better evaluation of the cause and treatment of this skin disease.

Topics: Adult; Cells, Cultured; Female; Humans; Immunohistochemistry; Interleukin-8; Keratinocytes; Male; Middle Aged; Psoriasis; Reference Values; RNA, Messenger; Skin; Staining and Labeling; Tissue Distribution; Tumor Necrosis Factor-alpha

In order to better understand the factors regulating disease promotion and activity in psoriasis (PS), we searched for the in situ expression of mRNA for various cytokines in long-standing PS skin lesions. Specific hybridization with a NAP-1/IL-8 anti-sense RNA probe was keratinocyte associated and yielded strong and specific signals exclusively in the upper layers of the lesional epidermis, but not in uninvolved skin from psoriatic patients or normal skin from non-psoriatics. Interestingly, NAP-1/IL-8 transcripts were focally clustered in a spotty pattern predominantly between the tips of elongated papillae, but were absent in the lower epidermal region and the dermal compartment. We consistently failed to detect appreciable numbers of TNF-alpha and/or IL-6 mRNA-containing cells in psoriatic lesions. These results support the notion that IL-8, rather than IL-6, is an important disease-promoting cytokine in PS. In view of the known in vitro and in vivo effects of IL-8, it is conceivable that this substance greatly contributes to the major pathologic changes seen in psoriatic skin, i.e., keratinocyte hyperproliferation and leucocyte infiltration. In this case, local pharmacologic down-regulation of NAP-1/IL-8 activity could be a promising therapeutic strategy in PS.

Topics: Cell Adhesion Molecules; E-Selectin; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Keratinocytes; Nucleic Acid Hybridization; Psoriasis; RNA, Messenger

Neutrophil-activating peptide 1/interleukin 8 (NAP-1/IL-8) is a recently described cytokine with potent chemotactic activity for human neutrophil granulocytes (PMN) and T cells. In psoriasis, a chronic hyperproliferative and inflammatory skin disorder, PMN and T cells are found as prominent cells in the inflammatory infiltrate of the lesions; however, monocytes were shown to be the first cells invading a newly formed plaque. NAP-1/IL-8 was found to be present in high amounts in the skin and in scale material of psoriatic patients. Psoriasis responds well to systemic treatment with cyclosporin A (CsA), an immunosuppressive peptide. Therefore, we addressed the question of whether the clinical improvement of psoriatic patients during CsA therapy may be due to an inhibition of NAP-1/IL-8 production and secretion from monocytes. Purified human monocytes were stimulated by lipopolysaccharide in the presence or absence of various concentrations of CsA. Production of NAP-1/IL-8 was determined as expression of specific mRNA by fluorescent in situ hybridization. Secreted peptide was measured by bioassay (PMN chemotaxis) and enzyme-linked immunosorbent assay (ELISA) using specific monoclonal antibodies. The results show that CsA neither inhibited mRNA expression for NAP-1/IL-8 nor secretion of the peptide. These findings support the hypothesis that the pharmacological effect of CsA may be restricted to the inhibition of T-cell activation and proliferation.

Topics: Cyclosporins; Gene Expression; Humans; In Vitro Techniques; Interleukin-8; Monocytes; Psoriasis; RNA, Messenger

Topics: Humans; Interleukin-8; Psoriasis; Skin

Migration of polymorphonuclear leukocytes (PMN) into the upper layers of involved epidermis represents a characteristic feature of psoriasis. By analysis of psoriatic scale material we were able to identify two potent proinflammatory peptides, which are present in the upper epidermis from psoriatic lesions. Both factors (C5ades arg and NAP) show strong chemotactic activity for human neutrophils in vitro as well as in vivo. Whereas C5ades arg is a known mediator activated by either alternative or classical activation of the complement cascade, NAP represents a newly detected peptide with a molecular weight of 8,000 daltons which is produced by a variety of cells participating in the psoriatic tissue reaction.

Topics: Cell Movement; Chemotactic Factors; Chromatography, Ion Exchange; Complement C5; Complement C5a, des-Arginine; Epidermis; Inflammation; Interleukin-8; Neutrophils; Peptides; Psoriasis

Topics: Chemotactic Factors; Chemotaxis, Leukocyte; Humans; Interleukin-8; Leukotriene B4; Molecular Weight; Psoriasis

1. Lipid extracts of scale from the lesions of the skin disease psoriasis were purified by high performance liquid chromatography (h.p.l.c.). Assay of fractions by an agarose microdroplet method showed the presence of a novel neutrophil chemokinetic compound which possessed the chromatographic properties of a monohydroxy fatty acid, yet was distinct from the chemoattractant eicosanoid, 12-hydroxyeicosatetraenoic acid, previously isolated in psoriasis. 2. The novel, material, termed compound X, was also detected in fractions collected on h.p.l.c. of extracts of chamber fluid samples obtained from abraded psoriatic lesions, but was not detectable in samples from clinically normal skin. 3. Comparison of the straight and reversed phase h.p.l.c. retention times of compound X with those of a range of standard monohydroxy fatty acids, together with further analysis by gas chromatography-mass spectrometry and assay of selected standards for neutrophil chemokinetic activity, failed to reveal the structural identity of compound X. 4. The finding of a further compound in psoriatic lesions, which stimulates neutrophil movement, highlights the complexity of inflammatory mediator production in this disease.

