interleukin-8 and Skin-Diseases

This thesis discusses the phenotypic characteristics of different inflammatory dermatological diseases and sets this into context with the specific chemotactic ability of different cytokines. It further discusses the biological properties of different chemotactic cytokines and their relevance in certain inflammatory diseases. The term chemotaxis was introduced in 1884 by Pfeffer, who described it as directional migration of leukocytes along a gradient. Regular studies of chemotaxis were, however, not possible until 1962 when Boyden developed the chemotaxis chamber technique. This test has since then been improved, and it is now possible to define and characterize chemoattractants and examine the special chemotactic behavior of leukocytes. We investigated T lymphocyte responses towards different chemoattractants using a modified Boyden chamber technique and found that approximately 50% of normal individuals have cells which respond whereas T-cells from the remaining persons did not respond. We therefore chose human T lymphocytic cell lines as target cells for chemotaxis screening to avoid inter-individual variations among donors. T lymphocytic infiltrates dominated by CD4+, CD45R0+ memory T cells are characteristic for many dermatological inflammatory diseases. We have therefore performed experiments to evaluate whether an earlier described epidermal lymphocyte chemotactic factor (ELCF) from skin overlying a tuberculin skin reaction in addition with other cytokines specifically attracts different subsets of lymphocytes. ELCF which probably reflects a mixture of different epidermal T lymphocyte chemotactic factors rather than a single factor was shown to specifically attract CD4+, CD45R0+ T lymphocytes in contrast to fMLP, IL-8, C5a and LTB4, which induced equal chemotaxis for both CD4+ and CD8+ T lymphocytes. A newly described inhibitory cytokine IL-10 selectively attracted the CD8+ subpopulation of T lymphocytes, and it is suggested that IL-10 could be an important factor in the downregulation of an inflammatory response. The recently discovered neutrophil chemotactic and activating factor IL-8 has been shown to be chemotactic for T lymphocytes as well. It belongs to a family of 8,000-10,000 KDa peptides of which a monocyte chemotactic and activating factor MCAF is also a member. We showed that mRNA for IL-8 could be expressed in highly purified T lymphocytes only upon stimulation with ionomycin and PHA or PMA and PHA, and to a lesser extent PMA and IL-2.

Topics: CD4-CD8 Ratio; CD4-Positive T-Lymphocytes; Cell Line, Transformed; Chemokine CCL2; Chemokines, C; Chemotactic Factors; Chemotaxis, Leukocyte; Cytokines; Humans; Inflammation; Interleukin-10; Interleukin-8; Lymphocyte Activation; Lymphocytes; Lymphokines; Sialoglycoproteins; Skin Diseases; Skin Neoplasms; T-Lymphocyte Subsets

Cytokines are (glyco)proteins that are synthesized and secreted by various cells, which bind to specific receptors on target cells and which regulate activation, proliferation, and differentiation of immune as well as non-immune cells. Keratinocytes upon injury release interleukin (IL)-1, IL-6, IL-8, colony-stimulating factors, and tumor-necrosis factor, as well as growth and suppressor factors. There is also strong evidence for a network of interacting cytokines, which has been only partially characterized so far, maintaining a proper balance. However, excessive or insufficient production of these mediators may contribute to certain disease states, particularly those with infectious and autoimmune genesis. Therefore the understanding of cytokine interactions may be helpful in elucidating the pathomechanisms of such diseases. Moreover, certain cytokines, as well as their analogues and antagonists, may prove to be of therapeutic value.

Topics: Colony-Stimulating Factors; Cytokines; Epidermis; Growth Substances; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Skin Diseases; Suppressor Factors, Immunologic; Tumor Necrosis Factor-alpha

To evaluate the effect of oral Escherichia coli (E. coli) Nissle application on the outcome of intestinal-borne dermatoses.. In a randomized, controlled, non-blinded prospective clinical trial 82 patients with intestinal-borne facial dermatoses characterized by an erythematous papular-pustular rash were screened. At the initiation visit 37 patients entered the experimental arm and 20 patients constituted the control arm. All 57 patients were treated with a vegetarian diet and conventional topical therapy of the dermatoses with ointments containing tetracycline, steroids and retinoids. In the experimental arm patients received a one month therapy with oral E. coli Nissle at a maintenance dose of 2 capsules daily. The experimental group was compared to a non-treatment group only receiving the diet and topical therapy. The primary outcome parameter was improvement of the dermatoses, secondary parameters included life quality and adverse events. In addition the immunological reaction profile (IgA, interleucin-8 and interferon-α) was determined. Furthermore the changes of stool consistency and the microbiota composition over the time of intervention were recorded.. Eighty-nine percent of the patients with acne, papular-pustular rosacea and seborrhoic dermatitis responded to E. coli Nissle therapy with significant amelioration or complete recovery in contrast to 56% in the control arm (P < 0.01). Accordingly, in the E. coli Nissle treated patients life quality improved significantly (P < 0.01), and adverse events were not recorded. The clinical improvement was associated with a significant increase of IgA levels to normal values in serum as well as suppression of the proinflammatory cytokine IL-8 (P < 0.01 for both parameters). In the E. coli Nissle treated group a shift towards a protective microbiota with predominance of bifidobacteria and lactobacteria (> 10(7) CFU/g stool) was observed in 79% and 63% of the patients, respectively (P < 0.01), compared to no change in the control group without E. coli Nissle. Moreover, the detection rate of a pathogenic flora dropped from 73% to 14 % of the patients in the experimental arm (P < 0.01) with no significant change in the control arm (accounting 80% before and 70% after the observation period, P > 0.05). Accordingly, stool consistency, color and smell normalized in the E. coli Nissle treated patients.. E. coli Nissle protects the mucus barrier by overgrowth of a favorable gut microbiota with less immunoreactive potential which finally leads to clinical improvement of intestinal borne dermatoses.

