Isolation of α-toxin-producing Staphylococcus aureus from the skin of highly sensitized adult patients with severe atopic dermatitis

Background  Staphylococcus aureus (S. aureus) is a well-known trigger factor of atopic dermatitis (AD). Besides staphylococcal superantigens, α-toxin may influence cutaneous inflammation via induction of T-cell proliferation and cytokine secretion.
Objectives  To investigate the association between sensitization to inhalant allergens and skin colonization with α-toxin-producing S. aureus in AD.
Patients and methods  We investigated 127 patients with AD, aged 14–65 years, who were on standard anti-inflammatory and antiseptic treatment before investigation. We evaluated skin colonization, medical history, severity of AD and sensitization to inhalant allergens.
Results  Forty-eight of 127 patients were colonized with S. aureus , suffered from more severe AD, had asthma more often and showed higher sensitization levels to inhalant allergens. Thirty of 48 patients with S. aureus skin-colonizing strains produced α-toxin and had higher total IgE and specific IgE to birch pollen and timothy grass pollen.
Conclusions  Under topical treatment with antiseptic and anti-inflammatory agents the colonization of lesional skin with S. aureus was clearly lower than commonly found in untreated patients with AD. Colonization with S. aureus was associated with a higher severity of AD, higher degree of sensitization, and a higher frequency of asthma. The proportion of patients whose skin was colonized with α-toxin-producing S. aureus was higher than expected from a former study. Cutaneous colonization with α-toxin-producing S. aureus was associated with a higher sensitization level to birch pollen allergen in AD. This may point to a higher susceptibility of patients with higher T-helper 2 polarization towards α-toxin-producing S. aureus .     

IgE antibodies to Pityrosporum ovale in atopic dermatitis

An enzyme-linked immunosorbent assay (ELISA) was developed to assess serum IgE antibodies directed against Pityrosporum ovale in patients with atopic dermatitis (AD), atopic patients with allergic respiratory disease (ARD: rhinitis or asthma) but without eczema, and in healthy controls. IgE binding to P. ovale extract was demonstrated in 49% (35/72) of AD patients. In contrast, anti-P. ovale IgE was found in only one of 27 atopic controls without eczema; all healthy control sera (n = 17) were negative. Of 37 AD patients tested intracutaneously with P. ovale, 31 showed immediate-type reactivity, and 20 of these 31 patients had anti-P. ovale IgE detectable by ELISA, while sera from the six non-responders were all negative. Levels of anti-P. ovale IgE were highest in AD patients aged 20-30 years. No correlation was found with the severity of AD, but there was a non-significant tendency (P = 0.06) to higher levels in AD patients with concomittant respiratory allergy. Anti-P. ovale IgE was significantly correlated with total serum IgE, with specific IgE against various aeroallergens as measured by RAST, and with levels of anti-Candida albicans IgE, measured with a similar ELISA. Thus, production of IgE antibodies against P. ovale occurs very frequently in AD, and rarely in patients with atopic disease without skin involvement.     

Serum IgE reactivity to Malassezia furfur extract and recombinant M. furfur allergens in patients with atopic dermatitis

IgE reactivity to the opportunistic yeast Malassezia furfur can be found in patients with atopic dermatitis (AD). We have previously cloned and expressed 6 recombinant allergens (rMal f 1, rMal f 5-9) from M. furfur. In the present study, we used ImmunoCAP to investigate whether these rMal f allergens can be useful in the diagnosis of M. furfur-associated AD compared with the M. furfur extract. A total of 156 adult patients with a clinical diagnosis of AD participated in the study. Sixty-four percent had increased total serum IgE levels, 79% had specific IgE antibodies to common inhalant allergens and 47% had IgE antibodies to M. furfur extract. IgE antibodies to any of the rMal f allergens were detected among 86 (55%) of the patients, 14 (16%) of whom did not react to the M. furfur extract. Any individual rMal f allergen detected between 32% and 89% of the patients ImmunoCAP-positive to the M. furfur extract, with the highest sensitivity for rMal f 9. Therefore, a couple of individual rMal f allergens can improve the diagnosis of M. furfur-associated IgE allergies in patients with AD.     

