interleukin-8 and Dermatitis--Atopic

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 cutaneous lymphocyte-associated antigen (CLA) is a carbohydrate epitope present on memory/effector T cells that infiltrate inflamed skin. E-selectin is the ligand for CLA and is induced under inflammation on endothelial cells. CLA was originally postulated as a phenotype marker for skin-associated T cells. We studied the specific in vitro response to skin-associated allergens of CLA+ and CLA-CD45RO+ T cells in atopic dermatitis (AD) and contact dermatitis (CD), which represent two well-characterized T cell-mediated cutaneous allergic inflammations. Whereas CLA+ T cells from AD patients preferentially responded to house dust mite (HDM) and CLA+ T cells from nickel CD patients showed an increased response to nickel, CLA-T cells showed very little response in both cases. In contrast, tetanus toxoid, a systemically acting antigen, induced a proliferative response in both CLA+ and CLA- cells. Interestingly the response to HDM in patients with asthma +/- AD was preferentially found in the CLA- subset indicating the involvement of different homing receptors for mucosal tissues. Moreover, CLA+ T cells showed enhanced migration through activated human umbilical vein endothelial cell monolayers compared to CLA- T cells. The CLA binding to E-selectin is initially responsible for the extravasation that also involves VLA-4/VCAM-1 and LFA-1/ICAM-1 interactions. We have recently identified IL-8 as an endothelial cell-derived chemokine and the IL-8 receptor type B which control CLA+ T cell migration. Such a CLA-mediated migration would localize memory/effector T cells that respond to antigens and reach the body through inflamed skin. Our data support the existence of a regionalization of the immune system and in particular of the skin immune system. It may allow an efficient distribution of the immune defense to different sites of the body.

Topics: Animals; Antigens, Differentiation, T-Lymphocyte; Antigens, Neoplasm; CD3 Complex; Dermatitis, Atopic; Dermatitis, Contact; E-Selectin; Epitopes; Humans; Immunologic Memory; Intercellular Adhesion Molecule-1; Interleukin-8; Leukocyte Common Antigens; Lymphocyte Function-Associated Antigen-1; Membrane Glycoproteins; Mites; Nickel; T-Lymphocytes; Vascular Cell Adhesion Molecule-1

For effective management of atopic dermatitis (AD), it is important to introduce a therapeutic agent, which although having the fewest side effects, has the greatest anti-inflammatory effect. In the course of screening anti-inflammatory agents, we obtained BSASM, a mixture of several plant extracts. This study was designed to investigate the AD-mitigating effect of BSASM in patients, as well as its anti-inflammatory and immunomodulatory effects in an in vitro experiment. The anti-inflammatory effects of BSASM were evaluated by the level of production of proinflammatory cytokines. Clinical evaluation was also done using eczema area severity index (EASI) score in AD patients. BSASM inhibited LPS-induced activation of NF-kappaB promoter. In addition, LPS-induced an increase of IL-8, and the TNF-alpha production in THP-1 cells was also inhibited. These results suggest that BSASM has an anti-inflammatory activity. A clinical study in patients with AD showed that BSASM induced a reduction of EASI score, degree of pruritus, and TEWL on both the antecubital fossa and abdomen. Besides, BSASM had no irritative or allergic effects. Based on these results, we conclude that BSASM has anti-inflammatory and AD-mitigating effects.

Topics: Adjuvants, Immunologic; Adolescent; Anti-Inflammatory Agents; Cell Line; Child; Cytokines; Dermatitis, Atopic; Drugs, Chinese Herbal; Female; Humans; Interleukin-2; Interleukin-8; Lipopolysaccharides; Male; NF-kappa B; Phytotherapy; Plant Extracts; Time Factors; Tumor Necrosis Factor-alpha

In order to further evaluate the role of cytokines in the induction of atopic pruritus, leukocytes from 10 atopic eczema patients or 10 nonallergic controls were stimulated in vitro with mite or birch pollen antigen for 1 and 4 days. Subjects were prick-tested with the supernatants, and whealing and itching were evaluated 20 and 60 min later. The supernatants were also examined for the contents of GM-CSF, IL-2, IL-6 and IL-8 by ELISA and TNFalpha. Two hours prior to testing, the antihistamine cetirizine (20 mg) or a placebo tablet were given to the patients according to a randomized, double-blind study protocol. After pricking with antigen-stimulated leukocyte supernatants, 6 of 10 patients but no controls reacted mostly at 20 min with whealing and/or pruritus. In the cetirizine-treated group, no decrease in these skin reactions was seen compared to placebo. Analysis for cytokines showed increased levels of IL-8 in allergen-stimulated samples, with no correlation to the induction of itching or whealing by these supernatants. IL-6 levels were low and variable, and GM-CSF, IL-2 and TNFalpha levels were always below standard values. These data show that leukocytes selectively release IL-8 in response to in vitro antigen stimulation. They furthermore provide additional support for the concept that as yet to be identified products play a role in atopic pruritus.

Topics: Adolescent; Adult; Allergens; Animals; Anti-Allergic Agents; Antigens; Cetirizine; Cross-Over Studies; Culture Media, Conditioned; Cytokines; Dermatitis, Atopic; Dose-Response Relationship, Drug; Double-Blind Method; Enzyme-Linked Immunosorbent Assay; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hypersensitivity, Immediate; Interleukin-2; Interleukin-8; Male; Middle Aged; Mites; Pollen; Pruritus; Severity of Illness Index; Skin Diseases; Skin Tests; Time Factors; Tumor Necrosis Factor-alpha

Atopic dermatitis is a T-cell mediated inflammatory skin disease with detected elevated levels of histamine in skin or plasma. In this study, the effects of histamine in a T

Topics: alpha-Tocopherol; Cytokines; Dermatitis, Atopic; Histamine; Humans; Interleukin-8; Keratinocytes; RNA, Messenger; Skin; Tocopherols

Atopic dermatitis (AD) is a skin disease characterized by pruritus. The present study aimed to discover a herbal combination with anti-allergic and anti-inflammatory activities to treat AD. First, the anti-allergic and anti-inflammatory activities of herbs were evaluated by RBL-2H3 degranulation and HaCaT inflammatory models. Subsequently, the optimal proportion of herbs was determined by uniform design-response surface methodology. The effectiveness and synergistic mechanism was further verified. Cnidium monnieri (CM) suppressed β-hexosaminidase (β-HEX) release, saposhnikoviae radix (SR), astragali radix (AR), and CM inhibited the release of IL-8 and MCP-1. The optimal proportion of herbs was SR∶AR∶CM = 1: 2: 1. The in vivo experiments results indicated that the topical application of combination at high (2 ×) and low (1 ×) doses improved dermatitis score and epidermal thickness, and attenuated mast cell infiltration. Network pharmacology and molecular biology further clarified that the combination resisted AD by regulating the MAPK, JAK signaling pathways, and the downstream cytokines such as IL-6, IL-1β, IL-8, IL-10, and MCP-1. Overall, the herbal combination could inhibit inflammation and allergy, improving AD-like symptoms. The present study discovers a promising herbal combination, worthy of further development as a therapeutic drug for AD.

Topics: Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Cnidium; Cytokines; Dermatitis, Atopic; Humans; Interleukin-8; Mice; Mice, Inbred BALB C; Skin

The cytokine profile of atopic dermatitis (AD) depends on age, ethnicity, and disease severity. This study examined biomarkers in children with AD collected by tape strips and skin biopsies, and examined whether the levels differed with filaggrin genotype, disease severity, and food allergy.. Twenty-five children aged 2-14 years with AD were clinically examined. Skin biopsies were collected from lesional skin and tape strips were collected from lesional and non-lesional skin. We analyzed natural moisturizing factor (NMF) and 17 immune markers represented by mRNA levels in skin biopsies and protein levels in tape strips. Common filaggrin gene mutations were examined in all children.. The cytokine profile in lesional skin was dominated by a T helper (Th) 2 response in skin biopsies, and by a general increase in innate inflammation markers (interleukin (IL)-1α, IL-1β, IL-8, IL-18) along with TARC and CTACK in tape strips. The levels of TARC, CTACK, IL-8, IL-18 showed significant correlation with AD severity in both lesional and non-lesional tape stripped skin, while no significant correlations were observed in skin biopsy data. In tape strips from lesional and non-lesional skin, the levels of NMF and selected cytokines differed significantly between children with and without FLG mutations and food allergy.. Sampling of the stratum corneum with non-invasive tape strips can be used to identify biomarkers that are associated with disease severity, food allergy and FLG mutations. Skin biopsies showed robust Th2 signature but was inferior for association analysis regarding severity.