Topics: Administration, Cutaneous; Administration, Oral; Adult; Biological Therapy; Capsules; Diet, Vegetarian; Escherichia coli; Female; Gastrointestinal Microbiome; Humans; Immunoglobulin A; Interferon-alpha; Interleukin-8; Intestinal Diseases; Intestines; Male; Probiotics; Prospective Studies; Quality of Life; Signal Transduction; Skin Diseases; Treatment Outcome

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

In this report, we provide a case of self-limiting canine acute febrile sterile neutrophilic dermatosis in which the clinical signs featured typical target skin lesions with strong upregulation of T-helper 1 markers and interleukin-8, a potent neutrophil chemoattractant. Further, large case series are needed to characterize canine sterile neutrophilic dermatosis.. Dans cet article, nous présentons un cas de dermatose neutrophilique fébrile stérile aiguë canine spontanément résolutive dans laquelle les signes cliniques comportaient des lésions cutanées cibles typiques avec une forte régulation à la hausse des marqueurs T-helper 1 et de l'interleukine-8, un puissant chimioattractant des neutrophiles. D'autres grandes séries de cas sont nécessaires pour caractériser la dermatose neutrophile stérile canine.. En este artículo exponemos un caso de dermatosis neutrofílica estéril febril aguda canina autolimitante en la que los signos clínicos fueron lesiones cutáneas típicas en forma de diana con una intensa elevación de los marcadores T-helper 1 y de interleucina-8, un potente quimiotáctico de neutrófilos. Se necesitan series de casos más grandes para caracterizar la dermatosis neutrofílica estéril canina.. In diesem Bericht beschreiben wir einen Fall selbst-limitierender fieberhafter neutrophiler Dermatose bei einem Hund, wobei die klinischen Zeichen aus typischen Target Hautläsionen mit starker Hochregulierung von T-Helfer 1 Marker und Interleukin-8, welches ein potentes neutrophiles Chemoattractant darstellt, bestanden. Weitere große Fallstudien sind nötig, um die canine sterile neutrophile Dermatose zu charakterisieren.. 本報告では、T-helper 1マーカーおよび好中球誘引物質であるインターロイキン-8の強い発現を伴う典型的な標的皮膚病変を呈した自己限定性犬急性発熱性無菌性好中球性皮膚症の1例を紹介する。犬の無菌性好中球性皮膚症の特徴を明らかにするためには、さらなる大規模なケースシリーズが必要である。.. 在本报告中，我们提供了一例自限性犬急性发热性无菌性中性粒细胞皮肤病，其临床症状表现为典型的靶样皮肤病变，辅助性 T 细胞1标记物和白细胞介素-8(一种强效中性粒细胞趋化因子)明显上调。需要进一步的大规模病例系列来表征犬无菌性中性粒细胞皮肤病。.. Neste relato, nós apresentamos um caso de dermatose neutrofílica estéril canina aguda febril em que os sinais clínicos típicos foram lesões cutâneas em alvo com forte ativação de marcadores T-helper 1 e interleucina-8, um potente quimioatrativo de neutrófilos. São necessários mais estudos com uma grande série de casos para caracterizar a dermatose neutrofílica estéril canina.

Topics: Animals; Dermatitis; Dog Diseases; Dogs; Interleukin-8; Neutrophils; Skin Diseases; Sweet Syndrome; Up-Regulation

Topics: Erythema; Humans; Interleukin-8; Skin; Skin Diseases; Vasculitis, Leukocytoclastic, Cutaneous

Alterations in serum cytokine levels and profiles have been reported in association with a variety of disease conditions (e.g., allergic, immune-mediated, etc.) in both humans and animals. In comparison to serum cytokine measurements, tear cytokine measurements might be expected to more accurately reflect the inflammatory milieu associated with periocular disease. The purpose of this study was to use a multiplexed assay to compare the cytokine profile of tears in healthy dogs to those with inflammatory skin and periocular disease. We were able to detect IL-2, IL-6, IL-8, and TNF-α in >47 % of tear samples from both healthy canine patients and those with inflammatory dermatologic disease (with or without concurrent periocular involvement). In contrast, IL-7, IL-10 and IFN-γ were rarely detected. Dogs with both dermatologic and periocular disease (but not dermatologic disease alone) had higher levels of IL-8 (P < 0.001, P > 0.05, respectively) relative to healthy dogs. Patients with concurrent dermatologic and periocular disease also demonstrated significantly greater variability in IL-8 concentrations between eyes than did healthy dogs (P < 0.0001). Our findings suggest that tear cytokine analysis may prove to be a useful tool to investigate the role and interactions of the local ocular immune response in patients with inflammatory periocular disease.

