Wolfgang Czech

High-dose UVA1 therapy in the treatment of patients with atopic dermatitis

BACKGROUND Besides glucocorticosteroids, there is currently no known effective therapy for patients with acute atopic dermatitis. OBJECTIVE The therapeutic effectiveness of high-dose UVA1 irradiation in the management of patients with acute exacerbation of atopic dermatitis was examined. METHODS Patients in the high-dose UVA1 group (n = 15) were irradiated with 130 joules/cm2 UVA1; the control group (n = 10) was treated with UVA-UVB therapy in a minimal erythema dose-dependent manner (total number of treatments 15). RESULTS High-dose UVA1 irradiation was found to induce a significant clinical improvement of atopic dermatitis (p less than 0.001). In comparison with UVA-UVB therapy, significant differences in favor of high-dose UVA1 were observed (p less than 0.01). High-dose UVA1, but not UVA-UVB treatment, significantly reduced the elevated serum level of eosinophil cationic protein in patients with atopic dermatitis (p less than 0.003). CONCLUSION These studies indicate that high-dose UVA1 irradiation may represent a novel phototherapeutic modality for the treatment of patients with an acute exacerbation of atopic dermatitis.

High-dose UVA1 therapy for atopic dermatitis: Results of a multicenter trial

BACKGROUND The results of an open, single-center study suggested that phototherapy with high doses of UVA1 radiation (UVA1R; 340-400 nm) is effective for acute, severe exacerbations of atopic dermatitis (AD). OBJECTIVE The purpose of this study was to assess the effectiveness of high-dose UVA1 phototherapy for acute, severe AD in a randomized multicenter trial in direct comparison with topical glucocorticoid therapy. METHODS Patients were treated with high-dose UVA1R (10 days, 130 J/cm2/day; n = 20), topically with fluocortolone (10 days, 1 x daily; n = 17), or with UVA-UVB therapy (10 days, 1 x daily, minimal erythema dose-dependent; n = 16). RESULTS With a clinical scoring system, significant differences in favor of high-dose UVA1R and fluocortolone therapy were observed (p < 0.0001), as compared with UVA-UVB therapy. At day 10, high-dose UVA1R was superior to fluocortolone (p < 0.002) therapy. Serum levels of eosinophil cationic protein and the blood eosinophil count were significantly reduced after high-dose UVA1 or fluocortolone, but not UVA-UVB therapy. CONCLUSION This study confirms the therapeutic effectiveness of high-dose UVA1 monotherapy for treatment of severe exacerbations of AD.

Serum eosinophil cationic protein (ECP) is a sensitive measure for disease activity in atopic dermatitis.

Atopic dermatitis (AD) is characterized by alterations in cellular and humoral immunity including elevated serum levels of IgE, IL‐2 receptor (IL‐2R) and eosinophil cationic protein (ECP). In order to evaluate the relevance of these serum parameters as indicators of disease activity, the concentrations of IgE, IL‐2R and ECP were measured in serum samples of patients with an acute exacerbation of AD (n=19) on admission to hospital and every 6 days up to discharge, and compared with those from normal non‐atopic controls (n= 15). The severity of the disease in the AD patients was examined using an established clinical scoring system. On admission, AD patients showed significantly elevated serum levels of IgE, IL‐2R and ECP compared with normal controls (P≤0.0001). Clinical improvement was associated with a decrease of both the clinical score (P≤0.001) and serum ECP levels (P≤0.005). No significant changes in serum IgE and serum IL‐2R were observed. In addition, there was a significant correlation between serum ECP and the clinical score (R=0.67, P≤0.001). These data indicate that serum ECP may be a helpful tool for monitoring disease activity in AD.

Eosinophil cationic protein in sera of patients with atopic dermatitis

Patients with atopic dermatitis frequently show elevated blood eosinophil counts, and eosinophil-derived major basic protein has been demonstrated in the eczematous skin from patients with atopic dermatitis. To evaluate further the role of eosinophils in the pathogenesis of atopic dermatitis, the concentration of eosinophil cationic protein was measured in serum samples of 42 patients with moderate to severe disease. The results were compared with those obtained in 32 patients with psoriasis with (n = 9) or without (n = 23) a history of inhalant allergy, 12 patients with a history of pseudoallergic reactions to acetylsalicylic acid, 14 patients with a history of inhalant allergy, and 31 nonatopic healthy control subjects. Eosinophil cationic protein levels were significantly increased in the serum of patients with atopic dermatitis (p less than or equal to 0.005) and patients with a history of pseudoallergic reactions to acetylsalicylic acid (p less than or equal to 0.01). There was no significant difference between eosinophil cationic protein levels in patients with psoriasis or a history of inhalant allergy and in control subjects. Moreover, eosinophil cationic protein levels did not differ significantly in psoriasis patients with or without inhalant allergy. These studies support the concept of an active participation of eosinophils in atopic dermatitis and point to a possible role for eosinophils in pseudoallergy.