Topics: Biomarkers; Biopsy; Child; Cytokines; Dermatitis, Atopic; Filaggrin Proteins; Humans; Interleukin-18; Interleukin-8; Skin

No biomarkers have been identified that can classify subtypes of hand eczema (HE). Although skin biopsies represent the gold standard for investigations of the skin, the invasive technique is not favorable when investigating skin from sensitive areas. Recent advances in the use of skin-tape strips for molecular investigations enable noninvasive investigations of HE.. By using whole transcriptome sequencing (WTS), the molecular profile of HE according to different localizations on the hands, etiologies, and clinical/morphological subtypes was investigated.. Thirty adult, Danish HE patients, 12 with and 18 without concurrent atopic dermatitis (AD), as well as 16 controls were included. Tape strip samples were collected from lesional, nonlesional, and healthy skin. Total RNA was extracted and WTS was performed.. The largest molecular difference of HE patients with and without AD was found in nonlesional skin areas and included a downregulation of CXCL8 for HE patients without AD. Differences between allergic and irritant contact dermatitis included epidermal biomarkers such as EPHA1.. Skin tape strip samples could be used to assess the gene expression profile of HE on different localizations of the hands. The skin tape strip method identified new molecular markers that showed promising result for the identification of HE subtypes.

Topics: Adult; Aged; Biomarkers; Dermatitis, Allergic Contact; Dermatitis, Atopic; Dermatitis, Irritant; Diagnosis, Differential; Down-Regulation; Exome Sequencing; Female; Hand Dermatoses; Humans; Interleukin-8; Male; Middle Aged; Receptor, EphA1; Skin; Specimen Handling; Surgical Tape; Transcriptome

The stratum corneum contains several growth factors and cytokines that are synthesized in keratinocytes. We previously reported that the amount of interleukin-8 in the stratum corneum (scIL-8) is related to the severity of local skin inflammation in atopic dermatitis (AD). However, it is unknown whether scIL-8 levels reflect pharmacologic responses to a therapeutic intervention in AD patients. Therefore, in this study, we aimed to investigate whether the improvement of dermatitis in AD is correlated with scIL-8 levels before and after topical corticosteroid treatment.. Stratum corneum samples were collected from 22 AD patients using the noninvasive tape-stripping method before treatment, 2 weeks after topical treatment, and 4-6 weeks after treatment.. scIL-8 levels on the forearm reduced significantly from 790 ± 348 pg/mg before treatment to 163 ± 68 pg/mg 2 weeks after treatment and 100 ± 37 pg/mg 4-6 weeks after corticosteroid treatment. scIL-8 levels on the abdomen also reduced significantly from 902 ± 391 to 142 ± 38 pg/mg at the end of study. The reduction in scIL-8 levels was associated with the improvement in local skin severity in AD. We also found that scIL-8 levels, along with blood biomarker levels (serum thymus and activation-regulated chemokine, lactate dehydrogenase, and %eosinophil), decreased significantly after the treatment.. The scIL-8 concentration decreases with improvements in skin symptoms in AD patients after topical corticosteroid treatment; thus, it may be a suitable biomarker for monitoring therapeutic effects in AD patients.

Topics: Adolescent; Adult; Biomarkers; Combined Modality Therapy; Dermatitis, Atopic; Disease Management; Disease Susceptibility; Female; Humans; Interleukin-8; Male; Prognosis; Severity of Illness Index; Skin; Treatment Outcome; Young Adult

Topics: Animals; Anti-Inflammatory Agents; Artemisia; Cell Survival; Chemokines; Dermatitis, Atopic; Humans; Inflammation; Interferon-gamma; Interleukin-6; Interleukin-8; Keratinocytes; Male; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; Plant Extracts; Skin; STAT1 Transcription Factor; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Chronic skin inflammation in atopic eczema is associated with elevated expression of proinflammatory genes and activation of innate immune responses in keratinocytes. MicroRNAs (miRNAs) are short, single-stranded RNA molecules that silence genes via the degradation of target mRNAs or inhibition of translation. Recent studies have demonstrated that miR-124 is associated with regulation of inflammation factors in several diseases. The aim of this study was to investigate the role of miR-124 in skin inflammation of atopic eczema. We showed that miR-124 expression is decreased in chronic lesional skin of patients with atopic eczema, and could be strongly inhibited by IFN-γ and TNF-α. Through Western blot, real-time PCR and luciferase assays, we revealed that miR-124 inhibited the expression of p65, a member of NF-κB family which can regulate many factors involved in the immune response and inflammatory reactions, through direct targeting. Further, upon IFN-γ or TNF-α stimulation, IL8, CCL5 and CCL8 showed to be significantly upregulated by IFN-γ or TNF-α, downregulated by miR-124; the promotive effect of IFN-γ and TNF-α could be partially reversed by miR-124. The levels of IL8, CCL5 and CCL8 could be significantly downregulated by p65 knockdown, upregulated by miR-124 inhibition; the suppressive effect of p65 knockdown could be partially reversed by miR-124. Moreover, contrary to miR-124, p65, IL8, CCL5 and CCL8 mRNA expression was upregulated in chronic lesional skin of patients with atopic eczema, and all inversely correlated with miR-124. Taken together, our data demonstrate that miR-124 controls NF-κB-dependent inflammatory responses in keratinocytes and chronic skin inflammation in atopic eczema; rescuing miR-124 expression presents a promising strategy for atopic eczema treatment.

Topics: Base Sequence; Chemokine CCL5; Chemokine CCL8; Dermatitis, Atopic; Gene Expression Profiling; Gene Expression Regulation; Humans; Interferon-gamma; Interleukin-8; Keratinocytes; MicroRNAs; Primary Cell Culture; RNA, Small Interfering; Signal Transduction; Skin; Transcription Factor RelA; Tumor Necrosis Factor-alpha

Vascular endothelial growth factor (VEGF) was found increased in the stratum corneum of patients with atopic dermatitis (AD). However, its potential pathogenic role(s) in AD needs further clarification.. The aim of this study was to determine whether VEGF serum levels correlate with other selected cytokine levels and features of AD.. VEGF and other cytokine levels were measured in 83 patients with AD and in a control group and then correlated with clinical and laboratory parameters of AD.. The mean serum concentrations of VEGF and tumor necrosis factor α were significantly higher in patients with AD than in the control group, whereas the mean interleukin eight serum level was lower. VEGF concentrations correlated with the severity of AD as expressed by SCORAD index and objective SCORAD.. VEGF could be regarded as a potentially important mediator in the pathogenesis of AD, as VEGF levels correlate somewhat with AD severity.

Topics: Adolescent; Adult; Case-Control Studies; Chemokine CCL5; Cytokines; Dermatitis, Atopic; Female; Humans; Immunoglobulin E; Interleukin-15; Interleukin-6; Interleukin-8; Male; Middle Aged; Severity of Illness Index; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Young Adult

IL-9 and its receptor play important roles in the pathogenesis of asthma. Its role in atopic dermatitis (AD) was examined in just a few studies, including nucleotide polymorphisms, increased transcriptional levels of IL-9 and IL-9R in diseased skin, and an association of blood IL-9 levels with clinical severity.. Little was known about the pathophysiological regulation of IL-9/IL-9R in AD skin. We asked whether IL-9R was expressed in epidermal keratinocytes; if so, what the functional outcome, cytokine production, and signaling pathway of IL-9/IL-9R in keratinocytes are.. We measured and compared the expression of IL-9R in skin from AD patients and controls by immunofluorescence. We also performed in vitro studies on the IL-9-treated primary keratinocytes, including flow cytometry for IL-9R expressions, Western blotting for mTOR, S6K, ERK, p38, and STAT3 activations, ELISA for cytokine levels, and immunofluorescence for STIM1.. We found that IL-9R was indeed expressed in keratinocytes but not in fibroblasts. Its expression in keratinocytes was enhanced by IL-4 but not by TGF-beta1. IL-9 induced a moderate production of IL-8 but not CXCL16, CCL22, TSLP, nor IL-33. IL-9 induced formation of STIM1-puncta. IL-9 induced ERK phosphorylation both dose- and time-dependently, but not mTOR, S6K, p38, or STAT3. Pretreatment with U0126 (ERK inhibitor) but not rapamycin (mTOR inhibitor) abrogated the IL-9-mediated IL-8 production. Blockage of STIM1 with BTP2 or SKF96265 abrogated ERK phosphorylation and IL-8 production induced by IL-9.. This study represents the first to show the regulation of the IL-9-STIM1-ERK-IL-8 axis in keratinocyte, and how the axis might play an important role in the pathophysiology of AD.