Topics: Animals; Cytokines; Dog Diseases; Dogs; Eye; Eye Diseases; Female; Interleukin-2; Interleukin-6; Interleukin-8; Male; Skin Diseases; Tears; Tumor Necrosis Factors

Diabetic foot ulcer (DFU) is a common high-risk complication in patients with diabetes mellitus, but current drugs and therapies in management of this disease cannot meet the urgent clinical needs. In this study, a snail glycosaminoglycan (SGAG) from the cultured China white jade snail was purified and structurally clarified. This snail glycosaminoglycan is a regular sulfated polysaccharide, composed of iduronic acid (IdoA) and N-acetyl-glucosamine (GlcNAc) with the repeating sequence of →4)-α-GlcNAc (1→4)-α-IdoA2S (1→. The biological assays showed that SGAG had no anticoagulant activity for lacking specific heparin pentasaccharide sequence. The pharmacological experiments suggested that SGAG markedly accelerated the healing of full-thickness wounds in diabetic mice skin. Histologic and immunohistochemical analysis revealed that SGAG treatment alleviated the inflammation and dermal edema, and promoted angiogenesis. This is the first report applying the snail glycosaminoglycan to favor diabetic wound healing.

Topics: Acetylglucosamine; Actins; Angiogenesis Inducing Agents; Animals; Anti-Inflammatory Agents; Diabetes Mellitus, Experimental; Edema; Epithelium; Glycosaminoglycans; Heparin; Iduronic Acid; Inflammation; Interleukin-8; Magnetic Resonance Spectroscopy; Male; Mice; Platelet Endothelial Cell Adhesion Molecule-1; Regeneration; Skin; Skin Diseases; Snails; Wound Healing

The aim of the study was to investigate the time-course of serum and wound fluids interleukin (IL)-6 and IL-8 levels in dogs with cutaneous wounds and their relationship with some haematologic parameters. The experimental group comprised of six adult dogs that underwent surgery with wounds (n = 6) on the mid lateral aspect of the right antebrachium; and control group of six, apparently, healthy intact (free from cutaneous wounds) adult dogs, comprising equal number of both sexes. Vital signs evaluated were within normal limits. Samples of blood, serum and wound fluids harvested pre- and at 12 h, 36 h, 60 h, 156 h and 324 h post-injury, were utilised for IL-6 and IL-8 assay and haematology. Peak concentrations of IL-6 in wound fluid (1.33. ± 0.33 ng/mL) and serum (0.82 ± 0.24 ng/mL) of the experimental group at 12 h post-operation were higher (P < 0.01) than the control (0.30 ± 0.05 ng/mL). Concentrations of IL-8 at 12 h and 60 h in wound fluid (0.21 ± 0.05 ng/mL and 0.22 ± 0.11 ng/mL) respectively were lower (P < 0.05) than serum (0.71 ± 0.21 ng/mL and 0.73 ± 0.24 ng/mL) respectively in the experimental group and corresponding values recorded in controls (0.34 ± 0.09 ng/mL and 0.36 ± 0.14 ng/mL). The haematological and biochemical parameters exhibited minimum fluctuations, but values were within normal ranges. Significant correlations were obtained between serum and wound fluid IL-6 (r = 0.827, P < 0.05); wound fluid IL-6 and monocyte count (r = 0.818, P < 0.04); wound fluid IL-6 and haematocrit (r = -0.894, P < 0.05). There was a positive correlation between serum IL-8 and serum IL-6 (r = 0.622, P > 0.05) and serum IL-8 and wound fluid IL-8 (r = 0.718, P > 0.05) in the experimental group. In conclusion, IL-6 and IL-8 exerted modulated inflammatory processes following cutaneous wounds in dogs. Further studies are required to investigate the expression patterns of IL-6 and IL-8 in cutaneous wounds in order to improve the quality of management of cutaneous wounds.

Topics: Animals; Dogs; Female; Hematology; Interleukin-6; Interleukin-8; Male; Skin Diseases; Tumor Necrosis Factor-alpha; Wound Healing; Wounds and Injuries

Rhus coriaria L. (R. coriaria) is a medicinal herb native to the middle east and Mediterranean region and well-known as "sumac" or "sicilian sumac". This herb has a wide range of traditional applications, covering its topical use to treat skin burns or eczemas and to promote wound healing.. The present research aims to investigate the potential anti-inflammatory activity of Rhus coriaria L. fruit extracts in human keratinocytes (HaCaT cells), evaluating extracts prepared using different techniques.. Water (WRC), ethanol-water (EWRC) and two types of ethanol extracts (mERC and ERC) were prepared. The HaCaT cells were challenged by TNF-α (10 ng/mL) and IL-8, ICAM-1, VEGF, and MMP-9 release, as well as NF-κB translocation, were measured by ELISA assays. The most active extracts were chemically profiled through HPLC-UV-DAD analysis.. Our results suggest the potential positive effect of R. coriaria fruit extracts, mostly mERC, as preventive agent in the treatment of keratinocyte inflammation through their inhibitory effect on the production of skin pro-inflammatory mediators.