Granulocyte activation in bullous diseases: Release of granular proteins in bullous pemphigoid and pemphigus vulgaris

BACKGROUND Eosinophil and polymorphonuclear granulocytes may be involved in the formation of blisters in bullous dermatoses, particularly bullous pemphigoid. OBJECTIVE Our purpose was to evaluate the role of granulocyte activation in the pathogenesis of pemphigus vulgaris and bullous pemphigoid. METHODS Levels of eosinophil cationic protein (ECP) and neutrophil-derived myeloperoxidase (MPO) in blister fluid and serum and levels of serum IgE were determined in patients with bullous pemphigoid (n = 12), those with pemphigus vulgaris (n = 9) and healthy volunteers (n = 12). RESULTS In blister fluid and serum of patients with bullous pemphigoid, significantly elevated concentrations of ECP, MPO and IgE were detected as compared with controls. In contrast, ECP, MPO, and IgE levels in blister fluid and serum of patients with pemphigus vulgaris did not significantly differ from controls. Moreover, the MPO/ECP ratio in serum of patients with bullous pemphigoid was significantly decreased as compared with controls, whereas the MPO/ECP ratio in pemphigus vulgaris did not differ from controls, indicating a preferential activation of eosinophils in bullous pemphigoid only. In patients with bullous pemphigoid, serum levels of ECP and MPO significantly decreased during immunosuppressive therapy to levels similar to those of controls. CONCLUSION Activated granulocytes, releasing their granular contents such as ECP and MPO, may be of importance for blister formation in bullous pemphigoid and may be useful for monitoring disease activity.

Release of sulfidoleukotrienes in vitro: Its relevance in the diagnosis of pseudoallergy to acetylsalicylic acid

Pseudo-allergic reactions (PAR) are caused by a variety of drugs, of particular interest by acetylsalicylic acid (ASA) and other nonsteroidal antiinflammatory drugs. The clinical symptoms often resemble immediate type hypersensitivity reactions and consist of bronchospasm, urticaria, angioedema and even anaphylactic shock. Antigen specific immune mechanisms, however, are not involved. In general, skin tests are not reliable and the diagnosis of PAR is based mainly on risky provocation tests. Therefore, the purpose of this study was to establish procedures for in vitro diagnosis of PAR to ASA. A controlled study was performed including patients with PAR to ASA based on history and positive oral provocation test and non-atopic as well as atopic controls. In this in vitro study the production of sulfidoleukotrienes (sLT) by isolated leukocytes was measured by cellular allergen stimulation test (CAST), which is based on detection of LTC4, LTD4 and LTE4 by a monoclonal antibody. Accordingly, the direct effect of ASA as well as the modulatory effect of ASA on C5a-induced production of sLT in leukocytes in vitro was investigated. In patients with PAR to ASA, C5a-induced generation of sLT was significantly increased as compared to normal controls. In contrast, there was no difference in the spontaneous release of sLT in vitro in patients and controls. Preincubation of leukocytes with ASA did not exert a significant modulatory effect on the spontaneous or the C5a-induced production of sLT in patients and controls. In summary, the present study provides a novel in vitro test system for the diagnosis of PAR to ASA by measurement of sLT release in leukocytes. Moreover, it is attempting to speculate, that C5a induced production of sLT might be a crucial step in the pathogenesis of PAR to ASA in vivo.

Soluble E‐selectin in sera of patients with atopic dermatitis and psoriasis—correlation with disease activity

Summary E‐selectin endothelial leucocyte adhesion molecule‐1 is expressed on endothelial cells in distinct inflammatory skin diseases. E‐selectin mediates the adhesion between activated endothelium and different inflammatory cells. To evaluate soluble E‐selectin as a marker of disease activity in patients with atopic dermatitis and psoriasis, the concentration of soluble E‐selectin, determined by ELISA, was studied in sera of patients before and after treatment and compared with normal non‐atopic controls. The disease severity was established using clinical scoring systems. Levels of soluble E‐selectin were significantly elevated in sera of patients with atopic dermatitis and psoriasis (as compared with controls). Clinical improvement, after treatment, in patients with atopic dermatitis, but not in psoriasis, was associated with a significant decrease in serum levels of soluble E‐selectin. There was a significant correlation of soluble E‐selectin and disease activity in patients with atopic dermatitis. These data indicate that soluble E‐selectin is another parameter to evaluate the inflammatory response in atopic dermatitis and psoriasis. Determination of soluble E‐selectin may be a useful measure of disease activity in atopic dermatitis.

Modulation of intercellular adhesion molecule 1 (ICAM-1) expression on the human mast-cell line (HMC)-1 by inflammatory mediators