Topics: Animals; Cells, Cultured; Dermatitis, Atopic; Epidermal Cells; Extracellular Signal-Regulated MAP Kinases; Female; Humans; Interleukin-4; Interleukin-8; Interleukin-9; Keratinocytes; Membrane Proteins; Mice; Mice, Inbred C57BL; Neoplasm Proteins; Phosphorylation; Receptors, Interleukin-9; Stromal Interaction Molecule 1

Sanguisorba officinalis L. (SOL) is a perennial plant widely distributed in Asia, its roots are well-known as a traditional herbal medicine to treat burns, chronic intestinal infections, scalds, and inflammation in Korea. Also, the roots of SOL are used for treatment of many types of allergic skin diseases, including urticarial, eczema, and allergic dermatitis.. In this study we investigated the underlying mechanism of anti-inflammatory effect of an ethanol extract of SOL roots (ESOL).. The ability of ESOL to inhibit inflammatory skin disorder was tested in human keratinocyte HaCaT cells.. Viability test using MTT assay were used to determine non-cytotoxic concentrations of ESOL on HaCaT cells. ESOL-mediated inhibition of the tumor necrosis factor (TNF)-α/interferon (IFN)-γ-induced production of pro-inflammatory chemokines-such as macrophage-derived chemokine (MDC), regulated on activation, normal T-cell expressed and secreted (RANTES), interleukin (IL)-8, and thymus and activation regulated chemokine (TARC)-at the mRNA level was determined by real-time reverse transcription-polymerase chain reaction (RT-PCR). The ability of ESOL to reduce the expression of pro-inflammatory marker proteins was evaluated by Western blot analysis and immunocytochemistry.. ESOL reduced the production of MDC, RANTES, IL-8, and TARC in HaCaT cells stimulated with TNF-α/IFN-γ at both protein and mRNA levels. ESOL also suppressed the phosphorylation of signal transducer and activator of transcription (STAT)-1, extracellular signal-regulated kinase (ERK), and inhibited both nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor-alpha (IκB-α) degradation and the nuclear translocation of NF-κB/p65. ESOL exerts anti-inflammatory effects by suppressing the expression of TNF-α/IFN-γ-stimulated chemokines and pro-inflammatory molecules via a blockade NF-κB, STAT-1, and ERK activation.. Our results suggest the preventive potential of ESOL as a herbal medicine for the treatment of inflammatory skin diseases.

Topics: Cell Line; Chemokine CCL17; Chemokine CCL22; Chemokine CCL5; Chemokines; Dermatitis, Atopic; Extracellular Signal-Regulated MAP Kinases; Humans; I-kappa B Proteins; Inflammation; Interferon-gamma; Interleukin-8; Keratinocytes; NF-kappa B; NF-KappaB Inhibitor alpha; Phosphorylation; Phytotherapy; Plant Extracts; Sanguisorba; Skin; STAT1 Transcription Factor; Transcription Factor RelA; Tumor Necrosis Factor-alpha

CP001 is four traditional herbal medicine mixtures with anti-inflammatory properties. In this study, we investigated the effect of oral administration of CP001 ethanol extract on the 2,4-dinitrochlorobenzene- (DNCB-) induced AD mouse models. For that purpose, we observed the effects of oral administration of CP001 on skin inflammatory cell infiltration, skin mast cells, production of serum IgE, and expression of Th2 cytokine mRNA in the AD skin lesions of DNCB treated BALB/c mice. Histological analyses demonstrated that CP001 decreased dermis and epidermis thickening as well as dermal infiltration induced by inflammatory cells. In addition, CP001 decreased mast cell infiltration in count as well as dermal infiltration induced by inflammatory cells. In the skin lesions, mRNA expression of interleukin- (IL-) 4 and IL-13 was inhibited by CP001. CP001 also reduced the production of IgE level in mouse plasma. In addition, we investigated the effect of CP001 on the inflammatory allergic reaction using human mast cells (HMC-1). In HMC-1, cytokine production and mRNA levels of IL-4, IL-13, IL-6, and IL-8 were suppressed by CP001. Taken together, our results showed that oral administration of CP001 exerts beneficial effects in AD symptoms, suggesting that CP001 might be a useful candidate for the treatment of AD.

Topics: Animals; Chromatography, High Pressure Liquid; Dermatitis, Atopic; Dermis; Dinitrochlorobenzene; Enzyme-Linked Immunosorbent Assay; Epidermis; Immunoglobulin E; Interleukin-13; Interleukin-4; Interleukin-6; Interleukin-8; Male; Mice; Mice, Inbred BALB C; Real-Time Polymerase Chain Reaction; Th2 Cells

Allergy is characterized by eosinophilia and an increased susceptibility to microbial infection. Atopic dermatitis (AD) is typically associated with Staphylococcus aureus (SA) colonization. Some of the mechanisms by which SA and its exotoxins interact with eosinophils remain elusive. CD48, a glycosylphosphatidylinositol-anchored receptor belonging to the CD2 family, participates in mast cells-SA stimulating cross-talk, facilitates the formation of the mast cell/eosinophils effector unit and as expressed by eosinophils, mediates experimental asthma.. To investigate the role of CD48 expressed on human peripheral blood and mouse bone marrow-derived eosinophils (BMEos) in their interaction with heat-killed SA and its three exotoxins, Staphylococcal enterotoxin B (SEB), protein A (PtA) and peptidoglycan (PGN).. Eosinophils were obtained from human peripheral blood and BM of WT and CD48-/- mice. SA was heat killed and eosinophils-SA/exotoxins interactions were analyzed by confocal microscopy, adhesion and degranulation, cell viability, cytokine release and cell signalling. In addition, peritonitis was induced by SEB injection into CD48-/- and WT mice. CD48 expression was studied in AD patients' skin and as expressed on their leucocytes in the peripheral blood.. We provide evidence for the recognition and direct physical interaction between eosinophils and SA/exotoxins. Skin of AD patients showed a striking increase of eosinophil-associated CD48 expression while on peripheral blood leucocytes it was down-regulated. SA/exotoxins enhanced CD48 eosinophil expression, bound to CD48 and caused eosinophil activation and signal transduction. These effects were significantly decreased by blocking CD48 on human eosinophils or in BMEos from CD48-/- mice. We have also explored the role of CD48 in a SEB-induced peritonitis model in CD48-/- mice by evaluating inflammatory peritoneal cells, eosinophil numbers and activation.. These data demonstrate the important role of CD48 in SA/exotoxins-eosinophil activating interactions that can take place during allergic responses and indicate CD48 as a novel therapeutic target for allergy and especially of AD.

Topics: Animals; Antigens, CD; Bacterial Adhesion; Bone Marrow Cells; CD48 Antigen; Cell Degranulation; Dermatitis, Atopic; Enterotoxins; Eosinophils; Gene Expression; Humans; Interleukin-10; Interleukin-8; Leukocytes; Mice; Mice, Knockout; Peritonitis; Protein Binding; Signal Transduction; Skin; Staphylococcal Infections; Staphylococcus aureus

Atopic dermatitis is an inflammatory skin disease that is characterized by a reduced skin barrier function, reduced filaggrin (FLG) expression as well as increased colonization by Staphylococcus aureus.. This study focused on the possible involvement of FLG in epidermal colonization by S. aureus and/or whether it affects the epidermal defence mechanisms, including the expression of antimicrobial peptides (AMPs) and enzymes involved in stratum corneum barrier lipid synthesis. Furthermore, IL-31 has been shown to reduce FLG expression, but its effects on bacterial colonization and on the expression of AMPs and enzymes involved in the barrier lipid synthesis are not known.. We established N/TERT-based epidermal models (NEMs), after FLG knockdown (FLG-KD) and/or cultured with IL-31, that were colonized with S. aureus for 24 h.. Both FLG-KD and IL-31 supplementation resulted in significantly increased epidermal S. aureus colonization, as well as in an up-regulation of S. aureus-induced IL-8 expression. IL-31, but not FLG-KD, prevented S. aureus-induced up-regulation of mRNA expression for the AMPs human β-defensin 2 and -3 and RNAse7, whereas psoriasin expression remained unchanged. Furthermore, the S. aureus colonization induced changes in mRNA expression of ELOVL4 was not affected by FLG-KD, but was blocked by IL-31. Expression of SCD-1 and Gcase mRNA was reduced by IL-31, but not by FLG-KD.. This study shows that NEMs, with FLG-KD and/or cultured in the presence of IL-31, mimic the skin of patients with atopic dermatitis in several aspects, including enhanced bacterial colonization, increased inflammatory and reduced protective responses.

Topics: Adenosine Monophosphate; Animals; Cell Line; Dermatitis, Atopic; Disease Models, Animal; Epidermis; Filaggrin Proteins; Gene Expression Regulation; Gene Expression Regulation, Enzymologic; Gene Knockdown Techniques; Humans; Interleukin-8; Interleukins; Intermediate Filament Proteins; Lipids; Staphylococcal Infections; Staphylococcus aureus

The state of homeostasis links in the children with intestinal colic is represented by the following parameters and clinical characteristics. The data of investigated children's contingent with intestinal colic prevailed by following comorbidities: SARS--12 (18.18% ± 4.78%), protein-energy malnutrition--9 (12.85% ± 3.82%), pneumonia--6 (8.57% ± 3.57%), atopic dermatitis--7 (10.00% ±.3.57%). All children have a next complaints: flatulence (100%), in the 62 children (88.57% ± 3.82%) were identificated frequent regurgitation, in the 48 (80.33%)--hyperbilirubinemia. ALT levels were elevated in 25 children (41%) and 31 (51.66%) children had increased levels of AST. IL8 level were elevated in the 40 children (71.42%). The level of antibodies to elastase was greatly increased in all 56 (100%) children.