Polymorphisms within the C3 gene are associated with specific IgE levels to common allergens and super-antigens among atopic dermatitis patients

Abstract:  Atopic dermatitis (AD) is a chronic inflammatory skin disease. Twin and family studies suggest a strong genetic component of the disease. The keratinocytes secrete high amounts of C3 after stimulation with pro-inflammatory cytokines, which may play a functional role in skin inflammation. In this study, we genotyped four different single nucleotide polymorphisms (SNPs) by melting curve analysis using sequence specific hybridization probes in a well-characterized cohort of AD patients. Among four SNPs within C3 gene, higher frequencies of rs10410674 (23.5% vs 12.2%) and rs366510 (13.8% vs 6.5%) were observed in AD patients as compared with control group. None of the tested polymorphisms showed significant association with the risk of the disease phenotype. Analysis of rs10402876 SNP revealed its association with less severe AD disease expression (low SCORAD). Total serum IgE levels were not different among AD patients having any of the four SNPs. However, we observed significantly less serum-specific IgE levels to common allergens ( Dermatophagoides pteronyssinus and birch pollens) and Staphylococcal enterotoxin B in AD patients having rs366510 SNP. Thus, associations of polymorphism within C3 gene with less severe AD disease expression and a weaker sensitization to common allergens suggest the role of these SNPs in the development of AD.     

Ketoconazole in atopic dermatitis: therapeutic response is correlated with decrease in serum IgE

The prevalence of specific IgE antibodies to the yeasts Pityrosporum orbiculare and Candida albicans was investigated in adult patients with atopic dermatitis (AD) or with seborrhoeic dermatitis and in healthy controls by means of the radioallergosorbent test (RAST). Of 63 AD patients, 28 (44%) had IgE antibodies to P. orbiculare and 21 (33%) to C. albicans. This is highly significant, since no antibodies were found in sera from other patients or controls. With the intention to treat, 20 patients with AD and a positive RAST to P. orbiculare were given ketoconazole 200 mg daily for 2 months and 200 mg twice weekly for further 3 months. The clinical scores improved during treatment with a reduction in the levels of specific IgE to P. orbiculare and total serum IgE. However, there were no correlations between clinical score and serum levels of P. orbiculare-specific IgE. C. albicans-specific IgE, on the other hand, correlated both with clinical score and with total serum IgE.     

IgE antibodies to Malassezia furfur, M. sympodialis and Pityrosporum orbiculare in patients with atopic dermatitis, seborrheic eczema or pityriasis versicolor, and identification of respective allergens

Malassezia yeasts may be a trigger factor for atopic dermatitis. Following the recent reclassification of the genus, the presence of specific IgE antibodies was examined in the sera of patients with atopic dermatitis (n = 223), pityriasis versicolor (n = 83), seborrheic eczema (n = 50) and hymenoptera allergy (n = 39) and in controls without skin diseases (n = 50). In addition to using the commercially available radioallergosorbent test (RAST) for Pityrosporum orbiculare couplings were also made against the reference strains for M. furfur and M. sympodialis. To characterize the specificity and molecular weight of corresponding epitopes identical material was used for production of an immunoblot. Despite high total levels of IgE, controls and patients with pityriasis versicolor showed no specific IgE antibodies. Six patients (12%) with seborrheic eczema were positive while 78 patients (35%) with atopic dermatitis had specific IgE antibodies in higher RAST classes that differed between the Malassezia species. The molecular weights of the main antigens of M. sympodialis and M. furfur were determined to be 15, 22, 30, 37, 40, 58, 79, 92, 99 and 124 kDa and 15, 25, 27, 43, 58, 92, 99 and 107 kDa, respectively. Evaluated according to the location of their disease, patients with head and neck lesions most frequently showed Malassezia-specific IgE antibodies. However, there were differences between the Malassezia species tested, the previously used strain P. orbiculare being assignable to the species M. sympodialis.     

