Allergic sensitivity in different groups of patients with atopic dermatitis

69 patients with atopic dermatitis, aged 16-42 years, were examined. The patients were divided into two groups: group 1 consisted of patients with atopic dermatitis and bronchial asthma and/or allergic rhinitis (25 patients), group 2 consisted of patients without the respiratory syndrome (44 patients). Scarification skin tests made it possible to find out the essential difference in the sensitivity of the examinees in these two groups. In group 1 the prevalence of sensitization to house-dust mites in 23 patients (92%) with monosensitization in 8 patients (32% of the group) was observed. In group 2 sensitivity of house-dust mites was lower: it was registered in 16 patients (36%) with monosensitization in 2 patients (4% of the group). The presence of cross-sensitivity between mite allergens and Candida albicans was established. In accordance with the results of scarification skin tests, treatment included the use of antihistamine preparations, antipruritic remedies and hormonal ointments as well as the elimination of sensitizing allergens. Improvement was registered in 21 patients.     

Increased acetylcholine levels in skin biopsies of patients with atopic dermatitis

Recent experimental evidence indicates that non-neuronal acetylcholine is involved in the regulation of basic cell functions. Here we investigated the cholinergic system in the skin of healthy volunteers and patients with atopic dermatitis (AD). The synthesizing enzyme, choline-acetyltransferase (ChAT), was studied by anti-ChAT immunohistochemistry and enzyme assay. Skin biopsies taken from healthy volunteers and from AD patients were separated into the 2 mm superfical (epidermis and upper dermis) and 3 mm underlying portion (deeper dermis and subcutis). ChAT enzyme activity was detected in homogenized skin and subcutaneous fat (about 13 nmol/mg protein/h). ChAT immunoreactivity was expressed in keratinocytes, hair papilla, sebaceous and eccrine sweat glands, endothelial cells and mast cells. In healthy volunteers the superficial and underlying portion of skin biopsies contained 130 +/- 30 and 550 +/- 170 pmol/g acetylcholine (n = 12), respectively. In AD patients (n = 7) acetylcholine was increased 14-fold in the superficial and 3-fold in the underlying biopsy portion. The present study demonstrates the widespread expression of ChAT protein in the vast majority of human skin cells. Tissue levels of acetylcholine are greatly (14-fold) enhanced in the superficial 2 mm skin of AD patients.     

Sphingosylphosphorylcholine is upregulated in the stratum corneum of patients with atopic dermatitis

To clarify the functional relevance of sphingomyelin (SM) deacylase to the ceramide deficiency seen in atopic dermatitis (AD), we developed a new highly sensitive method and measured the metabolic intermediate sphingosylphosphorylcholine (SPC) that accumulates in the stratum corneum. SPC in intercellular lipids extracted from stratum corneum was reacted with [(14)C]acetic anhydride to yield [(14)C-C(2)]SM, which was then analyzed by TLC. In both the lesional and non-lesional stratum corneum obtained from patients with AD, there was a significant increase in the content of SPC over that of healthy control subjects. There was a reciprocal relationship between increases in SPC and decreases in ceramide levels of stratum corneum obtained from healthy controls, and from lesional and non-lesional skin from patients with AD. Comparison with other sphingolipids present in the stratum corneum demonstrated that there is a significant positive correlation between SPC and glucosylsphingosine, another lysosphingolipid derived from glucosylceramide by another novel epidermal enzyme, termed glucosylceramide deacylase. In contrast, there was no correlation between SPC and sphingosine, a degradative product generated from ceramide by ceramidase. These findings strongly suggest the physiological relevance of SM deacylase function in vivo to the ceramide deficiency found in the skin of patients with AD.     

Clinical and immunological changes in atopic dermatitis patients treated with purified staphylococcal toxoid

32 patients with atopic dermatitis (AD), complicated by pyoderma, were treated with purified staphylococcal toxoid (PST). Changes in clinical and laboratory characteristics in the course of treatment were evaluated. PST was shown to produce a satisfactory therapeutic effect, arresting the symptoms of local staphylococcal infection. An increase in the levels of alpha- and gamma-interferons and decreased content of CD25+ lymphocytes were found. Thus prospects appear for using this preparation as an interferon inductor, as well a for the immunotherapy of AD patients sensitized to Staphylococcus aureus.     