The effect of inflammatory mediators on the expression of several surface adhesion molecules on the human mast‐cell line (HMC)‐1 was studied. By flow cytometry, it could be shown that among several surface adhesion molecules (ICAM‐UCDS4, VLA‐4/CD49d, Mac‐UCD11b, LFA‐1/CD11a, LFA‐2/CD2, LFA‐3/CDS8, VCAM‐1), only the constitutively expressed immunoglobulin family member intercellular adhesion molecule‐1 (ICAM‐1) is modulated by proinflammatory cytokines on HMC‐1 mast cells. Stimulation with tumor necrosis factor‐a (TNF‐α) and interferon‐γ (IFN‐γ) resulted, in addition to interleukin‐(lL‐)4, in selective upregulation of ICAM‐1 expression. Costimulation of either IL‐4 or IFN‐γ with TNF‐α further increased the ICAM‐1 expression as compared to the stimuli alone. In contrast, stem‐cell factor (SCF), granulocyte/macrophage colonystimulating factor (GM‐CSF), IL‐10, IL‐8, monocyte chemotactic and activating factor (MCAF), and the complement split product C5a failed to modulate the expression of any adhesion molecule examined. The levels of cytoplasmic free calcium in HMC‐1 mast cells were not altered by cross‐linking surface ICAM‐1, suggesting linkage of other intracellular signaling pathways. This cytokine‐induced upregulation of ICAM‐1 expression might reveal a putative regulatory mechanism of mast‐cell interaction with effector cells bearing the counterparts of ICAM‐1 (CD54), the molecules Mac‐1 (CD11b/CD18) and leukosialin (CD43), and the principal ligand LFA‐1 (CD11a/CD18).

TNFα‐induced activation of eosinophil oxidative metabolism and morphology ‐ Comparison with IL‐5*

Abstract Human dermal mast cells are capable of releasing cytokines, particularly preformed TNFα, upon appropriate stimulation. Masl cell activation in vivo was shown to be associated with an influx and activation of inflammatory cells, initially PMN “polymorphonuclear neutrophilic granulocytes” then eosinophils. In order to learn more about the mechanisms by which TNFα is capable of activating eosinophils, in the present study the effect of TNFα on morphology and function of highly purified normal eosinophils “≥95%” was examined. As estimated by transmission and scanning electron microscopy, TNFα‐stimulated eosinophils appeared to be strictly adherent and flattened exhibiting a characteristic “hemispheric” shape. TNFα induced a dose‐dependent, long‐lasting production of reactive oxygen species as measured by lucigenin‐dependent chemiluminescence (CL). even at a concentration of 0.001 U/ml. The maximal response upon stimulation with TNFα, however, was significantly lower than optimal effects induced by IL‐5. TNFα‐induced responses were completely inhibited by cytochalasin B and staurosporin, and partially blocked by pertussis toxin. Separation of eosinophils by discontinuous density gradients revealed the existence of at least two hypodense eosinophil populations with a distinct susceptibility to stimulation with TNFα. Based on functional assay systems, in contrast to a significant extracellular, only a small intracellular H2O2: production was detected. Accordingly, H2O2 production, detected by an ultrastructural technique, was observed only on the outer surface of the plasma membrane in the contact zones in between adjacent cells. Extracellular as well as intracellular production of H2O2; was completely inhibited by cytochalasin B. TNFα‐induced activation of eosinophils is most probably mediated by binding to the 55 kD and the 75 kD TNF‐receptor since both receptor molecules could be detected by FACS analysis and immune electron microscopy using receptor‐specific antibodies. However, in contrast to its effect on eosinophil oxidative response, TNFα did not induce the release of significant concentrations of eosinophil cationic protein or eosinophil peroxidase in supernatants of cytokine‐stimulated eosinophils, as detected by functional as well as immunological assay systems. These results clearly indicate that TNFα represents a potent eosinophil‐activating cytokine which may be of relevance in the allergic inflammatory response.

Downregulation of platelet-activating factor responsiveness during maturation of human dendritic cells†

Dendritic cells (DCs) are specialized antigen‐presenting cells characterized by their ability to migrate into target sites, process antigens, and activate naive T‐cells. Biological activities of platelet‐activating factor (PAF) and the cytokine macrophage inflammatory protein‐3β (MIP‐3β) as well as the mRNA expression of their receptors were characterized in human DCs during lipopolysaccharide (LPS)‐promoted maturation. Platelet‐activating factor induced calcium transients, migration‐associated actin polymerization response, and chemotaxis in immature human dendritic cells differentiated in vitro from monocytes with interleukin‐4 and granulocyte macrophage colony stimulating factor. In addition, RT‐PCR experiments indicated mRNA expression of the PAF receptor in these immature DCs. Cell studies and mRNA analyses further revealed that immature DCs neither respond to MIP‐3β nor express its specific receptor, CCR7. Induction of cell differentiation by LPS led to the loss of the mRNA expression of the PAF receptor, accompanied by decreasing intracellular calcium release, actin polymerization, and migration after stimulation with PAF. In contrast, LPS treatment induced increasing responsiveness toward MIP‐3β and mRNA expression of CCR7. Comparable data regarding mRNA expression of PAF receptor and PAF responsiveness were also obtained with another maturation protocol using TNFα instead of LPS. The direct comparison between the two different protocols showed a slower decrease of PAF responsiveness induced by TNFα than by LPS. These results show the loss of PAF responsiveness associated with downregulation of PAF receptor mRNA expression during LPS‐ and TNFα‐induced maturation in human DCs. Therefore, these findings point to a functional relevance of PAF in recruiting immature DCs, whereas MIP‐3β might regulate the migration of DCs at a later stage of maturation. J. Cell. Physiol. 185:394–400, 2000.