Topics: Alanine Transaminase; Aspartate Aminotransferases; Autoantibodies; Colic; Comorbidity; Dermatitis, Atopic; Female; Flatulence; Gastroesophageal Reflux; Humans; Hyperbilirubinemia; Infant; Infant, Newborn; Interleukin-8; Intestines; Male; Malnutrition; Pancreatic Elastase; Pneumonia, Bacterial; Severe Acute Respiratory Syndrome; Ukraine

Interleukin (IL)-17 is an important mediator of orthodontically induced inflammatory root resorption (OIIRR). However, its role in the dental pulp (DP) has not been studied. The aim of this study was to investigate, using an atopic dermatitis (AD) model, how IL-17 contributes to OIIRR in DP. Atopic dermatitis is the most common IL-17-associated allergic disease. Atopic dermatitis model mice (AD group) and wild-type mice (control group) were subjected to an excessive orthodontic force. The localization of T-helper (Th)17 cells, IL-17, IL-6, and keratinocyte chemoattractant (KC; an IL-8-related protein in rodents) were determined in DP. In addition, CD4+ T cells, including IL-17 production cells, were obtained from patients with AD and from healthy donors, and the effects of IL-17 on the production of IL-6 and IL-8 were investigated using a co-culture of CD4+ T cells with human dental pulp (hDP) cells stimulated with substance P (SP). Immunoreactivity for Th17 cells, IL-17, IL-6, and KC was increased in DP tissue subjected to orthodontic force in the AD group compared with DP tissue subjected to orthodontic force in the control group. The cells obtained from the AD patients displayed increased IL-6 and IL-8 production. These results suggest that IL-17 may aggravate OIIRR in DP.

Topics: Adolescent; Adult; Animals; CD4-Positive T-Lymphocytes; Cells, Cultured; Coculture Techniques; Dental Pulp; Dermatitis, Atopic; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin E; Interleukin-17; Interleukin-6; Interleukin-8; Male; Mice; Real-Time Polymerase Chain Reaction; Receptors, Interleukin-17; Root Resorption; Substance P; Th17 Cells; Tooth Movement Techniques

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

Atopic dermatitis (AD) is an inflammatory skin disease characterized by both acute and chronic eczema. Various markers are used to clinically evaluate the severity of AD. In order to identify a marker of local severity of AD, we measured IL-8, IL-18, vascular endothelial growth factor (VEGF), and transforming growth factor-α (TGF-α) levels in the stratum corneum (scIL-8, scIL-18, scVEGF and scTGF-α) and evaluated the correlation between the levels of these cytokines and the clinical severity scores of localized skin lesions.. Stratum corneum samples were collected from the skin lesions of 50 patients with AD using the tape-stripping technique, and the scIL-8, scIL-18, scVEGF and scTGF-α levels were evaluated using the ELISA method. The trans-epidermal water loss and skin water content of the lesions were also measured prior to tape stripping.. The levels of scIL-8, scIL-18, scVEGF and scTGF-α were significantly higher in patients with AD than in healthy controls. Additionally, the levels of scIL-8, scIL-18 and scVEGF significantly correlated with the severity of AD.. Among these cytokines, scIL-8 showed the highest correlation with the severity scores of lesions in AD as well as other parameters. Our results also suggest that measuring cytokines in the stratum corneum by using ELISA combined with tape stripping is a convenient method to evaluate the severity of skin lesions in AD.

Topics: Adolescent; Adult; Biomarkers; Child; Dermatitis, Atopic; Epidermis; Female; Humans; Interleukin-18; Interleukin-8; Male; Middle Aged; Severity of Illness Index; Transforming Growth Factor alpha; Vascular Endothelial Growth Factor A; Young Adult

IL-31 is a pruritogenic cytokine, and IL-33 is an alarmin for damaging inflammation. They together relate to the pathogenesis of atopic dermatitis (AD). Eosinophil infiltration into the inner dermal compartment is a predominant pathological feature of AD. We herein investigated the in vitro inflammatory effects of IL-31 and IL-33 on the activation of human eosinophils and dermal fibroblasts.. Receptors, adhesion molecules and signaling molecules were assessed by Western blot or flow cytometry. Chemokines and cytokine were quantitated by multiplex assay. Functional IL-31 receptor component IL-31RA, OSMR-β and IL-33 receptor component ST2 were constitutively expressed on the surface of eosinophils. Co-culture of eosinophils and fibroblasts significantly induced pro-inflammatory cytokine IL-6 and AD-related chemokines CXCL1, CXCL10, CCL2 and CCL5. Such inductions were further enhanced with IL-31 and IL-33 stimulation. IL-31 and IL-33 could significantly provoke the release of CXCL8 from eosinophils and fibroblasts, respectively, which was further enhanced upon co-culture. In co-culture, eosinophils and fibroblasts were the main source for the release of CCL5, and IL-6, CXCL1, CXCL8, CXCL10 and CCL2, respectively. Direct interaction between eosinophils and fibroblasts was required for CXCL1, CXCL10, CXCL8 and CCL5 release. Cell surface expression of intercellular adhesion molecule-1 on eosinophils and fibroblasts was up-regulated in co-culture upon IL-31 and IL-33 stimulation. The interaction between eosinophils and fibroblasts under IL-31 and IL-33 stimulation differentially activated extracellular signal-regulated kinase, c-Jun N-terminal kinase, p38 mitogen-activated protein kinase, nuclear factor-κB and phosphatidylinositol 3-kinase-Akt pathways. Using specific signaling molecule inhibitors, the differential induction of IL-31 and IL-33-mediated release of cytokines and chemokines such as IL-6 and CXCL8 from co-culture should be related to their distinct activation profile of intracellular signaling pathways.. The above findings suggest a crucial immunopathological role of IL-31 and IL-33 in AD through the activation of eosinophils-fibroblasts interaction via differential intracellular signaling mechanisms.

Topics: Blotting, Western; Cell Communication; Cells, Cultured; Chemokine CCL2; Chemokine CCL5; Chemokine CXCL1; Chemokine CXCL10; Coculture Techniques; Dermatitis, Atopic; Dermis; Eosinophils; Fibroblasts; Flow Cytometry; Humans; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Interleukin-6; Interleukin-8; Interleukins; Receptors, Cell Surface; Receptors, Interleukin; Recombinant Proteins

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

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

In this study, we synthesized a novel chemical, (E)-2-(3,4-dimethoxyphenyl)-4-oxo-4H-chromen-7-yl-3-(3,4-dimethoxyphenyl) acrylate (CSH) and investigated the effect of CSH on atopic dermatitis (AD) by evaluating the anti-inflammatory effect of CSH on immune cells in vitro and on atopic dermatitis-like lesions in vivo.. Human monocytic THP-1 cells and human eosinophilic EoL-1 cells were treated with house dust mite extract in the absence and presence of CSH. Nc/Nga mice were sensitized to 2,4-dinitrochlorobenzne (DNCB) for 5 weeks and then orally and dorsally administered with CSH or dexamethasone for 3 weeks.. CSH inhibited the secretion of monocyte chemotactic protein-1 (MCP-1), interleukin (IL)-6 and IL-8 due to house dust mite extract in THP-1 cells. CSH also suppressed the secretion of MCP-1 and IL-8 in EoL-1 cells. In vivo, the skin severity score decreased after CSH treatment as compared to the control group. CSH suppressed the inflammatory cell infiltration into the dermis and thickened the epidermis. CSH reduced serum IgE level as compared to the control group. The levels of IL-4, IL-5, IL-13 and eotaxin in mouse splenocytes increased after treatment with concanavalin A and the increase of the cytokines was decreased by pre-treatment with CSH. The inhibitory effects of CSH on atopic lesions of DNCB-treated Nc/Nga mice were similar to those of dexamethasone, despite differing degrees depending on results evaluated in this study.. These results may contribute to the development of a therapeutic drug for the treatment of AD.

Topics: Animals; Anti-Inflammatory Agents; Cell Line; Chemokine CCL2; Concanavalin A; Coumaric Acids; Coumarins; Cytokines; Dermatitis, Atopic; Dexamethasone; Disease Models, Animal; Eosinophils; Female; Humans; Immunoglobulin E; Interleukin-6; Interleukin-8; Mice; Monocytes; Pyroglyphidae; Severity of Illness Index; Spleen

Lipid rafts are cholesterol-rich cell signaling platforms, and their physiological role can be explored by cholesterol depletion. To characterize transcriptional changes ongoing after lipid raft disruption in epidermal keratinocytes, a cell type that synthesizes its cholesterol in situ, we performed whole-genome expression profiling. Microarray results show that over 3,000 genes are differentially regulated. In particular, IL-8, urokinase-like plasminogen activator receptor, and metalloproteinases are highly upregulated after cholesterol extraction. Quantitative reverse transcriptase PCR validation and protein release measurements demonstrate the physiological relevance of microarray data. Major enriched terms and functions, determined by Ingenuity Pathways Analysis, identify cholesterol biosynthesis as a major function, illustrating the specificity of keratinocyte response toward cholesterol depletion. Moreover, the inflammatory skin disorder atopic dermatitis (AD) is identified as the disease most closely associated with the profile of lipid raft-disrupted keratinocytes. This finding is confirmed in skin of AD patients, in whom transcript levels of major lipid raft target genes are similarly regulated in lesional atopic skin, compared with non-lesional and normal skin. Thus, lipid raft disruption evokes typical features of AD, thereby suggesting that lipid raft organization and signaling could be perturbed in atopic keratinocytes.