Topics: Anti-Inflammatory Agents; Cell Line; Fruit; Humans; Intercellular Adhesion Molecule-1; Interleukin-8; Keratinocytes; Matrix Metalloproteinase 9; NF-kappa B; Plant Extracts; Rhus; Skin Diseases; Vascular Endothelial Growth Factor A

Some of the most common infectious diseases are caused by bacteria that naturally colonise humans asymptomatically. Combating these opportunistic pathogens requires an understanding of the traits that differentiate infecting strains from harmless relatives. Staphylococcus epidermidis is carried asymptomatically on the skin and mucous membranes of virtually all humans but is a major cause of nosocomial infection associated with invasive procedures. Here we address the underlying evolutionary mechanisms of opportunistic pathogenicity by combining pangenome-wide association studies and laboratory microbiology to compare S. epidermidis from bloodstream and wound infections and asymptomatic carriage. We identify 61 genes containing infection-associated genetic elements (k-mers) that correlate with in vitro variation in known pathogenicity traits (biofilm formation, cell toxicity, interleukin-8 production, methicillin resistance). Horizontal gene transfer spreads these elements, allowing divergent clones to cause infection. Finally, Random Forest model prediction of disease status (carriage vs. infection) identifies pathogenicity elements in 415 S. epidermidis isolates with 80% accuracy, demonstrating the potential for identifying risk genotypes pre-operatively.

Topics: Genome-Wide Association Study; Genome, Bacterial; Genotype; Humans; Interleukin-8; Skin Diseases; Staphylococcal Infections; Staphylococcus epidermidis

Chronic inflammatory skin diseases are characterized by controlled proliferation of keratinocytes. Here, activating transcription factor 3 (ATF3) might play a fundamental role. In these inflammatory diseases, proliferation is controlled and only rarely leads to cancer development which can be supported by an inflammatory microenvironment. ATF3 is a dual function protein as it suppresses pro-inflammatory IL-6 and IL-8, but also acts as a pro-oncogenic factor by the suppression of p53. We therefore analyzed ATF3 expression comparing myeloid cells with keratinocytes.. To dissect the bi-modal role of ATF3 we pharmacologically induced ATF3 and analyzed its influence on cytokine expression and secretion in a cell type specific manner.. Since inflammatory skin diseases can be treated systemically with Cyclosporin A or Dimethylfumarate we stimulated myeloid cells and primary human keratinocytes with these drugs and analyzed gene expression by quantitative real-time PCR. Cytokine secretion was measured by ELISA.. In the present study, we could show that ATF3 is induced in PBMCs by DMF and weakly by Ebselen, while CsA is the most prominent inducer of ATF3 in keratinocytes without enhancing HO-1 transcription. Further we could show that induction of stress by LPS treatment elevates IL-1β and IL-6 and weakly ATF3 transcription in PBMCs. While transcription of both cytokines is elevated, LPS treatment mediates IL-6 secretion with only little IL-1β secretion. Treatment with DMF dampens LPS-induced transcription.. Taken together, our results shed light into the different carcinogenic potential of CsA and DMF, which both target ATF3. Collectively our data demonstrate that CsA strongly induces pro-carcinogenic ATF3 in keratinocytes, whereas ATF3 induction by DMF in myeloid cells acts anti-inflammatory.

Topics: Activating Transcription Factor 3; Carcinogenesis; Cell Proliferation; Cyclosporine; Dermatologic Agents; Dimethyl Fumarate; Enzyme-Linked Immunosorbent Assay; Heme Oxygenase-1; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Keratinocytes; Lipopolysaccharides; Myeloid Cells; Primary Cell Culture; Real-Time Polymerase Chain Reaction; RNA, Messenger; Skin; Skin Diseases; Tumor Suppressor Protein p53; Up-Regulation

Tissue kallikreins (KLKs), in particular KLK5, 7 and 14 are the major serine proteases in the skin responsible for skin shedding and activation of inflammatory cell signaling. In the normal skin, their activities are controlled by an endogenous protein protease inhibitor encoded by the SPINK5 gene. Loss-of-function mutations in SPINK5 leads to enhanced skin kallikrein activities and cause the skin disease Netherton Syndrome (NS). We have been developing inhibitors based on the Sunflower Trypsin Inhibitor 1 (SFTI-1) scaffold, a 14 amino acids head-to-tail bicyclic peptide with a disulfide bond. To optimize a previously reported SFTI-1 analogue (I10H), we made five analogues with additional substitutions, two of which showed improved inhibition. We then combined those substitutions and discovered a variant (Analogue 6) that displayed dual inhibition of KLK5 (tryptic) and KLK7 (chymotryptic). Analogue 6 attained a tenfold increase in KLK5 inhibition potency with an Isothermal Titration Calorimetry (ITC) Kd of 20nM. Furthermore, it selectively inhibits KLK5 and KLK14 over seven other serine proteases. Its biological function was ascertained by full suppression of KLK5-induced Protease-Activated Receptor 2 (PAR-2) dependent intracellular calcium mobilization and postponement of Interleukin-8 (IL-8) secretion in cell model. Moreover, Analogue 6 permeates through the cornified layer of in vitro organotypic skin equivalent culture and inhibits protease activities therein, providing a potential drug lead for the treatment of NS.