Radioimmunologic determination of total IgE and allergen-specific IgE-antibodies in diffuse neurodermitis]

Total IgE levels in sera of 165 patients with atopic dermatitis and 79 patients with dermatoses as well as normal control patients were determined by radioimmunoassay (Phadebas, Pharmacia). Although the mean value for patients with atopic dermatitis was found far above the mean value for normal controls, 38% of patients showed total IgE serum levels within the normal range. Highest IgE serum levels were observed in patients with the generalized form of the disease and in patients with asthma and allergic rhinitis. No direct correlation however, to severity of disease could be found. In a series of 50 patients prick tests were compared to total IgE serum levels and to levels of allergen specific antibodies determined by radioallergosorbent-test (RAST). Extracts of grass pollens and of animal dandruff were used. There was complete agreement between results of skin testing and RAST in at least 80%. While cross reactions were common with grass pollen extracts in RAST, there was no cross-over with animal dandruff. No correlation was found between titer of allergen specific antibody and severity of skin lesions. IgE specific antibody could be detected in 48% of patients with normal total IgE serum levels and in 82% of patients with elevated values.     

Evaluation of RAST in atopic dermatitis

Eighty-two patients with atopic dermatitis were examined for specific IgE antibodies by RAST. A higher number of positive RAST reactions were found in patients with high serum IgE values and with coexisting asthma and/or allergic rhinitis. Positive RAST reactions against mite and house dust were found in many of these patients and even in about one-half of patients with atopic dermatitis only. The comparisons between intradermal tests and RAST were made with various allergens and the mite extract showed the best agreement between these two tests.     

Injection of allergen-antibody complexes is an effective treatment of atopic dermatitis

Atopic dermatitis (AD) can be exacerbated by contact with airborne allergens, amongst which Dermatophagoides pteronyssinus (Dpt) appears to be potentially important. Specific IgE antibodies towards Dpt are often found in AD, and it can therefore be speculated that suppression of the production of anti-Dpt IgE might result in a significant clinical improvement. Complexes of antigen and specific antibodies have been shown to suppress the production of antibody in other systems; we report here the evaluation in an open trial of the capacity of such complexes to improve symptoms of AD. Ten adult patients were enrolled in this study. In addition to satisfying the criteria of AD, they all suffered from a severe disease (more than 20% of the body surface involved) that had been stable for at least the last 2 years. The patients had high titers of total IgE antibodies and specific anti-Dpt antibodies. Allergen-antibody complexes were prepared from Dpt allergens and an excess of autologous specific anti-Dpt antibodies obtained by immunoadsorption. The patients received regular injections of these complexes throughout 1 year, during which clinical parameters of disease intensity, percentage of body surface affected and intensity of pruritus were regularly monitored. A significant clinical improvement was obtained after 3-4 months of therapy and was maintained through the 9th month. After 1 year of treatment, 2 patients were completely free of disease, 4 had residual lesions which continued to improve and 4 patients had a partial recurrence of dermatitis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Staphylococcus aureus-derived enterotoxins enhance house dust mite-induced patch test reactions in atopic dermatitis

BACKGROUND: Up to 65% of Staphylococcus aureus strains isolated from the skin of patients with atopic dermatitis (AD) produce exotoxins with superantigenic properties that may also act as allergens leading to an induction of exotoxin-specific IgE antibodies. Staphylococcal enterotoxin B (SEB) applied epicutaneously in a concentration of 10 micro g/cm(2), i.e. 200 micro g/ml, under occlusion induces cutaneous inflammation in patients with AD and healthy individuals. METHODS: We performed patch tests in 32 adult patients with AD using different concentrations (i.e. 2-200 micro g/ml) of SEA, SEB and house dust mite (HDM) extract (500 micro g/ml). Furthermore, the respective enterotoxins and HDM extract were applied simultaneously to the same patch test site. Specific IgE levels to SEA, SEB and HDM were measured with the CAP FEIA. RESULTS: The rates of patch test reactions to SEA and SEB increased with rising enterotoxin concentrations. There were no differences in the rates of patch test reactions to SEA and SEB between patients sensitized to the corresponding enterotoxin and non-IgE-sensitized patients. The number of patch test reactions to the mixture of enterotoxin and HDM extract was higher than the number of patch test reactions to either the enterotoxins or HDM extract. We identified 11 patients with AD who reacted neither to the enterotoxins nor to HDM extract, but who showed patch test reactions to the mixture. These reactions were not predicted by the presence of enterotoxin- or HDM-specific IgE. CONCLUSIONS: Colonization with exotoxin-producing S. aureus may influence the outcome of patch tests in patients with AD.     

Pityrosporum ovale and atopic dermatitis in children and young adults.