No effect of mobile phone-like RF exposure on patients with atopic dermatitis

This study investigates the effect of exposure to a mobile phone-like radiofrequency (RF) electromagnetic field on people with atopic dermatitis (AD). Fifteen subjects with AD were recruited and matched with 15 controls without AD. The subjects were exposed for 30 min to an RF field at 1 W/kg via an indoor base station antenna attached to a 900 MHz GSM mobile phone. Blood samples for ELISA analysis of the concentration of substance P (SP), tumor necrosis factor receptor 1 (TNF R1), and brain derived neurotrophic factor (BDNF) in serum were drawn before and after the provocation (exposure/sham). Baseline heart rate and heart rate variability, local blood flow, and electrodermal activity were also recorded. No significant differences between the subject groups were found for baseline neurophysiological data. The cases displayed a serum concentration of TNF R1 significantly higher than the control subjects and a significantly lower serum concentration of BDNF in the baseline condition. For SP there was no difference between groups. However, no effects related to RF exposure condition were encountered for any of the measured substances. As to symptoms, a possible correlation with exposure could not be evaluated, due to too few symptom reports. The result of the study does not support the hypothesis of an effect of mobile phone-like RF exposure on serum levels of SP, TNF R1, and BDNF in persons with AD.     

Atopy patch test with Dermatophagoides pteronyssinus (Dp 1) in atopic dermatitis patients

The aim of this study was to evaluate the role of Dermatophagoides pteronyssinus (Dp) in atopic dermatitis patients, using atopy patch test (APT) with Dp (extract 1). Twenty patients (males (m) = 9, females (f) = 11, mean age = 46.0 years, range = 19-78 years) with atopic dermatitis were involved in this study. The control group consisted of seventeen healthy subjects (m = 7, f = 10, mean age = 48.3, range = 24-64 years), with no personal or family history and no signs of atopy. Total IgE, specific IgE and a skin prick test were done for all subjects involved in this study. The atopy patch tests were performed with Dp (extract 1) in: 3,000, 10,000, 20,000 and 30,000 biological units per ml (BU/ml) concentrations using glycerol as medium. The total IgE was significantly higher in atopic dermatitis (AD) patients than in a control group with (p < 0.05). After the tests six of twenty patients (30%) had positive APT results in the last two concentrations (20,000 and 30,000 BU/ml). However, all the results were positive after 48 h (and 72 hours), while no positive results were recorded in the control subjects. According to our study, APT with Dp 1 in 20,000 BU/ml and reading time 48 h and 72 hours is to be recommended. The results suggest that APT may detect the trigger factor (Dp) in AD patients.     

Cholesterol selectively enhances in vitro latex-specific IgE production in atopic dermatitis patients with latex allergy

Effect of cholesterol on in vitro latex-specific IgE production by mononuclear cells from atopic dermatitis patients with latex allergy. Cholesterol enhanced latex-specific IgE production in a dose-dependent fashion, and maximal enhancement was achieved at 1 microg/ml. In contrast, cholesterol had no effect on latex -specific IgA or IgG4 production. Study for cytokine production revealed that cholesterol decreased latex-induced production of IFN-gamma and IL-12, while it increased latex-induced production of IL-4, IL-10 and IL-13. These results indicate that cholesterol skews cytokine pattern toward Th2 type. Collectively, cholesterol may increase allergen-specific IgE production, which may in turn aggravate allergic symptoms.     

Characterization of the skin fungal microbiota in patients with atopic dermatitis and in healthy subjects

Patients with atopic dermatitis (AD) are highly susceptible to viral, bacterial, and fungal skin infections because their skin is dry and this compromises the barrier function of the skin. Therefore, the skin microbiota of patients with AD is believed to be different from that of healthy individuals. In the present study, the skin fungal microbiota of nine patients with mild, moderate, or severe AD and ten healthy subjects were compared using an rRNA clone library. Fungal D1/D2 large subunit analysis of 3647 clones identified 58 species and seven unknown phylotypes in face scale samples from patients with AD and healthy subjects. Malassezia species were predominant, accounting for 63%-86% of the clones identified from each subject. Overall, the non-Malassezia yeast microbiota of the patients was more diverse than that of the healthy individuals. In the AD samples 13.0 ± 3.0 species per case were detected, as compared to 8.0 ± 1.9 species per case in the samples taken from healthy individuals. Notably, Candida albicans, Cryptococcus diffluens, and Cryptococcus liquefaciens were detected in the samples from the patients with AD. Of the filamentous fungal microbiota, Cladosporium spp. and Toxicocladosporium irritans were the predominant species in these patients. Many pathogenic fungi, including Meyerozyma guilliermondii (anamorphic name, Candida guilliermondii), and Trichosporon asahii, and allergenic microorganisms such as Alternaria alternata and Aureobasidium pullulans were found on the skin of the healthy subjects. When the fungal microbiota of the samples from patients with mild/moderate to severe AD and healthy individuals were clustered together by principal coordinates analysis they were found to be clustered according to health status.     