Topics: Cell Line; Helianthus; Humans; Interleukin-8; Netherton Syndrome; Peptides, Cyclic; Proteinase Inhibitory Proteins, Secretory; Receptor, PAR-2; Skin; Skin Diseases; Tissue Kallikreins; Trypsin Inhibitors

The prevalence of impaired cutaneous wound healing is high and treatment is difficult and often ineffective, leading to negative social and economic impacts for our society. Innovative treatments to improve cutaneous wound healing by promoting complete tissue regeneration are therefore urgently needed. Mesenchymal stromal cells (MSCs) have been reported to provide paracrine signals that promote wound healing, but (i) how they exert their effects on target cells is unclear and (ii) a suitable delivery system to supply these MSC-derived secreted factors in a controlled and safe way is unavailable. The present study was designed to provide answers to these questions by using the horse as a translational model. Specifically, we aimed to (i) evaluate the in vitro effects of equine MSC-derived conditioned medium (CM), containing all factors secreted by MSCs, on equine dermal fibroblasts, a cell type critical for successful wound healing, and (ii) explore the potential of microencapsulated equine MSCs to deliver CM to wounded cells in vitro.. MSCs were isolated from the peripheral blood of healthy horses. Equine dermal fibroblasts from the NBL-6 (horse dermal fibroblast cell) line were wounded in vitro, and cell migration and expression levels of genes involved in wound healing were evaluated after treatment with MSC-CM or NBL-6-CM. These assays were repeated by using the CM collected from MSCs encapsulated in core-shell hydrogel microcapsules.. Our salient findings were that equine MSC-derived CM stimulated the migration of equine dermal fibroblasts and increased their expression level of genes that positively contribute to wound healing. In addition, we found that equine MSCs packaged in core-shell hydrogel microcapsules had similar effects on equine dermal fibroblast migration and gene expression, indicating that microencapsulation of MSCs does not interfere with the release of bioactive factors.. Our results demonstrate that the use of CM from MSCs might be a promising new therapy for impaired cutaneous wounds and that encapsulation may be a suitable way to effectively deliver CM to wounded cells in vivo.

Topics: Animals; Cell Line; Cell Movement; Cell Proliferation; Cell- and Tissue-Based Therapy; Chemokine CXCL10; Cobalt; Culture Media, Conditioned; Female; Fibroblasts; Gene Expression; Guided Tissue Regeneration; Horses; Interferon-gamma; Interleukin-8; Matrix Metalloproteinase 1; Matrix Metalloproteinase 13; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mitomycin; Models, Animal; Skin; Skin Diseases; Tumor Necrosis Factor-alpha; Wound Healing

Doxycycline is used to treat infective diseases because of its broadspectrum efficacy. High dose administration (100 or 200 mg/day) is often responsible for development of bacterial resistances and endogenous flora alterations, whereas low doses (20-40 mg/day) do not alter bacteria susceptibility to antibiotics and exert anti-inflammatory activities. In this study, we wanted to assess the efficacy of both low and high doxycycline doses in modulating IL-8, TNF-α, and IL-6 gene expression in HaCaT cells stimulated with LPS. Three experimental settings were used, differing in the timing of doxycycline treatment in respect to the insult induced by LPS: pretreatment, concomitant, and posttreatment. Low doses were more effective than high doses in modulating gene expression of LPS-induced proinflammatory cytokines (IL-8, TNF-α, and IL-6), when added before (pretreatment) or after (posttreatment) LPS stimulation. This effect was not appreciated when LPS and doxycycline were simultaneously added to cell cultures: in this case high doses were more effective. In conclusion, our in vitro study suggests that low doxycycline doses could be safely used in chronic or acute skin diseases in which the inflammatory process, either constantly in progress or periodically recurring, has to be prevented or controlled.