Children aged 0-21 years, 60 children with atopic dermatitis (AD), 40 children with rhinoconjunctivitis and or asthma (RA) and 40 children with no atopic history (HC) were studied to evaluate the relationship between skin colonisation with Pityrosporum ovale and the occurrence of specific IgE antibodies to P. ovale. The following studies were done: culture for P. ovale, measurement of IgE antibodies to P. ovale (skin prick test, RAST), Candida albicans, and Cladosporium herbarum (RAST) and IgG antibodies to P. ovale. P. ovale could be cultured with about the same frequency in children and young adults with AD and age-matched children with or without other atopic manifestations. In spite of similar colonisation, IgE antibodies against P. ovale occur only in atopy and more frequently in children with AD than in those with other types of atopic disease.     

Atopic disease and serum immunoglobulin-E

Fifty-seven subjects, forty-three with various combinations of atopic disease and fourteen non-atopic controls, were studied using a battery of immediate skin test allergens and a radio-immunoassay for serum Immunoglobulin-E (IgE). The geometric mean serum IgE level (units/ml) was 50 in strictly nonatopic controls, 170 in atopic respiratory disease (ARD) patients, 320 in atopic dermatitis (AD) patients and 772 in those patients with both AD-ARD. The marked elevation of the serum IgE in the latter group was not associated with respiratory disease activity and not closely correlated with extent of the dermatitis, but may relate to patients with both AD-ARD being 'highly atopic'. Overall, there was no correlation between the serum IgE level and the frequency of positive skin tests. Compared to ARD patients, the serum of AD patients contained more IgE, yet they reacted significantly less frequently to common extrinsic allergens.     

Prevalence of atopic dermatitis and serum IgE values in nursery school children in Ishigaki Island, Okinawa, Japan

There have been many studies of the prevalence of atopic dermatitis (AD), but few population-based epidemiologic studies measure the prevalence in Japan among children aged 5 years and younger. We examined the prevalence of AD, serum total IgE levels and specific IgE antibodies to 10 common allergens among children in Ishigaki Island, Okinawa, Japan in 2001. We also obtained information on the predictability of the U.K. Working Party diagnostic questionnaire criteria for AD in this population. Five hundred and sixty five children aged 5 years and younger were enrolled in this study with informed consent from their parents. The questionnaire of the U.K. Working Party diagnostic criteria for AD was translated into Japanese, and the parents completed the questionnaire sheet. Physical examination and blood sampling were done for all children. Thirty-nine out of the 565 (6.9%) children were diagnosed with AD by physical examination. The total and specific IgE levels were significantly higher in the children with AD than in those without AD. High levels of total IgE were found in 33.3% of the children with AD. A specific IgE to one or more allergens was detected in 64.1% of children with AD. However, a substantial population of children without AD also had high levels of total IgE (12.7%) and a specific IgE to one or more allergens (30.2%), and the increment of total and specific IgE levels was significantly associated with age. The percentage of positive answers to the questionnaire of the U.K. Working Party diagnostic criteria for AD was significantly higher in children with AD (59.0%) than in children without AD (5.3%) (P<0.0001). Its specificity was 94.7%. The false negative rate was 41%. In conclusion, the prevalence of AD was relatively low in children in Ishigaki Island. High levels of total IgE were found in only one third of children with AD under 5 years of age. The Japanese translated form of the questionnaire of the U.K. Working Party diagnostic criteria for AD should be refined to improve its sensitivity.     

Levels of immunoglobulin E specific to the major food allergen and chemokine (C‐C motif) ligand (CCL)17/thymus and activation regulated chemokine and CCL22/macrophage‐derived chemokine in infantile atopic dermatitis on Ishigaki Island