Topics: Adult; Chemotactic Factors; Chromatography, High Pressure Liquid; Eicosanoic Acids; Female; Gas Chromatography-Mass Spectrometry; Humans; Interleukin-8; Male; Neutrophils; Psoriasis

The IL-1-like neutrophil chemoattractant activity previously reported by us to be present in the stratum corneum of psoriatic skin lesions has now been characterized further. Aqueous extracts of stratum corneum samples from psoriatic lesions and from the heels of normal volunteers were ultrafiltered to yield 10- to 30-kDa fractions. The ultrafiltered psoriatic preparations consistently contained greater neutrophil chemokinetic activity than the normal heel preparations, but in contrast the latter contained markedly greater IL-1 activity than the former. Successive chromatographic purification of psoriatic lesional stratum corneum extracts showed that the neutrophil chemokinetic material previously reported to co-elute with IL-1 activity on reversed phase HPLC, but to be distinct from C5a des arg, could now be separated by anion exchange HPLC into at least four different chemokinetic compounds that were also resolved from the IL-1 activity. The reversed phase HPLC-purified chemokinetic material from psoriatic stratum corneum was also active in a neutrophil chemotaxis assay. These findings show that samples from psoriatic skin lesions contain a group of novel 10- to 30-kDa neutrophil chemoattractant compounds that are distinct from both C5a des arg and IL-1. The contrasting neutrophil chemokinetic and IL-1 activities in psoriatic lesional and normal heel stratum corneum preparations support the finding that the two activities are produced by different compounds. These neutrophil chemoattractant and IL-1-like compounds may be of pathogenic importance in inflammatory skin disease.

Topics: Adolescent; Adult; Aged; Cell Line; Chemotactic Factors; Chemotaxis; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Female; Humans; Interleukin-1; Interleukin-8; Male; Middle Aged; Molecular Weight; Neutrophils; Psoriasis; Skin; Thymidine; Tissue Extracts; Ultrafiltration

Increased amounts of 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) are found in the lesional skin of patients with the skin disease psoriasis when compared to clinically normal skin. Stereochemical analysis has recently shown that the 12-HETE present in lesional psoriatic scale is the (R), and not the (S) hydroxyl enantiomer, produced by platelets. Since the chemoattractant activity of 12(R)-HETE has not previously been described, the (R) and (S) hydroxyl enantiomers of 12-HETE have now been synthesised and their chemokinetic activity compared in vitro. 12(R)-HETE, was more potent than 12(S)-HETE as a chemokinetic agent for human polymorphonuclear leucocytes but 2000 times less potent than leukotriene B4. In contrast to results obtained with the 12-HETE enantiomers, the chemoattractant compound 5(S)-HETE was found to be more potent than the 5(R) hydroxyl enantiomer. Thus, the configuration of the hydroxyl group appears to be of importance to the chemokinetic activity of the HETEs, and the increased potency of the 12(R) enantiomer may enhance its significance as a mediator of inflammation in psoriasis.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Chemotactic Factors; Humans; Hydroxyeicosatetraenoic Acids; In Vitro Techniques; Interleukin-8; Kinetics; Neutrophils; Psoriasis; Stereoisomerism

Scales from patients with nonpustular psoriasis were investigated for the presence of peptides capable of activating functional activities in human polymorphonuclear leukocytes (PMNL). Two compounds with similar molecular weight (12,500 and 15,000) were isolated which markedly stimulated PMNL functional activities including chemotaxis, generation of superoxide radical anion (O-2), and liberation of beta-glucuronidase as a marker enzyme. As revealed by ion-exchange and subsequent radioimmunoassay followed by chromatofocusing, one peptide proved to be the desarginated form of the complement split product C5ades arg. No C5a was detectable. As a second psoriatic scale chemotaxin we isolated an anionic neutrophil-activating peptide (ANAP) which shows a single isoelectric point at pH 6.8. This peptide shares some of the characteristics of epidermal cell-derived thymocyte-activating factor and interleukin 1 and, as shown by deactivation experiments, it cross-reacts with a monocyte-derived cytokine. The 2 newly described neutrophil-activating peptides (C5ades arg and ANAP) may play an important role in the psoriatic tissue reaction.

Topics: Cell Movement; Chemotactic Factors; Chemotaxis, Leukocyte; Chromatography, Gel; Chromatography, Ion Exchange; Complement C5; Complement C5a, des-Arginine; Glucuronidase; Humans; Interleukin-8; Isoelectric Focusing; Lactoferrin; Molecular Weight; Neutrophils; Peptides; Peroxidase; Psoriasis; Superoxides