Topics: Anti-Inflammatory Agents; Cell Line; Doxycycline; Gene Expression; Humans; Interleukin-6; Interleukin-8; Lipopolysaccharides; Skin Diseases; Tumor Necrosis Factor-alpha

Epidermal growth factor receptor inhibitor (EGFRI) induced skin toxicity has a prognostic value suggesting skin toxicity can be a useful surrogate marker for successful epidermal growth factor receptor (EGFR) inhibition, improved response and survival. But the pathophysiology of EGFRI induced skin toxicity remains elusive. However the involvement of immunological mechanisms has been speculated. This study investigates the possible underlying mechanism of EGFR inhibition associated cytokine production in keratinocytes as well as in patients after treatment with epidermal growth factor receptor inhibitors (EGFRIs).. Normal human epidermal keratinocytes (NHEK) were incubated for 2 and 24h with different concentrations of EGFRI (erlotinib) for Western blot analysis and cytokine expression analysis, respectively. Inhibition of EGFR, extracellular-signal-regulated kinase 1/2 (Erk 1/2) and c-Jun was examined by Western blot analysis. Cytokine concentrations were measured by enzyme-linked immunosorbent assay (ELISA) in the NHEK cell supernatant and also in the serum of 186 cancer patients who are followed up for EGFRI induced skin rash.. A significant inhibitory effect of EGFRI was seen on EGFR (Y845), Erk 1/2 and c-Jun in a dose dependent manner. Further downstream, increased CC-chemokine ligand 2 (CCL2), CC-chemokine ligand 5 (CCL5) and decreased interleukin-8 (IL-8) or CXCL8 expression was observed in keratinocytes. In EGFRI treated patients, low levels of serum CXCL8 corresponding to stronger EGFR inhibition were associated with a higher grade of skin toxicity (p=0.0016) and a prolonged overall survival (p=0.018).. The results obtained in this study indicate that EGFRI can regulate cytokines by modulating EGFR signalling pathway in keratinocytes. Moreover, serum levels of CXCL8 in EGFRI treated patients may be important for individual EGFRI induced skin toxicity and patient's survival.

Topics: Adult; Aged; Aged, 80 and over; Cells, Cultured; Cytokines; Disease-Free Survival; ErbB Receptors; Female; Humans; Interleukin-8; Keratinocytes; Male; Middle Aged; Neoplasms; Prospective Studies; Protein Kinase Inhibitors; Signal Transduction; Skin Diseases

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

Epidermal growth factor receptor (EGFR) inhibitors are widely used in the treatment of cancer. EGFR-targeted treatment is known to be associated with a high incidence of dermatological adverse reactions, including papulopustular rash, which can be dose-limiting and may affect compliance to treatment. Currently, the pathways involved in EGFR inhibitor-induced rash are poorly understood and few treatment options for this adverse event are available. Here, we developed a model for induction of papulopustular rash in healthy human volunteers by subcutaneous injection of the anti-EGFR monoclonal antibody zalutumumab. The injection sites and surrounding skin were evaluated by a dermatologist for the presence or absence of papulopustular rash and skin biopsies were taken to confirm the macroscopical findings by immunohistochemistry. Locally injected zalutumumab induced a papulopustular rash, characterized by acute follicular neutrophil-rich hair follicle inflammation, and thus mimicked adverse events induced by systemic administration of EGFR inhibitors. In this model, we tested the hypothesis that neutrophils, attracted by IL-8, play a central role in the observed rash. Indeed, concomitant local repeat dose treatment with HuMab-10F8, a neutralizing human antibody against IL-8, reduced the rash. Inhibition of IL-8 can therefore ameliorate dermatological adverse events induced by treatment with EGFR inhibitors.

Topics: Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; ErbB Receptors; Humans; Injections, Subcutaneous; Interleukin-8; Neutralization Tests; Skin Diseases

The purpose of this study was to investigate the effects of size and phase composition on human exposure to airborne titanium dioxide (TiO(2)) nanoparticles (NPs) at workplaces. We reanalyzed published data of particle size distribution of airborne TiO(2) NPs during manufacturing activities and linked a physiologically based lung model to estimate size- and phase-specific TiO(2) NP burdens in target lung cells. We also adopted a cell model to simulate the exposure time-dependent size/phase-specific cell uptake of TiO(2) NPs in human dermal and lung cells. Combining laboratory, field, and modeling results, we proposed two major findings: (i) the estimated median effective anatase TiO(2) NP concentration (EC50) for cytotoxicity response on human dermal fibroblasts was estimated to be 24.84 (95% CI: 7.3-70.2) nmolmL(-1) and EC50 estimate for inflammatory response on human lung epithelial cells was 5414 (95% CI: 3370-7479) nmolmL(-1) and (ii) packers and surface treatment workers at the TiO(2) NP production workplaces are unlikely to pose substantial risk on lung inflammatory response. Nevertheless, our findings point out that TiO(2) NP production workers have significant risk on cytotoxicity response at relatively high airborne anatase TiO(2) NP concentrations at size range 10-30nm.