Atopic dermatitis (AD) is a multifactorial T‐helper (Th)2‐mediated skin disease frequently associated with elevated serum immunoglobulin (Ig)E and food allergy is also a Th2‐ and IgE‐mediated adverse immunological reaction. Our previous study indicated the relation of egg allergy history and disease severity of AD. Thus, the purpose of the study was to investigate the levels of IgE specific to major food allergens (egg, milk, wheat) and Th2 chemokines (chemokine [C‐C motif] ligand [CCL]17/thymus and activation regulated chemokine [TARC] and CCL22/macrophage‐derived chemokine [MDC]) and the relationship between them. A total of 743 nursery school children were enrolled. Dermatologist‐based physical examination and a questionnaire survey were also conducted. Significantly increased levels of disease severity markers (CCL17/TARC and CCL22/MDC) were confirmed in children with AD. The levels of CCL22/MDC in all of the children were markedly high compared with those reported in adults. IgE specific to egg white, ovomucoid, wheat and mite antigen were significantly higher in the AD group than in the non‐AD group. Among them, IgE specific to egg allergens were well associated with disease severity markers, and IgE specific to ovomucoid seemed particularly well correlated with the presence of egg allergy history. In conclusion, the markedly high level of CCL22/MDC in children as compared with those reported in adults may partly explain the AD‐prone nature of children and their spontaneous remission afterwards. Mild but significant correlation of IgE specific to egg allergens and Th2 chemokines may explain correlation of disease severity and comorbidity of egg allergy in our previous study.     

The prevalence of antibody responses against Staphylococcus aureus antigens in patients with atopic dermatitis: a systematic review and meta‐analysis

Atopic dermatitis (AD) is a common type of eczema. The bacterium Staphylococcus (S.) aureus plays a role in AD, possibly via various substances that are part of the bacterium. In people with AD, how the body's immune system reacts to these substances might lead to the inflammation seen in AD. In this study, we provide an overview of what is known from current literature (e.g. studies published in medical journals) on the immune response (antibodies) against S. aureus substances (antigens) in AD patients compared to healthy controls (people without AD). Scientific literature was systematically obtained from different databases. We extracted data on how often (so‐called ‘prevalence’) antibodies of, among others, the IgE type directed against S. aureus can be found in AD patients. A weighted prevalence was calculated, as studies with more patients were regarded more relevant, and prevalence was compared between AD patients and healthy controls. Tests for differences between studies and overall study quality were done to interpret the robustness of the results. We included 26 publications with a total of 2369 patients. In 10 studies a control group (of people without AD) was also studied. Study quality was fair to poor. The weighted prevalence of IgE against S. aureus antigens was 33% for staphylococcal enterotoxin (SE) A, 35% for SEB and 16% for toxic shock syndrome toxin (TSST)‐1. Compared to healthy controls, AD patients had 8.37 times more IgE against SEA and 9.37 times more IgE against SEB. No significant difference was seen for IgE against TSST‐1. Interpretation of the results should take into account that there were some important differences in study design and the fact that the studies were generally small. In conclusion, AD patients more often show an IgE antibody response against the S. aureus antigens SEA and SEB compared to healthy controls. These findings show that S. aureus may have a role in AD.     

Immune response to Staphylococcus aureus in atopic dermatitis

The skin of patients with atopic dermatitis (AD) is severely colonized with Staphylococcus aureus. Therefore, a study was conducted to assess some basic features of the S. aureus-specific immune response in patients with AD and healthy nonatopic individuals. Some particular features were found: a selective hyporesponsiveness to purified S. aureus cell walls (PCW) in delayed skin reactivity; half of our AD patients showed serum IgE to PCW and soluble S. aureus protoplast antigens; elevated PCW-IgE did not correlate with positive immediate skin reactions to whole S. aureus and their cell walls; regional lymphadenopathy but not impetiginization was associated with increased PCW-IgE and high total IgE. It is suggested that these changes in the immune response to S. aureus are related to the chronic S. aureus colonization of the skin.     

Phenotyping of epidermal dendritic cells allows the differentiation between extrinsic and intrinsic forms of atopic dermatitis