Topics: Biopsy; Cells, Cultured; Cholesterol; Dermatitis, Atopic; Epidermis; Gene Expression Profiling; Humans; Interleukin-8; Keratinocytes; Membrane Microdomains; Receptors, Urokinase Plasminogen Activator; Signal Transduction; Transcription, Genetic; Urokinase-Type Plasminogen Activator

Primary human keratinocytes and immortalized HaCaT cells were analysed for their capacity to bind purified staphylococcal protein A (SpA). Co-incubation with FITC-labelled SpA led to a dose-depending attachment. Pull-down experiments with cellular extracts revealed the TNFα receptor I (TNF RI) as binding partner on keratinocytes. Thus, we next looked for expression of this receptor in human epidermis and cultured keratinocytes. TNF RI is strongly expressed on all keratinocytes analysed, both at the mRNA and protein level and activation by SpA at optimal doses of 50-100 μg/ml resulted in the phosphorylation of the TNF RI downstream kinases MEK1/2, JNK1/2, and p38 subsequently leading to translocation of the p65 NF kappa B subunit and AP-1 into the nucleus. This translocation was then followed by increased expression of IL-8 and COX-2, two known NF kappa B-induced pro-inflammatory genes. To further test the relevance of our findings, we analysed in vitro production of over 100 strains isolated from atopic eczema showing that more than 85% of the tested strains produced extracellular SpA in substantial amounts. Thus, besides superantigens, haemolysins, and other cell wall components, Staphylococcus aureus exerts pro-inflammatory stimuli on human keratinocytes through the production of SpA signalling through TNF RI.

Topics: Active Transport, Cell Nucleus; Base Sequence; Cell Line; Cells, Cultured; Cyclooxygenase 2; Dermatitis, Atopic; Humans; In Vitro Techniques; Inflammation Mediators; Interleukin-8; Keratinocytes; Protein Binding; Receptors, Tumor Necrosis Factor, Type I; RNA, Messenger; Staphylococcal Protein A; Staphylococcus aureus; Transcription Factor AP-1; Transcription Factor RelA; Transcription, Genetic

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

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

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

Bacterial infection with Staphylococcus aureus is a known trigger for worsening of atopic dermatitis (AD); the exact mechanisms by which bacterial infection worsens dermatitis are unknown.. We sought to characterize the amounts of the biologically active bacterial lipoprotein lipoteichoic acid (LTA) in infected AD lesions.. Eighty-nine children with clinically impetiginized lesions of AD were enrolled in this study. A lesion was graded clinically by using the Eczema Area and Severity Index (EASI), wash fluid obtained from the lesion for quantitative bacterial culture, and measurement of LTA and cytokines. The staphylococcal isolate was tested for antibiotic susceptibilities. The patients were treated with a regimen that included topical corticosteroids and systemic antibiotics, and the lesion was reanalyzed after 2 weeks.. S aureus was identified in 79 of 89 children enrolled in the study. The bacterial colony-forming unit (CFU) counts correlated with the EASI lesional score (P = .04). LTA levels as high as 9.8 mug/mL were measured in the wash fluid samples, and the amounts correlated with the lesional EASI scores (P = .01) and S aureus CFU (P < .001). Approximately 30% of clinically impetiginized AD lesions contained greater than 1 mug/mL LTA, amounts that exert effects on various cell types in vitro. Moreover, injection of skin tissue ex vivo with amounts of LTA found in AD lesions resulted in epidermal cytokine gene expression.. Pharmacologic levels of LTA are found in many infected atopic dermatitis lesions.

Topics: Child; Child, Preschool; Colony Count, Microbial; Dermatitis, Atopic; Eczema; Humans; Infant; Interleukin-8; Lipopolysaccharides; Severity of Illness Index; Skin; Staphylococcal Skin Infections; Staphylococcus aureus; Teichoic Acids; Tumor Necrosis Factor-alpha

Topics: Acute-Phase Proteins; beta-Defensins; Chemokine CCL20; Cytokines; Dermatitis, Atopic; Gene Expression; Gene Expression Profiling; Humans; Immunohistochemistry; Interleukin-17; Interleukin-8; Keratinocytes; Lipocalin-2; Lipocalins; Proto-Oncogene Proteins; Receptors, Interleukin-17; Reverse Transcriptase Polymerase Chain Reaction; T-Lymphocytes, Helper-Inducer

Innate immune responses involve the production of antimicrobial peptides (AMPs), chemokines, and cytokines. We report here the identification of B-cell leukemia (Bcl)-3 as a modulator of innate immune signaling in keratinocytes. In this study, it is shown that Bcl-3 is inducible by the Th2 cytokines IL-4 and IL-13 and is overexpressed in lesional skin of atopic dermatitis (AD) patients. Bcl-3 was shown to be important to cutaneous innate immune responses as silencing of Bcl-3 by small-interfering RNA (siRNA) reversed the downregulatory effect of IL-4 on the HBD3 expression. Bcl-3 silencing enhanced vitamin D3 (1,25D3)-induced gene expression of cathelicidin AMP in keratinocytes, suggesting a negative regulatory function on cathelicidin transcription. Furthermore, 1,25D3 suppressed Bcl-3 expression in vitro and in vivo. This study identified Bcl-3 as an important modulator of cutaneous innate immune responses and its possible therapeutic role in AD.

Topics: Amino Acid Sequence; Antimicrobial Cationic Peptides; B-Cell Lymphoma 3 Protein; Cathelicidins; Cells, Cultured; Cholecalciferol; Dermatitis, Atopic; Humans; Immunity, Innate; Interleukin-13; Interleukin-4; Interleukin-6; Interleukin-8; Keratinocytes; Molecular Sequence Data; NF-kappa B p50 Subunit; Proto-Oncogene Proteins; Transcription Factors

House dust mites produce serine and cysteine proteases. Mite-derived proteases have been suggested to be involved in the pathogenesis of allergies; however, whether mite-derived serine protease activity can stimulate keratinocytes remains unknown.. We examined the activation of primary human keratinocytes by serine protease-rich extract of whole mite culture and compared with that by recombinant group 1 allergens (rDer f 1 and rDer p 1), which exclusively exhibit cysteine protease activity.. Protease activity of whole mite culture extract (WCE), rDer f 1 and rDer p 1 induced the release of IL-8 and granulocyte-macrophage colony-stimulating factor. Protease activity of WCEs induced a significant upregulation of their mRNA expression but rDer f 1 had much less effect. Protease activity of the WCE stimulated intracellular Ca(2+) mobilization but rDer f 1 and rDer p 1 did not. The mobilization induced by agonists for the human protease-activated receptor (PAR)-2, an agonist peptide or trypsin, was diminished by pre-incubation of keratinocytes with WCE. rDer f 1 inefficiently cleaved a synthetic N-terminal peptide of PAR-2 at different sites from trypsin, but the resultant peptides did not stimulate the release of interleukin-8.. The results suggest that mite-derived serine protease activity may contribute to the pathogenesis of atopic dermatitis by activating keratinocytes via PAR-2 activation but cysteine protease activity of Der f 1 and Der p 1 acts via another mechanism.

Topics: Animals; Antigens, Dermatophagoides; Arthropod Proteins; Calcium; Cells, Cultured; Cysteine Endopeptidases; Dermatitis, Atopic; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Keratinocytes; Peptides; Pyroglyphidae; Receptor, PAR-2; Recombinant Proteins; RNA, Messenger; Serine Proteases

Keratinocytes may play an important role in the pathogenesis of skin disease in atopic dermatitis. Diarylheptanoids such as oregonin and hirstanonol are demonstrated to have anti-inflammatory and anti-oxidant effects. The present study was to investigate the effect of hirsutenone, one of the diarylheptanoids, against tumor necrosis factor (TNF)-alpha-stimulated responses in human keratinocytes. Hirsutenone attenuated the TNF-alpha-induced production of cytokine IL-8, prostaglandin E(2) and chemokine CCL27, and the formation of reactive oxygen/nitrogen species in keratinocytes. Immunosuppressants (dexamethasone and cyclosporin A) inhibited the TNF-alpha-elicited formation of IL-8, prostaglandin E(2) and CCL27, but did not affect formation of reactive species. Bay 11-7085 (an inhibitor of NF-kappaB activation) and anti-oxidant N-acetylcysteine attenuated the TNF-alpha-induced formation of inflammatory mediators and reactive species. Hirsutenone, dexamethasone, cyclosporin A and Bay 11-7085 inhibited the TNF-alpha-induced phosphorylation of inhibitory kappaB and the activation of nuclear factor (NF)-kappaB. The results show that hirsutenone seems to reduce the TNF-alpha-stimulated production of inflammatory mediators in keratinocytes by suppressing the activation of NF-kappaB that may be mediated by reactive oxygen species. The findings suggest that hirsutenone may exert an inhibitory effect against the pro-inflammatory mediator-induced skin disease.