Topics: Algorithms; Cells, Cultured; Cytosol; Dose-Response Relationship, Drug; Endosomes; Humans; Interleukin-8; Lung; Lysosomes; Nanoparticles; Occupational Exposure; Particle Size; Pulmonary Alveoli; Risk Assessment; Skin Absorption; Skin Diseases; Titanium

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

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

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

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

Alpha-melanocyte-stimulating hormone (alpha-MSH) is a neuropeptide predominantly produced by the pituitary gland, but it is also generated by many extra-pituitary cells including keratinocytes of the skin. This neuropeptide has anti-inflammatory and antimicrobial effects and probably contributes in innate immunity. Staphylococcus aureus is the aetiological agent of a wide range of infections in humans. Colonization of human skin by S. aureus is a characteristic feature of several skin diseases and is often followed by tissue invasion and severe cell damage. The aim of our study was to detect a possible role of alpha-MSH during the early infection stages in the adhesion and penetration of keratinocytes before cell damage. Our data demonstrated that alpha-MSH precociously down-regulates the production of integrins such as beta1 and heat shock surface protein 70, essential molecules for the entry of S. aureus. Moreover, in our experimental model, alpha-MSH induces the down-regulation of the pro-inflammatory cytokine expression and of the adhesion molecules in keratinocytes activated by S. aureus. Our data suggest that alpha-MSH plays a protective role in the skin by reducing infection and the inflammatory process.

Topics: alpha-MSH; Blotting, Western; Cell Line; Cell Line, Tumor; Cell Survival; Cytokines; Down-Regulation; Enzyme-Linked Immunosorbent Assay; HSP70 Heat-Shock Proteins; Humans; Inflammation; Integrins; Intercellular Adhesion Molecule-1; Interleukin-8; NF-kappa B; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Skin Diseases; Staphylococcus aureus; Tumor Necrosis Factor-alpha

Topics: Candida albicans; Candidiasis; Cells, Cultured; Coculture Techniques; Gene Expression Regulation; Humans; Interleukin-8; Keratinocytes; Kinetics; RNA, Messenger; Skin Diseases; Time Factors

We have developed a simple noninvasive method to assess inflammatory changes in human skin, even in the absence of visible clinical irritation. Our approach is based on a simple tape (Sebutape) adsorption method to recover molecular mediators of skin inflammation (e.g., cytokines). This procedure has been used to investigate baseline cytokine levels on skin, to assess normal skin condition and to evaluate changes due to chemical insult, existing dermatitis, or sun exposure.. In clinical studies, Sebutape was applied to normal appearing uncompromised skin, as well as to compromised (diaper or heat rash), chemically treated (sodium laurel sulfate), or sun-exposed skin. Sebutape was applied to the skin for a 1 min collection interval. Tapes were extracted in saline using a 10 min sonication, and the extracts were analyzed for human interleukin-1alpha (IL-1alpha), IL-1 receptor antagonist (IL-1RA) and IL-8 using commercial immunoassay test kits. The cytokine levels recovered from each tape extract were normalized to total protein (TP) levels. In infant product use tests, the severity of skin irritation (diaper and heat rash or erythema) was also assessed using a visual grading scale.. The method itself caused minimal, if any, skin damage. Additionally, Sebutape was shown to quantitatively adsorb detectable levels of cytokine from normal-appearing (control) or compromised (e.g., rashed or chemically treated) skin. In infant studies, significant increases in IL-1alpha levels were found in skin exhibiting diaper rash, heat rash and erythema compared with normal appearing control skin sites. When these results were normalized to total protein levels recovered from each tape, the significance was maintained. A positive correlation (r2=0.82) existed between IL-1RA levels and diaper rash severity. Significant increases in IL-8 levels were recovered from diaper rash versus control skin sites. There were differences in baseline cytokine levels in normal skin related to body site and sun exposure. The IL-1 RA/IL-1alpha ratios for sun-exposed skin of the face and lower leg were significantly (P<0.05) higher (3-6-fold) than those for skin sites that typically receive minimal sun exposure (i.e., underarm, upper leg and upper back). There was a significant increase in IL-1alpha and a directional increase in IL-8 levels in adult skin sites treated with the irritant, sodium lauryl sulfate, even in the absence of visible skin irritation (erythema).. Our results demonstrate that this method is a useful noninvasive technique for assessing skin inflammatory events. In addition, the method is simple and easily applied in a clinical setting, whether on infants or adults.

Topics: Adhesives; Adsorption; Aged; Biomarkers; Cytokines; Dermatitis; Environmental Exposure; Forearm; Humans; Infant; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Interleukin-8; Sialoglycoproteins; Skin; Skin Diseases; Sodium Dodecyl Sulfate; Sunlight; Surface-Active Agents; Time Factors

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 participation of apoptotic cell death of neutrophils in the development of skin lesions of patients with anaphylactoid purpura was examined by the in situ specific labeling of fragmented DNA. In the early stage of the skin lesions, there were few positively stained nuclei in infiltrating cells. The number of positive cells increased markedly in the fully developed stage of the lesions. A number of neutrophils were stained positively. Finally, a few fragmented nuclei were still positive in the late stage of the lesions. It was therefore suggested that fragmentation of neutrophils in the skin lesions from the patients might be due to apoptosis. Inducible nitric oxide synthase and nitrotyrosine were detected in infiltrates, and interleukin-8 was also detected in vascular endothelial cells in those skin lesions. The roles of nitric oxide and interleukin-8 in the apoptosis of neutrophils are discussed.