Immunotherapy in the complex treatment of patients with atopic dermatitis with sensitization to house dust mites

Ninety-nine patients aged 16-52 years with disseminated atopic dermatitis at the remission were examined and treated. The patients were divided into 3 groups according to the method of therapy. Group 1 (28 patients aged 17-52 years, found to be sensitive to Dermatophagoides pteronyssinus and D. farinae) received, in addition to standard treatment, immunotherapy (oral hyposensitization with mite allergens) in combination with ointment containing retinopalmitate and methyluracil. Group 2 (39 patients aged 17-40 years) received standard treatment combined with the administration of placebo. Group 3 (32 patients aged 16-27 years) received only standard therapy. Groups 2 and 3 were used for comparison. The results of treatment were evaluated according to changes in the immune status of the patients and a complex of clinical characteristics. Essential improvement in clinical characteristics and the normalization of immunological parameters were registered in group 1, which proved that immunotherapy was effective and safe.     

Biofeedback treatment of atopic dermatitis

To investigate the feasibility of a behaviorally oriented intervention program with atopic dermatitis, 12 patients were exposed to a fixed sequence of treatment phases including a no-treatment baseline phase, a phase incorporating nonspecific treatment factors, and a phase involving frontal electromyographic (EMG) feedback and relaxation instructions. Photographic analyses of involved skin areas revealed significant remission of dermatological problems across the entire program, although significant changes could not be attributable to any specific phase. Ratings of itching level decreased within but not across treatment sessions, and variable correlations across subjects were found between frontal EMG and itching level. MMPI results from the dermatitis subjects were within normal limits. Overall, the results provided mixed support for the hypothesis that atopic dermatitis may be amenable to intervention through behaviorally oriented treatment procedures.     

Quantitative analysis of cutaneous malassezia in atopic dermatitis patients using real-time PCR

We quantified the cutaneous Malassezia in patients with atopic dermatitis using a real-time PCR assay. Seven to 12 times more Malassezia colonized the head and neck compared to the trunk or limbs, and the species M. globosa and M. restricta accounted for approximately 80% of all Malassezia colonization at any body site.     

Low-energy diet in atopic dermatitis patients: clinical findings and DNA damage

Undernutrition without malnutrition (low-energy diet) increases maximum longevity, reduces the incidence of several cancers and delays their onset, in animal studies. It has also been demonstrated by experimental study that caloric restriction provides a beneficial effect on various inflammatory diseases. In this study, we offered a low-energy diet to patients with atopic dermatitis (AD). Nineteen adult patients (5 males and 14 females aged 15 to 36 years) were enrolled in the study which lasted 8 weeks. The energy intake was 55% of nutritional requirements; protein was 75%, calcium 180%, iron 130%, vitamin A 105%, vitamin C 250% and vitamin E 110% of the daily requirement. No patient experienced adverse reaction, and none dropped out of the trial. Body weight, body mass index (BMI), and systolic blood pressure had decreased significantly by the end of study. The SCORAD (scoring atopic dermatitis) index, which combines objective (extent and intensity of lesions) and subjective (daytime pruritus and sleep loss) criteria, was reduced significantly. In 11 patients with severe AD, there was a significant reduction in oxidative DNA damage. The change in the inflammatory intensity score and the change in BMI caused by energy restriction showed a significant positive correlation. The change in oxidative DNA damage levels and the change in BMI showed a positive correlation. These results clarify the relationship between weight loss and the improvement of AD. It may be hypothesized that this low-energy diet which included several additional nutrients has a possibility to reduce inflammatory symptoms of patients with AD.     