Atopic dermatitis (AD) is a clinically characteristic, chronic inflammatory skin disease of unknown origin. IgE-mediated uptake and antigen focusing of environmental allergens by dendritic cells (DCs) is assumed to be a central immunopathogenetic event. A so-called intrinsic type of AD (IAD) has been delineated from the more common extrinsic AD (EAD) by normal serum IgE levels, negative RAST tests and negative immediate-type skin reactions towards environmental allergens. The recently characterized human autoantigen Hom S 1 has been proposed to play a part in the pathogenesis of IAD. OBJECTIVES: To compare clinical and laboratory data between patients with IAD and EAD, and to investigate potential differences in the inflammatory micromilieu of the epidermal compartment in IAD and EAD lesions. METHODS: Epidermal DC phenotyping, a recently validated technique based on the three-colour flow cytometric analysis of Langerhans cells and the so-called inflammatory dendritic epidermal cells from epidermal single-cell suspensions, was performed on samples from 69 patients with AD (seven with IAD and 62 with EAD) and 94 controls. RESULTS: Patients with EAD tended to have an earlier onset of disease but similar disease duration and family history of atopic diseases. Quantitative analysis of CD36 expression on DCs as a marker of inflammation, as well as the percentage of inflammatory dendritic epidermal cells in the CD1a+ epidermal DC pool, indicated a comparable disease activity in IAD and EAD. EAD was characterized by a significantly higher FcepsilonRI expression on the CD1a+ epidermal DCs than IAD. Using the FcepsilonRI/FcgammaRII expression ratio as a disease marker for AD, values for IAD fell below the diagnostic cut-off level of 1.5 for this ratio. CONCLUSIONS: While IAD is clinically similar to EAD, the inflammatory microenvironment in this condition seems different from classical EAD and can be distinguished by phenotyping of epidermal DCs.     

Most common inhalant allergens in atopic dermatitis, atopic dermatitis/allergic rhinitis, and atopic dermatitis/bronchial asthma patients: a five-year retrospective study

Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disease with a distinctive clinical appearance and distribution. Around 85% of patients have positive immediate skin reaction or specific IgE to different airborne allergens that are in association with respiratory allergy. The aim of this retrospective, open and uncontrolled study was to identify the most common inhalant allergens in AD patients, AD/allergic rhinitis patients, and AD/bronchial asthma patients by skin prick test per year during the 2001-2005 period.     

Serum IgE Antibodies to the Scabies Mite

Abstract: Porcine scabies mites, Sarcoptes scabiei var. suis are more readily available in sufficient quantities than are human scabies mites. Circulating IgE antibodies specific to porcine scabies mites were found in 6 (30%) of 20 scabies patients with an RAST score of at least 2. Seven patients had elevated serum total IgE levels. Correlation between the RAST values, the duration of pruritus, and the IgE levels was not found. The results do not prove the existence of antibodies specific to porcine scabies mites but strongly support this assumption.     

Correlation of serum total IgE, eosinophil granule cationic proteins, sensitized allergens and family aggregation in atopic dermatitis patients with or without rhinitis

Atopic dermatitis (AD) is a complex disease with both a genetic background and environmental interactions. Although multiple linkage-analyses about AD have been studied, there have been only a few family aggregation tests of AD or perennial allergic rhinitis (AR) to date. The association of allergen-specific IgE in AD and atopic dermatitis with allergic perennial rhinitis (ADR) have also been seldom discussed. The purpose of this study was to evaluate family aggregation and assess allergen-specific IgE in patients with AD and ADR. We also planned to investigate the effect of family history of AD on the prevalence of allergen-specific antibodies. The serum levels of IgE, eosinophil cationic protein (ECP) and major basic protein (MBP) were measured and compared in patients with AD and those with ADR. Proportional analysis compared allergen-specific IgE between AD and ADR. The family aggregation was conducted to estimate the odds ratio for various atopic diseases in different family members. Total IgE and allergen-specific antibodies in serum were compared between those patients who had AD with AR and those without. The result revealed that allergic rhinitis is the most common concomitant atopic disease associated with AD. The ADR group was more likely to have serum mite-, cockroach-, and feather-specific IgE. The positive rates for wheat, peanut and soybean were higher in those AD without rhinitis. In the family aggregation of AD, the odds ratio for siblings was higher than for parents, the ratios for brother and sister were 9.91 and 8.75, respectively. However, the odds ratio for parents of ADR was higher than siblings; the ratios for father and mother of ADR were 8.22 and 2.94, respectively. AD patients with family histories of AD were more likely to have mite-, soybean-, and peanut-specific antibodies in their serum. We concluded that aeroallergens are the most important allergens aggravating atopic diseases in Taiwan. Food plays an important role in the pathogenesis of AD. Measurement of serum total IgE combined with the MAST-CLA test could be helpful in the diagnosis of atopic diseases. The differential aggregation tendency for AD and ADR implicated the complexity of the gene-environment interaction in these atopic diseases.