Topics: Adolescent; Adult; Aged; Anaphylaxis; Apoptosis; Cell Nucleus; Child; Child, Preschool; DNA Fragmentation; Endothelium, Vascular; Female; Humans; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Nitric Oxide Synthase; Purpura; Skin Diseases; Tyrosine

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

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

Interleukin-8 (IL-8) is a cytokine with potent neutrophil chemotactic and activating properties and is active in inflammatory conditions in man. It has been identified in human inflammatory skin conditions where it is likely to be responsible for both neutrophil recruitment from the circulation and possibly T-lymphocyte chemoattraction. Studies in animals also suggest that IL-8 may augment skin oedema.. To study the effects of intradermally administered IL-8 in humans on tissue oedema and cellular recruitment in atopic and non-atopic volunteers.. Interleukin-8 (1.2 x 10(-7) M) in the presence and absence of histamine was administered by intradermal injection. Wheal and erythema area were measured at regular intervals and 3 h following challenge punch biopsies were taken for immunocytochemistry. Cellular infiltrate was measured by immunocytochemical identification of neutrophils, eosinophils and T-lymphocytes in glycol-methacrylate-embedded sections.. In the presence of histamine, IL-8 provoked a significantly greater wheal area when compared to that produced by histamine alone (P < 0.001). In the presence of histamine, IL-8 produced a significantly greater neutrophil infiltrate (P < 0.05), however, neither lymphocyte or eosinophil infiltration was found to be increased with IL-8 challenge. There was no difference observed between atopic and non-atopic subjects, nor were any effects of IL-8 demonstrated in the absence of histamine.. This study demonstrates that in human skin, IL-8 induces increased microvascular permeability and neutrophil infiltration, but not eosinophil or T-lymphocyte chemoattraction.

Topics: Adolescent; Adult; Allergens; Capillary Permeability; Chemotaxis, Leukocyte; Edema; Eosinophils; Histamine; Humans; Hypersensitivity; Immunohistochemistry; Injections, Intradermal; Interleukin-8; Male; Middle Aged; Neutrophil Activation; Neutrophils; Recombinant Proteins; Skin Diseases; T-Lymphocytes

IL-8 is a member of the chemokine alpha subfamily that activates and is chemotactic for neutrophils. In these studies, we have synthesized and characterized a hexapeptide inhibitor of IL-8. This peptide, with an acetylated amino terminus and an amidated carboxyl terminus (Ac-RRWWCR-NH2), inhibited the specific binding of 125I-IL-8 to neutrophils. The inhibition was biphasic and apparent Ki was estimated to be approximately 2.7 microM and 13 microM for two different IL-8 binding sites. The peptide inhibited neutrophil chemotaxis, beta-glucuronidase release from neutrophils, and rabbit skin edema induced by IL-8 with an EC50 of 90 microM, 0.8 microM, respectively. Ac-RRWWCR-NH2 also suppressed the binding of macrophage inflammatory protein (MIP) 2 beta to neutrophils. However, it did not inhibit the binding of MIP-1 alpha, C5a, or leukotriene B4 to neutrophils, chemotaxis induced by FMLP, or beta-glucuronidase release induced by FMLP, C5a, or leukotriene B4. Additional peptides were analyzed to identify a better inhibitor. Inhibition of binding by Ac-rrwwcrc-NH2 synthesized with all D-amino acids was almost four times more potent than Ac-RRWWCR-NH2. Small peptide homologues of the amino-terminal end of IL-8 failed to inhibit IL-8 binding to neutrophils. These studies have identified several peptides that significantly inhibit IL-8 function. Because IL-8 seems to be an important inflammatory mediator of several human illnesses, these peptides may have pharmacologic potential.

Topics: Amino Acid Sequence; Cell Line; Chemotaxis, Leukocyte; Edema; Humans; Interleukin-8; Molecular Sequence Data; Neutrophil Activation; Neutrophils; Oligopeptides; Receptors, Interleukin; Receptors, Interleukin-8A; Skin Diseases

In this study we report on functional characteristics of pustule as well as blood polymorphonuclear neutrophils (PMN) in a patient suffering from relapsing bullous staphyloderma. Large numbers of viable PMN from newly formed pustules as well as from the peripheral blood were investigated. During the course of disease chemotactic migration, enzyme degranulation, superoxide-anion generation and leukotriene B4 production were determined simultaneously. The results revealed C5a- and NAP-1/IL-8-specific dysfunction of pustule PMN as compared with blood PMN. In contrast, FMLP-elicited functional activities of pustule PMN were only slightly affected. Our findings provide evidence that in inflamed tissue invading PMN are regulated by in situ generated mediators. C5a produced by staph, aureus-induced activation of the alternative pathway of the complement cascade represents a predominant regulatory factor in situ. Furthermore, the results substantiate previous observations concerning different modulation of C5a and f-met-peptide receptors on human PMN.

Topics: Chemotaxis, Leukocyte; Complement C5a; Glucuronidase; Humans; Interleukin-8; Leukotriene B4; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Skin Diseases; Superoxides

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