Two-dimensional electrophoretic profiling of atopic dermatitis in primary cultured keratinocytes from patients

Recently, we reported altered protein expression in primary cultured fibroblasts from atopic dermatitis (AD) patients. As a sequential study, we conducted proteomic analysis of primary keratinocytes derived from AD patients to further identify AD-related proteins. Three pH ranges, 4-7, 6-9, and 7-11, were used to profile the altered protein expression in AD. We obtained 46 candidate spots from the 2-D gel image analysis: 18 proteins were up-regulated and 27 proteins were down-regulated. Among the several important candidate proteins, NCC27 showed the same profile of a defect in PTM in both AD-derived keratinocytes and fibroblasts. On the basis of current and previous reports, real-time PCR was performed on select candidate genes to compare RNA and protein expression levels in AD-derived keratinocytes and fibroblasts. Our results provide new clues to aid in understanding the mechanism of atopic alterations in keratinocytes and suggest new AD-associated proteins that are important in AD pathogenesis.     

Nickel allergy and relationship with Staphylococcus aureus in atopic dermatitis

BackgroundThe increase of nickel air pollution is supposed to frequent side effects of nickel action related to virulence potential of Staphylococcus aureus in patients with nickel allergy in atopic dermatitis. The goal was to investigate the relationship between nickel allergy and infection by S. aureus in atopic dermatitis.MethodsNickel allergy was confirmed in atopic patients and excluded in healthy volunteers using patch testing. Infection by S. aureus was tested in atopic patients and healthy volunteers by use of API Staph system. The specific IgE for staphylococcal enterotoxin A and B were measured. Secretion of IFN-g, IL-2, IL-13 by PBMC under nickel sulfate and the enterotoxins A and B stimulations were studied with ELISpot.ResultsWe found the increased number of infections by S. aureus in atopic patients with nickel allergy in comparison to atopic patients and healthy volunteers without nickel allergy. The elevated secretion of IL-2 under nickel sulfate stimulation in vitro was exclusively found in atopic patients with nickel allergy infected by S. aureus.ConclusionsOur data suggest that nickel allergy and infection by S. aureus are linked in atopic dermatitis.     

Recognition of self-heat shock protein 60 by T cells from patients with atopic dermatitis

Heat shock protein 60 (hsp60) is a highly conserved stress protein and target of self-reactive T cells in various inflammatory diseases. Not much is known about a possible role in atopic disease. As atopic diseases are considered to be the result of a disturbance in the balance between T helper cells type 2 and regulatory T cells, it is of interest to know whether hsp60 acts as a bystander antigen in atopic disease. Our aim was to investigate whether hsp60 is involved in the chronicity of inflammation of atopic dermatitis (AD). We studied the expression of hsp60 in skin tissue of adults with AD by immunohistochemistry. Peripheral blood mononuclear cells (PBMC) of children with AD were cultured with hsp60 and proliferative responses, cytokine secretion, surface markers, and functional assays were compared to responses of PBMC of healthy controls (HC). Hsp60 was detected more in lesional skin of AD patients compared to nonlesional skin. Furthermore, PBMC of children with AD proliferated more strongly in response to hsp60 compared to HC. hsp60-reactive T cells of atopic children produced high levels of IFNγ and low levels of IL-10. In vitro activation with hsp60 leads to the induction of CD4⁺CD25^bright T cells expressing FOXP3 in both HC as well as in atopic children. However, despite their regulatory phenotype, hsp60-induced CD4⁺CD25^brightCD127⁻FOXP3⁺ T cells of AD patients were incapable of suppressing effector T cells in vitro. hsp60 is recognized by proinflammatory (IFNγ high, IL-10 low) T cells in atopic patients and is more present in lesional AD skin. This suggests that hsp60-specific T cell responses contribute to local inflammation in AD.

Electronic supplementary material

The online version of this article (doi:10.1007/s12192-012-0361-3) contains supplementary material, which is available to authorized users.     

Role of dysregulated apoptosis in atopic dermatitis

Atopic dermatitis is a chronic inflammatory skin disease with a complex immune dysregulation and interplay of genetic, environmental and psychological factors. Activation and skin-selective homing of peripherial-blood T cells, and effector functions in the skin, represent sequential events in the pathogenesis. Dysregulated apoptosis in skin-homing T cells, eosinophils and keratino-cytes contributes to the elicitation and persistence of atopic derrmatitis.