Topics: Alnus; Catechols; Cell Survival; Cells, Cultured; Chemokine CCL27; Dermatitis, Atopic; Diarylheptanoids; Dinoprostone; Gene Expression Regulation; Humans; Interleukin-8; Keratinocytes; NF-kappa B; Phytotherapy; Plant Extracts; Tumor Necrosis Factor-alpha

Although exposure to mite allergens is an important risk factor for the production of IgE and is associated with various allergic diseases, there has been uncertainty as to the route of exposure by which sensitization occurs. Cystatin A is a skin-derived dominant inhibitor against proteolytic activity of major mite allergens, Der f 1 and Der p 1, and blocks the upregulation of IL-8 release from human keratinocytes stimulated with the allergens. We analyzed whether the stimulation of keratinocytes with the allergens upregulates the release of granulocyte-macrophage colony-stimulating factor (GM-CSF), which has many actions relevant to allergic diseases including atopic dermatitis, and if so, whether cystatin A can block this process.. Normal human keratinocytes and the human keratinocyte cell line HaCaT were stimulated with recombinant group 1 allergens in the absence or presence of cystatin A.. Stimulation with the recombinant allergens upregulated the release of GM-CSF from normal human keratinocytes in a culture with high calcium concentration and HaCaT cells, which could be inhibited by the addition of cystatin A. The allergens exhibiting proteolytic activity did not digest cystatin A. Proteolytic activity of recombinant Der f 1 was partially regenerated after incubation with keratinocytes even without preactivation by L-cysteine.. Proteolytic activity of recombinant Der f 1 and Der p 1 upregulates GM-CSF and IL-8 release from keratinocytes in vitro, suggesting possible contributions to sensitization through the skin and the perpetuation of atopic dermatitis, as well as a homeostatic role for cystatin A against inflammation of the skin.

Topics: Animals; Antigens, Dermatophagoides; Arthropod Proteins; Cell Line; Cystatins; Cysteine Endopeptidases; Dermatitis, Atopic; Electrophoresis, Polyacrylamide Gel; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-8; Keratinocytes; Protease Inhibitors; Pyroglyphidae; Up-Regulation

Cytokines play an important role in skin inflammation.. We determined whether polymorphisms in cytokine genes contribute to the occurrence of occupational chronic irritant contact dermatitis (CICD).. In a case-control study, 9 polymorphisms in the genes coding for interleukin (IL)-1 alpha, IL-1 beta, IL-8, IL-10, and tumour necrosis factor (TNF)-alpha were determined in 197 patients with CICD. 217 apprentices in vocational training for high-risk occupations for CICD served as controls.. For all polymorphisms, no differences in genotype distributions were found between patients and controls. However, in patients with self-reported low levels of wet work and irritant exposure, more TNFA -308 variant genotypes (G/A and A/A) were present compared with those exposed to higher levels or controls, which indicates a TNFA-induced increase of susceptibility. In patients with TNFA -308 variant genotypes, the prevalence of flexural eczema was higher (48% and 57%) compared with that in patients presented with wild-type genotype (30%). Regarding IL1A -889, prevalence of symptoms of dermatitis was lower in apprentices with T/T or C/T genotype (32% and 36%) compared with wild-type genotype (54%, C/C). This indicates a protective effect of these variant alleles in acquiring hand dermatitis.. This study provides evidence that some genetic variations alter susceptibility to (chronic) dermatitis. Knowledge of the impact of genetic differences on the risk of CICD is essential in predictive testing of individuals at risk.

Topics: Adolescent; Adult; Case-Control Studies; Chronic Disease; Dermatitis, Atopic; Dermatitis, Irritant; Dermatitis, Occupational; DNA; Female; Gene Frequency; Genetic Predisposition to Disease; Genotype; Hand Dermatoses; Humans; Interleukin-10; Interleukin-1alpha; Interleukin-1beta; Interleukin-8; Logistic Models; Male; Middle Aged; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Tumor Necrosis Factor-alpha

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

High-throughput technologies, including DNA-chip array, have been used to search for the genes that are dysregulated in human diseases. The atopic dermatitis (AD)-associated genes are gradually being reported; however, the differentially altered gene expression profiles of atopic fibroblasts have not been well elucidated.. We wanted to gain more insights into AD and to find candidate genes, especially in regards to the role of fibroblasts in the pathogenesis of AD.. cDNA microarray (8K) profiling of the primary cultured AD patients-derived fibroblasts was conducted by a pooling method of the recruited 22 normal controls, the 10 extrinsic type (ADe) patients and the 10 intrinsic type (ADi) patients. SAM analysis of the microarray results (2-fold cut-off) was conducted to select the candidate genes. Quantification by real-time PCRs confirmed the array data in the randomized paired samples (normal vs. ADe n=10; normal vs. ADi n=10).. We listed the 22 up-regulated and 95 down-regulated genes in the AD fibroblasts. Real-time PCR results showed that several genes such as hyaluronan synthase 2 (HAS2), TNF-alpha-induced protein 6 (TNFAIP6) and IL-8 were matched with the array results with statistical significance.. These results suggest gene expression profiles that are associated with AD and this implied that fibroblasts may play important roles in the AD pathogenesis. We provided new insights into three candidate genes such as HAS2, TNFAIP6 and IL-8 with respect to their involvement in AD disease.

Topics: Adult; Aldehyde Dehydrogenase; Aldehyde Dehydrogenase 1 Family; Amino Acid Oxidoreductases; Brain-Derived Neurotrophic Factor; Cell Adhesion Molecules; Cells, Cultured; Chemokines, CC; Dermatitis, Atopic; DNA, Complementary; Down-Regulation; Extracellular Matrix; Fibroblasts; Gene Expression Profiling; Glucuronosyltransferase; Humans; Hyaluronan Synthases; Interleukin-8; Male; Oligonucleotide Array Sequence Analysis; Polymerase Chain Reaction; Proteoglycans; Retinal Dehydrogenase; Up-Regulation; Vesicular Transport Proteins

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

Sargassum hemiphyllum (SH) has long been used in Korean folk medicine for the therapeutic treatment of various allergic diseases. The effects of SH in previous experimental models, however, have been inconclusive. We studied the effects of methanol extract of SH on mast cells. Our experiments showed that SH significantly inhibited compound 48/80-induced histamine and beta-hexosaminidase release from rat peritoneal mast cells. SH inhibited interleukin (IL)-8 and tumor necrosis factor (TNF)-alpha release induced by phorbol 12-myristate 13-acetate and A23187 from HMC-1, and it also showed an inhibitory effect on the anti-dinitrophenyl IgE antibody-induced passive cutaneous anaphylaxis reaction. In addition, SH inhibited the increase of TNF-alpha-induced NF-kappaB protein levels, transcription factor of TNF-alpha from 293T cells. A period of 48 h exposure to SH had little effect on HMC-1 cell viability. Our results suggest that SH has an inhibitory effect on the atopic allergic reaction and thus this may be useful in the treatment of allergic inflammatory diseases, such as atopic dermatitis.

Topics: Animals; Anti-Allergic Agents; beta-N-Acetylhexosaminidases; Cell Line; Cell Line, Tumor; Dermatitis, Atopic; Histamine Release; Humans; Inflammation Mediators; Interleukin-8; Korea; Mast Cells; Medicine, East Asian Traditional; Mice; Mice, Inbred ICR; NF-kappa B; Phytotherapy; Rats; Rats, Wistar; Sargassum; Tumor Necrosis Factor-alpha

Carpopeltis affinis Okamura (CA, Halymeniaceae) has long been used as therapeutics for various allergic diseases in Korea. The precise effects of CA in experimental models, however, have remained unknown. We studied the effects of a methanol extract of CA on atopic allergic reaction. Histamine content was measured by the o-phthalaldehyde spectrofluorometric procedure. Cytokines were measured by a modified enzyme-linked immunosorbent assay. Cytotoxicity was determined by the 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. CA significantly inhibited the histamine release and beta-hexosaminidase release from rat peritoneal mast cells. CA also inhibited interleukin-8 and tumor necrosis factor-alpha secretion from the phorbol 12-myristate 13-acetate and A23187-induced HMC-1 cells (human mast cell line). 48 h exposure to CA (1.0, 10, and 100 microg/ml) had little effect on HMC-1 cell viability. Our results suggest that CA has an inhibitory effect on mast cell-dependent allergic reaction and thus may be useful in the treatment of atopic dermatitis.

Topics: Animals; beta-N-Acetylhexosaminidases; Cell Survival; Dermatitis, Atopic; Histamine Release; Interleukin-8; Korea; Magnetic Resonance Spectroscopy; Mast Cells; Phytotherapy; Plant Extracts; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

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

The T helper type-2 (Th2)-dominated situation can be observed in allergic diseases such as asthma or atopic dermatitis. A reduced ability to produce IL-12, which is a key cytokine for the induction of Th1 responses, has been proposed to lead to aberrant Th2 development in these disease conditions.. This study was intended to examine how IL-12-producing ability might associate with allergic diseases as a function of age.. IL-12 production by monocytes at various ages was assessed in patients with bronchial asthma and/or atopic dermatitis (n = 100) in comparison with non-allergic control subjects (n = 144). Whole blood cells were stimulated with lipopolysaccharide (LPS) after priming with IFN-gamma, then intracellular cytokine expression of IL-12 and IL-8 as a control cytokine of CD14-positive cells was assessed by flow cytometric analysis.. In the control subjects, the ability of monocytes to produce IL-12 was negligible at birth and gradually increased with advancing age, whereas IL-8 production was intense throughout the human life. At more than 7 years of age, IL-12 production of patients with allergic diseases was significantly lower compared with that of control subjects. The unexpected finding was that infants and children below 6 years of age with allergic diseases tended to produce more IL-12 compared with age-matched controls. In this young group, it was noted that enhanced IL-12 production by monocytes was especially observed in allergic patients with specific IgE antibodies against some food allergens. Significant inverse relationships between serum IgE levels and IL-12-producing ability were found in the teenage and adult groups, but not in the younger children.. IL-12 appeared to play different roles in the pathogenesis of allergic diseases between younger and older ages.

Topics: Adolescent; Adult; Aging; Asthma; Child; Child, Preschool; Dermatitis, Atopic; Female; Flow Cytometry; Humans; Hypersensitivity; Infant; Infant, Newborn; Interferon-gamma; Interleukin-12; Interleukin-8; Lipopolysaccharide Receptors; Lipopolysaccharides; Male; T-Lymphocytes; Th1 Cells; Th2 Cells

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

Chronic skin colonization with Staphylococcus aureus is a characteristic feature of atopic eczema (AE), and about 60% of S. aureus strains isolated from the skin of patients with AE secrete enterotoxins. Furthermore, IgE antibodies to S. aureus enterotoxins have been identified in 78% of patients with AE.. To examine the S. aureus enterotoxin-induced histamine and leukotriene release of basophils from patients with AE.. Peripheral blood basophils from patients with AE were stimulated with the staphylococcal enterotoxins A, B, D, E and toxic shock syndrome toxin-1. Additionally, priming experiments were performed with interleukin (IL)-3, IL-8 and granulocyte/macrophage colony-stimulating factor followed by stimulation with S. aureus enterotoxins.. In patients with AE, basophils secreted significantly higher amounts of histamine and leukotriene C4 (LTC4) than in healthy controls. The priming experiments showed additional histamine and LTC4 release in the group of AE patients.. Histamine and leukotriene generation from atopic basophils stimulated with staphylococcal enterotoxins may indicate a role for these toxins as possible allergens in at least a subgroup of patients with AE.

Topics: Bacterial Toxins; Basophils; Case-Control Studies; Dermatitis, Atopic; Enterotoxins; Enzyme-Linked Immunosorbent Assay; Exotoxins; Granulocyte-Macrophage Colony-Stimulating Factor; Histamine Release; Humans; Interleukin-3; Interleukin-8; Leukotriene C4; Staphylococcal Protein A; Staphylococcus aureus; Statistics, Nonparametric; Superantigens

The release of cytokines [interleukin-6 (IL-6), IL-8 and tumor necrosis factor-alpha (TNF-alpha)] by skin cells is involved in the pathogenesis of atopic dermatitis (AD).. To evaluate the effect of low-dose cyclosporin A (CyA) on clinical symptoms and cytokine secretion in severe pediatric AD.. Ten children with severe AD (SCORAD index >50) were treated for 8 weeks with CyA. The initial dose of 2.5 mg/kg/day was titrated to a maximum of 5 mg/kg/day until a SCORAD reduction of >or =35% was achieved ("treatment response"). After stopping CyA all patients entered a 4-week follow-up period. Cytokine secretion (IL-6, IL-8 and TNF-alpha) from patients' PBMC was assessed by ELISA before and after CyA treatment and was compared with 18 healthy nonatopic controls. Only the data of patients, who responded to CyA and did not experience a relapse during the follow-up period, were evaluated for this paper.. Seven patients responded to CyA without relapse during the follow-up period. The median SCORAD index in these patients improved from 71 at baseline to 22 after CyA treatment (p < 0.001). AD patients' PBMC produced more IL-6, IL-8 and TNF-alpha than PBMC of controls. Suppression of IL-6 (p < 0.05) and IL-8 (p < 0.05) production was observed after CyA treatment. TNF-alpha levels were unchanged by CyA in all patients.. The reduction in severity of pediatric AD with CyA is associated with decreased production of IL-6 and IL-8, but not TNF-alpha by PBMC.

Topics: Adolescent; Child; Child, Preschool; Cyclosporine; Dermatitis, Atopic; Emulsions; Female; Humans; Immunosuppressive Agents; Infant; Interleukin-6; Interleukin-8; Male; Prospective Studies; Tumor Necrosis Factor-alpha

IL-18 has been found to be an IFN-gamma-inducing factor that plays an important role in T(H)1 cell activation. Recently, IL-18 has also been found to enhance a T(H)2 cellular response in a specific setting.. The aim of this study was to elucidate the role of monocytes and soluble factors, with special focus on IL-18, in the pathogenesis of atopic dermatitis (AD).. The release of cytokines from PBMCs and purified monocytes was measured through use of ELISA; mRNA expression was evaluated by RT-PCR. The results from patients with AD were compared with those from healthy controls.. IL-18 secretion was reduced in both unstimulated and lipopolysaccharide-stimulated monocytes from patients with AD. The mRNA expression of IL-18 and IL-1 beta-converting enzyme was significantly reduced in unstimulated monocytes from patients with AD (P <.03 and P <.01, respectively). Patients with AD had an elevated secretion of prostaglandin E(2) (PGE(2)) from unstimulated PBMCs (P <.001). The anti-PGE(2) antibody reversed the suppressive effect of PGE(2) on IL-18 secretion in unstimulated PBMCs from patients with AD.. Decreased IL-18 production, together with a significantly reduced IL-18 and ICE mRNA expression in unstimulated monocytes and elevated PGE(2) secretion from PBMCs, was associated with the pathogenesis of AD.

Topics: Adolescent; Adult; Caspase 1; Cytokines; Dermatitis, Atopic; Dinoprostone; Female; Humans; Interleukin-1; Interleukin-10; Interleukin-18; Interleukin-8; Lipopolysaccharide Receptors; Male; Middle Aged; Monocytes; RNA, Messenger

IL-8 and related Glu-Leu-Arg (ELR+) CXC chemokines are potent chemoattractants for neutrophils but not for monocytes. IL-13 and IL-4 strongly increased CXCR1 and CXCR2 chemokine receptor expression in human monocytes, macrophages, and dendritic cells. The effect was receptor- and cell type-selective, in that CCRs were not increased and no augmentation was seen in neutrophils. The effect was rapid, starting at 4 h, and concentration dependent (EC50 = 6.2 and 8.3 ng/ml for CXCR1 and CXCR2, respectively) and caused by new transcriptional activity. IL-13/IL-4-treated monocytes showed increased CXCR1 and CXCR2 membrane expression. IL-8 and related ELR+ chemokines were potent and effective chemotactic agents for IL-13/IL-4-treated monocytes, but not for untreated mononuclear phagocytes, with activity comparable to that of reference monocyte attractants, such as MCP-1. In the same cells, IL-8 also caused superoxide release. Macrophages and dendritic cells present in biopsies from Omenn's syndrome and atopic dermatitis patients, two Th2 skewed pathologies, expressed IL-8 receptors by immunohistochemistry. These results show that IL-13 and IL-4 convert IL-8 and related ELR+ chemokines, prototypic neutrophil attractants, into monocyte chemotactic agonists, by up-regulating receptor expression. Therefore, IL-8 and related chemokines may contribute to the accumulation and positioning of mononuclear phagocytes in Th2-dominated responses.

Topics: Antigens, CD; Blotting, Northern; Chemotaxis, Leukocyte; Dermatitis, Atopic; Free Radicals; Humans; Interleukin-13; Interleukin-4; Interleukin-8; Monocytes; Reactive Oxygen Species; Receptors, Chemokine; Receptors, Interleukin; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Respiratory Burst; Severe Combined Immunodeficiency

Inflammatory mechanisms play an important role in the pathogenesis and the expansion of the skin lesion in AD. The serum levels of NO product, IL-8, RANTES and eotaxin, which are considered to an index of the inflammatory response, were measured in infants diagnosed as AD. The serum levels of NO product, RANTES and eotaxin were higher in the infantile AD patients with the systemic skin lesion compared with controls. The higher levels of NO product were shown with the expansion of the skin lesion. In AD patients, the serum levels of NO product were significantly correlated with the serum levels of eotaxin (r = 0.615, p < 0.001). These results suggest that NO product bears an important function in the allergic inflammation, which is concerned with the lesion expansion of the infantile AD patients and may be an index of the allergic inflammation.

Topics: Chemokine CCL11; Chemokine CCL5; Chemokines, CC; Chemotactic Factors, Eosinophil; Cytokines; Dermatitis, Atopic; Female; Humans; Infant; Interleukin-8; Male; Nitric Oxide

Topics: Biomarkers; Dermatitis, Atopic; E-Selectin; Female; Humans; Immunoglobulin E; Infant; Intercellular Adhesion Molecule-1; Interleukin-8; Male; Vascular Cell Adhesion Molecule-1

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

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

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

Interactions between keratinocytes and mononuclear cells via cytokines and adhesion molecules are thought to play a crucial part in inflammatory skin diseases. The cytokine-mediated effects of peripheral blood mononuclear cells (PBMC) from patients with atopic eczema (AE) and healthy individuals on keratinocytes (HaCaT) were investigated in vitro. A new coculture model (Transwell system) which consists of a lower and an upper compartment, which are separated by a polycarbonate-treated membrane, was established. 3[H]thymidine incorporation of keratinocytes and lymphocytes, as well as IL-6, IL-8 and IFN-gamma synthesis, were measured. Keratinocyte proliferation was significantly enhanced in the presence of PBMC from patients with AE. In contrast, PBMC from normal donors did not enhance HaCaT cell proliferation when they were cocultured. Lymphocytes from patients with AE showed a significantly enhanced proliferation after coculture with keratinocytes. However, PBMC from normal donors did not proliferate in the presence of HaCaT cells. Keratinocyte supernatants incubated with PBMC from either atopic or normal volunteers induced a suppression of lymphocyte 3[H]thymidine incorporation. In supernatants from cocultures of PBMC from patients with AE and keratinocytes, significantly enhanced amounts of IL-6 and IL-8, compared with normal donor's lymphocytes and HaCaT cells, were measured. No differences in IFN gamma production were observed. When PBMC were cultured without HaCaT cells, supernatants contained equal levels of IL-6, IL-8 and IFN-gamma in normal donors and in patients with AE. Interestingly, HaCaT cells spontaneously secrete measurable amounts of IL-6, IL-8 and IFN-gamma. Blocking experiments with neutralizing antibodies against these interleukins showed a complete inhibition of keratinocyte proliferation when PBMC from normal donors were used whereas the proliferative potency of PBMC supernatants from patients with AE on keratinocytes remained. Our data indicate that (i) PBMC from patients with AE stimulate keratinocyte proliferation via soluble factor(s) that are different from IL-6, IL-8 and IFN-gamma; (ii) probably, HaCaT cells spontaneously produce lymphocyte/monocyte inhibitory soluble factors and IL-6, IL-8 as well as IFN-gamma; and (iii) secretion and/or activity of keratinocyte-derived inhibitory mediators is regulated via cytokines of PBMC infiltrating inflammatory skin.

Topics: Antibodies, Monoclonal; Cell Culture Techniques; Cell Division; Cell Line; Coculture Techniques; Cytokines; Dermatitis, Atopic; Humans; Interferon-gamma; Interleukin-6; Interleukin-8; Keratinocytes; Leukocytes, Mononuclear; Thymidine

Plasma interleukin-8 (IL-8) concentrations were measured in patients with atopic dermatitis. Plasma IL-8 was not detected in 25 controls (0/25), in allergic rhinitis (0/20), or in bronchial asthma during remission (0/13), while low concentrations of IL-8 were detectable in a few patients with urticaria (1/19), contact dermatitis (4/17), and bronchial asthma at the time of attack (6/16). In contrast, IL-8 was detectable in most cases of atopic dermatitis (41/52). Moreover, IL-8 concentrations were significantly higher in severe than in mild or moderate atopic dermatitis. IL-8 concentrations decreased as atopic dermatitis was improved by treatment, and IgE production in vitro was also decreased while serum IgE concentrations remained unchanged. IL-8 measurement may be a useful tool for the study of the pathogenesis and clinical course of atopic dermatitis.

Topics: Adolescent; Asthma; Cells, Cultured; Child; Child, Preschool; Dermatitis, Atopic; Dermatitis, Contact; Female; Humans; Immunoglobulin E; Infant; Interleukin-1; Interleukin-6; Interleukin-8; Male; Rhinitis; Urticaria

Eosinophil granular protein deposits have been demonstrated in lesional atopic dermatitis skin. This suggests active tissue infiltration of eosinophils. To find an explanation for the tissue influx of eosinophils, eosinophil migration was studied in vitro by means of a microchemotaxis assay. Eosinophils from the circulation of patients with atopic dermatitis showed an altered capacity to respond to chemotactic stimuli in vitro compared with eosinophils from healthy donors. Eosinophils from patients with atopic dermatitis had significantly increased migratory responses toward dose ranges of N-formyl-methionyl-leucyl-phenylalanine, neutrophil-activating factor, platelet-activating factor, and platelet factor 4. Eosinophils from normal individuals did not respond to N-formyl-methionyl-leucyl-phenylalanine and neutrophil-activating factor and responded only slightly to platelet factor 4. The migratory responses toward tumor necrosis factor-alpha and complement factor C5a were identical in both groups. Interleukin-5, an eosinophil-selective cytokine, is a strong modulator of the migratory responses to these chemotaxins in eosinophils from normal donors. A migratory response toward N-formyl-methionyl-leucyl-phenylalanine and neutrophil-activating factor was induced by interleukin-5, whereas the migratory response toward platelet-activating factor and platelet factor 4 was markedly potentiated. In contrast, the response to complement fragment C5a was only slightly influenced. Our findings indicate that the increased migratory responsiveness of eosinophils from patients with atopic dermatitis to various chemotaxins reflects in vivo "priming" of eosinophils, presumably by circulating cytokines such as interleukin-5. This in vivo "priming" is not optimal because it can be further potentiated by renewed contact with interleukin-5.

Topics: Adolescent; Adult; Cell Movement; Chemotaxis, Leukocyte; Complement C5a; Dermatitis, Atopic; Eosinophils; Female; Humans; Interleukin-5; Interleukin-8; Male; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Platelet Activating Factor; Platelet Factor 4

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

The neutrophil activating peptide NAP-1/IL-8 has in the past been shown to be secreted by diverse cell-types involved in inflammatory processes. Furthermore, potent biological effects on both neutrophilic granulocytes and lymphocytes enforce its role in inflammation. Recently, immunohistochemical studies using monoclonal anti-NAP-1/IL-8 antibodies have been performed on dermal inflammatory conditions like psoriasis vulgaris. These have demonstrated epidermal IL-8 immunoreactivity in a pattern inversely related to the degree of inflammatory infiltration. Based on these results, in the present study biopsies from patients with contact eczema as well as atopic dermatitis were examined. The same patterns of immunoreactivity were found with either homogeneous epidermal staining, focally negative staining or overall decreased or even absent staining. As in psoriasis, these patterns were related to the degree of inflammatory infiltration. These results prove NAP-1/IL-8 to be involved not only in psoriasis vulgaris, but more likely to be a marker of different inflammatory processes. Future work will have to examine the kinetics as well as stimuli causing these effect.

Topics: Biopsy; Dermatitis, Atopic; Dermatitis, Contact; Humans; Immunoenzyme Techniques; Interleukin-8

The influence of the receptor-specific stimuli interleukin-3 (IL-3), interleukin-8 (IL-8), C5a and formyl-methionyl-leucyl-phenylalanine (FMLP) on the generation of arachidonic acid-derived inflammatory mediators from neutrophils (PMN) has been studied in patients with atopic dermatitis (AD) as well as in healthy, non-atopic volunteers. The release of leukotriene (LT)B4, the omega-oxidation products 20-COOH- and 20-OH-LTB4 and the cysteinyleukotriene LTC4 were measured by reverse-phase HPLC and radioimmunoassay. The incubation of neutrophils with these stimuli led to a significantly higher release of LTB4 and LTC4 in the AD group. The spontaneous leukotriene generation of PMN from patients with AD was on average threefold higher compared to the control group. C5a stimulated the release of LTB4 and its metabolites from atopic cells up to 9 ng in contrast to low amounts from non-atopic cells. Furthermore, FMLP distinctly enhanced the leukotriene release of neutrophils from patients with AD compared to unstimulated cells and to cells of normal donors. IL-3 and IL-8 also significantly stimulated the generation of LTB4 and LTC4 of PMN from atopic patients. Our data emphasize that neutrophils may play an important role in the pathogenesis of AD by an increased responsiveness to receptor-specific stimuli and further suggest that IL-3 and IL-8 influence the acute and chronic inflammatory reactions in patients with AD.

Topics: Cells, Cultured; Complement C5a; Dermatitis, Atopic; Humans; Interleukin-3; Interleukin-8; Interleukins; Leukotrienes; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils