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Abstract: Background: Allergen‐specific immunotherapy (SIT) effectively alleviates type I allergic diseases characterized by T helper (Th)2‐type immunity. Our recent studies have shown that a synthetic trivalent glycocluster, triacedimannose (TADM), suppresses the Th2‐type allergic inflammation. The aim of this study was to compare TADM with two well‐known adjuvants, unmethylated cytosine‐phosphate‐guanine oligodeoxynucleotide (CpG) and monophosphoryl lipid A (MPLA) in a grass allergen‐induced chronic allergic inflammation model in mice. Methods: Female BALB/c mice were intranasally sensitized with 50 μL of timothy grass pollen extract (TE) twice a week for a period of 15 weeks. Therapeutic intranasal treatments were then performed once a week after the tenth intranasal TE instillation using TADM (10 or 25 μg/50 μL), CpG‐ODN (20 μg/50 μL) or MPLA (2 μg/50 μL). Groups of 9‐10 animals per treatment were killed 24 hours after the last timothy dosage. Blood, bronchoalveolar lavage (BAL) fluids and lung biopsies were taken for subsequent analysis. Results: When mice were repeatedly exposed to TE for 15 weeks, the number of eosinophils and lymphocytes increased in the BAL fluids. The eosinophil and lymphocyte counts decreased dose‐dependently and were practically abolished in the mice treated with TADM. Treatments with MPLA or CpG significantly increased the number s of neutrophils, while CpG nonsignificantly decreased eosinophilia compared to timothy exposure. Conclusions: A novel synthetic glycocluster molecule inhibited the development of grass‐induced eosinophilic pulmonary inflammation in mice when administrated in the airways. This compound could be a candidate to be used either as an adjuvant in SIT or as a topical anti‐inflammatory treatment. [ABSTRACT FROM AUTHOR]

Background During infancy, a disturbed cytokine balance leads to an atopic immune response. Many risk factors have been associated with the development of atopy. These include parental smoking, elevated cord blood IgE, early exposure to pets and family history of atopy, but the knowledge of their impact on cytokine balance is limited. Objective To assess the cytokines induced by mitogen in peripheral blood mononuclear cells ( PBMC) of infants at 3 months and 12 months of age and their potential association with fatty acid ( FA) intervention, parental atopy, atopic dermatitis and parental smoking. Methods Infants from an intervention study using black currant seed oil ( BCSO, n = 34) or placebo ( n = 34) were included. PBMC samples were taken at the age of 3 and 12 months. Signs of atopic dermatitis and parental smoking were registered. PBMC were isolated from heparinized blood samples, stimulated with ConcanavalinA mitogen and the cytokine responses were detected at 72 h of stimulation by Luminex technology. Results Children of smoking parents had elevated levels of IL-4 ( P = 0.0004), IL-5 ( P = 0.0002), IFN-γ ( P = 0.039) and TNF ( P = 0.0003) at 12 months of age. Children who had atopic dermatitis by the age of 3 months showed elevated levels of IL-5 at 3 months ( P = 0.0027) and 12 months of age ( P = 0.022). The production of TNF at the age of 3 months was higher ( P = 0.010) and the production of IL-12 at the age of 12 months was lower ( P = 0.025) in infants whose parents were atopic. BCSO intervention did not have any effect on any cytokine production or mRNA expression. Conclusion Children of smoking parents had highly significantly elevated levels of Th2-type cytokines IL-4, IL-5 and pro-inflammatory cytokine TNF. The detrimental effects of parental smoking on the child's immune function should lead us to pay more attention to supporting parents to stop smoking. [ABSTRACT FROM AUTHOR]

Background The present increased incidence of atopic diseases has been associated with an altered intake of essential fatty acids (EFAs). The composition of blackcurrant seed oil (BCSO) corresponds to the recommended dietary intake of EFAs, and as a dietary supplement could, in small doses, modify the imbalance of EFAs in an efficient way. Objective To assess the effect of dietary supplementation with BCSO on the prevalence of atopy at 12 months of age. Methods Three hundred and thirteen pregnant mothers were randomly assigned to receive BCSO (151) or olive oil as placebo (162). The first doses were administered at 8th–16th weeks of pregnancy and were continued until the cessation of breastfeeding, followed by supplementation to the infants until the age of 2 years. Atopic dermatitis and its severity (SCORAD index) were evaluated, serum total IgE was measured and skin tests were performed at the age of 3, 12 and 24 months. Results Parental atopy was common (81.7%) among study subjects, making them infants with increased atopy risk. There was a significantly lower prevalence of atopic dermatitis in the BCSO group than in the olive oil group at the age of 12 months (33.0% vs. 47.3%, P=0.035). SCORAD was also lower in the BCSO group than in the olive oil group at 12 months of age ( P=0.035). No significant differences in the prevalence of atopic dermatitis were observed between the groups at the age of 24 months ( P=0.18). Conclusion Dietary supplementation with BCSO was well tolerated and it transiently reduced the prevalence of atopic dermatitis. It could therefore be one potential tool in the prevention of atopic symptoms when used at an early stage of life. (Registration number SRCTN14869647, ) Cite this as: P. Linnamaa, J. Savolainen, L. Koulu, S. Tuomasjukka, H. Kallio, B. Yang, T. Vahlberg and R. Tahvonen, Clinical & Experimental Allergy, 2010 (40) 1247–1255. [ABSTRACT FROM AUTHOR]

In similarity to many other western countries, the burden of allergic diseases in Finland is high. Studies worldwide have shown that an environment rich in microbes in early life reduces the subsequent risk of developing allergic diseases. Along with urbanization, such exposure has dramatically reduced, both in terms of diversity and quantity. Continuous stimulation of the immune system by environmental saprophytes via the skin, respiratory tract and gut appears to be necessary for activation of the regulatory network including regulatory T-cells and dendritic cells. Substantial evidence now shows that the balance between allergy and tolerance is dependent on regulatory T-cells. Tolerance induced by allergen-specific regulatory T-cells appears to be the normal immunological response to allergens in non atopic healthy individuals. Healthy subjects have an intact functional allergen-specific regulatory T-cell response, which in allergic subjects is impaired. Evidence on this exists with respect to atopic dermatitis, contact dermatitis, allergic rhinitis and asthma. Restoration of impaired allergen-specific regulatory T-cell response and tolerance induction has furthermore been demonstrated during allergen-specific subcutaneous and sublingual immunotherapy and is crucial for good therapeutic outcome. However, tolerance can also be strengthened unspecifically by simple means, e.g. by consuming farm milk and spending time in nature. Results so far obtained from animal models indicate that it is possible to restore tolerance by administering the allergen in certain circumstances both locally and systemically. It has become increasingly clear that continuous exposure to microbial antigens as well as allergens in foodstuffs and the environment is decisive, and excessive antigen avoidance can be harmful and weaken or even prevent the development of regulatory mechanisms. Success in the Finnish Asthma Programme was an encouraging example of how it is possible to reduce both the costs and morbidity of asthma. The time, in the wake of the Asthma Programme, is now opportune for a national allergy programme, particularly as in the past few years, fundamentally more essential data on tolerance and its mechanisms have been published. In this review, the scientific rationale for the Finnish Allergy Programme 2008–2018 is outlined. The focus is on tolerance and how to endorse tolerance at the population level. [ABSTRACT FROM AUTHOR]

Background: Signalling lymphocytic activation molecule (SLAM) and interleukin (IL)-18 induce interferon (IFN)-γ production from Th1 cells. The allergen-induced SLAM and IL-18 mRNA expressions are increased during subcutaneous immunotherapy (SCIT), but nothing is known about their role during sublingual immunotherapy (SLIT). Transcription factor GATA-3 is associated with Th2 cells but its role in SCIT and SLIT is yet unexplored. This study was undertaken to analyse the allergen induced in vitro mRNA expression of IL-18, SLAM and GATA-3 in peripheral blood mononuclear cells (PBMC) of children with allergic rhinitis (AR) during SLIT. Methods: Ten patients with AR undergoing pollen SLIT with a weekly dose of 200 000 SQ-U, 10 with 24 000 SQ-U of mixture of Betula verrucosa, Corylus avellana and Alnus glutinosa and 10 with placebo were included. Peripheral blood mononuclear cell were stimulated with birch extract prior to, after 1 and 2 years of the treatment. The mRNA expression was assessed using kinetic real-time RT-PCR (TaqMan®; Applied Biosystems, Foster City, CA, USA). Results: The expression of IL-18 mRNA was increased in the high-dose group in comparison to the placebo group after 1 year of therapy ( P = 0.028) and had an inverse correlation with the late phase skin reaction after the second study year ( r = −0.41, P = 0.041). SLAM mRNA expression increased in the high-dose group from baseline to 1 year ( P = 0.028) and correlated with IL-10 ( r = 0.96, P < 0.0001) and transforming growth factor-β ( r = 0.80, P = 0.0037) mRNA expression. No significant changes were seen in GATA-3 mRNA expression. Conclusions: During SLIT, IL-18 and SLAM are upregulated, suggesting that the Th2 type inflammatory response is downregulated during SLIT by increased Th1 type response. [ABSTRACT FROM AUTHOR]

Background: During subcutaneous immunotherapy (SCIT), there is a local mucosal shift from Th2 to Th1 type cytokine predominance and downregulation of interleukin (IL)-5 and eosinophilia. According to recent studies IL-10- and transforming growth factor (TGF)- β-induced tolerance is another key phenomenon in SCIT. Few data to date is available on mechanisms and roles of these cytokines in sublingual immunotherapy (SLIT). Scope: This study was undertaken to analyse the allergen-induced in vitro mRNA expression of IL-4, IL-5, IL-10, TGF- β and interferon (IFN)- γ during SLIT in peripheral blood mononuclear cells (PBMC) of children with allergic rhinitis (AR). Methods: Ten patients with AR undergoing pollen SLIT with a weekly dose of 200 000 SQ-U, 10 with a weekly dose of 24 000 SQ-U of glycerinated mixture of Betula verrucosa, Corylus avellana and Alnus glutinosa and 10 with placebo were included in the study. Peripheral blood mononuclear cell samples were collected and stimulated with pollen allergen extract prior to the treatment, after 1 and 2 years of the treatment. The cytokine mRNA expression was assessed using kinetic real time reverse transcription polymerase chain reaction (RT-PCR; TaqMan®). Results: The in vitro allergen-induced mRNA expression of IL-5 by PBMC in the placebo group at 1 ( P = 0.0065) and 2 ( P = 0.013) years of therapy were increased in comparison with the highest dose. The expression of IL-10 mRNA was increased in the highest dose group ( P = 0.0016) and the lower dose group ( P = 0.034) at 2 years of therapy when compared with placebo. The change in the expression of allergen-induced TGF- β had an inversed correlation with the change of IL-5 ( r = −0.38, P = 0.036) and positive correlation with the change of IL-10 ( r = 0.58, P = 0.0019). Conclusions: Sublingual immunotherapy induced a dose-dependent systemic allergen-specific immunological response in children with AR. During high-dose SLIT, there was activation of regulatory cytokine IL-10 and an inhibitory effect on IL-5 expression increase that was associated with TGF- β. [ABSTRACT FROM AUTHOR]

Background: Subcutaneous immunotherapy has been the principal approach of immunotherapy in the treatment of allergic diseases. Several clinical studies with birch, alder or hazel pollen extract conducted as subcutaneous immunotherapy have been published suggesting a well-tolerated and clinically effective treatment. Only a few clinical studies of sublingual immunotherapy (SLIT) with these allergens have been published. This study investigated the clinical efficacy, safety and dose–response relationship of SLIT in children suffering from rhinoconjunctivitis with/without asthma. Methods: Eighty-eight children (5–15 years) with a history of tree pollen-induced allergic rhinoconjunctivitis with/without seasonal asthma for ≥2 years were included. Allergy to tree pollen was confirmed by positive skin-prick test, positive specific IgE and positive conjunctival provocation test. The extract used was a glycerinated mixture of Betula verrucosa, Corylus avellana and Alnus glutinosa 100 000 SQ-U/ml. Children were randomized into three groups receiving SLIT 5 days a week for up to 18 months; dose group 1: accumulated weekly dose of 24 000 SQ-U; dose group 2: accumulated weekly dose of 200 000 SQ-U; and placebo. Results: In the birch pollen season, dose group 2 showed a significant reduction of symptom ( P = 0.01) and medication scores ( P = 0.04) compared with placebo. Dose group 1 showed a significant reduction of symptom scores ( P = 0.03). There were no statistical differences between dose groups 1 and 2. All children tolerated the treatment well. Conclusion: SLIT with tree pollen extract provided dose-dependent benefits in tree pollen-allergic children in terms of significantly reduced symptoms and medication use. The treatment was well tolerated. [ABSTRACT FROM AUTHOR]

Elevated and correlativeMalassezia furfur(M. furfur) andCandida albicans(C. albicans) mannan-specific IgE have been demonstrated in atopic eczema dermatitis syndrome (AEDS) of the head, neck and shoulder (HNS) region of the skin. The significance of these antibodiesin vivohas not been demonstrated.Sixty-five AEDS patients with HNS distribution were included. Serum total IgE (S-IgE) and yeast antigen-specific (Cetavlon-purified mannan and whole extract antigens ofM. furfurandC. albicans) IgE were measured and skin prick tests (SPT) were performed with the yeast antigens.Mannan-specific IgE and SPT were positive in 51 and 48% of patients withM. furfurand in 42 and 22% withC. albicans, respectively. Whole extract-specific IgE and SPT were positive in 85 and 95% of patients withM. furfurand in 91 and 57% withC. albicans, respectively. The highest correlation between specific IgE and SPT was seen withM. furfurmannan (r = 0.60;P < 0.0001). BothM. furfurmannan-specific IgE (r = 0.76;P < 0.0001) and SPT (r = 0.44;P = 0.0005) correlated with S-IgE.Mannan-induced immediate hypersensitivityin vivowas demonstrated in SPT. The significant correlation betweenM. furfurmannan-specific IgE and SPT suggests that mannan is an important allergen in yeast hypersensitive AEDSin vivo. [ABSTRACT FROM AUTHOR]

T-box expressed in T cells (T-bet) is a transcription factor regulating the commitment of T helper (Th) cells by driving the cells into the Th1 direction. Abnormal Th1/Th2 balance may lead to complex disorders like asthma or autoimmune diseases. Recent studies have suggested that T-bet might be a candidate gene for asthma. This led us to screen 23 Finnish individuals for single-nucleotide polymorphisms (SNPs) in the T-bet locus and study the association between the SNPs and high serum IgE level and asthma. We screened all six exons, adjacent intronic areas and 2 kb of the 5′-flanking region from 23 individuals utilizing WAVE™ technology. To explore whether T-bet is associated in serum IgE regulation or asthma we genotyped the SNPs in a Finnish asthmatic founder population. The association analyses were made using haplotype pattern mining. Fifteen novel SNPs were found in the T-bet gene. Within the Finnish asthmatic founder population, there was no association between T-bet SNPs and high serum IgE level or asthma. The genetic variability in the T-bet gene does not play a role in the pathogenesis of human asthma. Our results provide a novel panel of SNPs in T-bet and will help determine whether the SNPs have a functional role in other T cell-mediated diseases. [ABSTRACT FROM AUTHOR]

During specific pollen immunotherapy (SIT) there is a local mucosal shift from Th2- to Th1- type cytokine predominance, with IL-12 having a major role in this shift. IL-10-induced tolerance is supposed to be a key phenomenon in venom immunotherapy (VIT). However, the role of Th1-promoting cytokines, on the one hand, and the role of regulatory cytokines, on the other hand, have not been studied in parallel during SIT. This study was undertaken to analyse the allergen-induced in vitro mRNA expression of Th1-type effector cytokine IL-18 and regulatory cytokines IL-10 and TGF-β during SIT in peripheral blood mononuclear cells (PBMC) of allergic rhinitis (AR) patients. Thirty patients with AR undergoing pollen SIT and 10 patients with AR who were not treated with SIT were included in the study. The symptoms and medications were registered post-seasonally before the beginning of SIT and after 1 year of therapy. PBMC samples were collected and stimulated with pollen allergen extract prior to the treatment, at the maintenance phase in 12 patients and after 1 year of the treatment. The cytokine mRNA expression was assessed using kinetic real-time RT-PCR (TaqMan®). There was a clear increase in the treated AR patients, in comparison with untreated AR patients, in the expression of both IL-10 (mean change from baseline (SEM): 3.1 (0.8) vs. −0.3 (0.3), P<0.002, Mann–Whitney U-test) and IL-18 (2.7 (0.9) vs. −0.2 (0.6), P<0.03) mRNA after 1 year. The clearest increase in IL-10 mRNA expression was seen in patients who did not benefit at all (6.0 (2.3), P<0.001 vs. untreated) and the least increase in patients that had the greatest reduction of symptoms (0.8 (0.6), n.s. vs. untreated) at 1 year. The clearest increase in IL-18 mRNA expression was seen in patients with moderate outcome (3.4 (1.6), P<0.04 vs. untreated). In intermediate samples, taken when the maintenance dose was reached, the peak expression of allergen-induced IL-10 mRNA was associated with the most favourable outcome of SIT ( P=0.01, Fisher exact test). A similar trend was seen in IL-18 mRNA expression. The results suggest that an early and transient increase in allergen-specific IL-10 and IL-18 mRNA expression in PBMC is essential for the therapeutic outcome after 1 year of SIT. [ABSTRACT FROM AUTHOR]

Atopic eczema/dermatitis syndrome (AEDS) patients display immunoglobulin E (IgE) reactivity to several antigens, e.g. saprophytic yeasts as Malassezia furfur. AEDS patients also show IgE autoreactivity towards cells of their own tissue including epidermis. The aim of this study was to investigate the IgE autoreactivity of AEDS patients to cultured keratinocytes and to reveal potential crossreacting epitopes in cultured keratinocytes and M. furfur. Serum samples of 27 AEDS patients were analyzed, of these 13 were M. furfur radioallergosorbent test (RAST) positive and 14 negative. Four urticaria, three psoriasis, and seven nonatopic patients were included as controls. The studies were performed by using IgE immunoblotting and immunoblotting inhibition methods. Ten IgE-binding protein bands were detected in cultured human keratinocytes by IgE immunoblotting using sera from adult AEDS patients. Anti-keratinocyte IgE antibodies were more associated with elevated S-IgE level than M. furfur RAST. Clear crossreactivity with M. furfur could not be shown. The possible pathomechanism of anti-keratinocyte IgE antibodies is not due to IgE epitope mimicry of saprophytic yeast and local tissue in AEDS skin. [ABSTRACT FROM AUTHOR]

Background: In yeast-sensitive atopic eczema dermatitis syndrome (AEDS), yeast mannan induces highly elevated specific IgE levels and lymphoproliferative responses. In healthy individuals the involvement of both human leukocyte antigen (HLA)-dependent T-cell activation and non-HLA-dependent activation, e.g. by crosslinking of the cell surface mannose receptors, has been suggested. In the present study the HLA dependence and the role of crosslinking in the lymphoproliferative response to mannan in AEDS has been analyzed. Methods: Twenty patients with AEDS and 12 controls with no history of allergic diseases were included in the study. Mannan from Candida albicans was prepared according to the Cetavlon method. Following isolation using Ficoll-Hypaque, peripheral blood mononuclear cells (PBMC) were incubated with the mannan preparation in the absence and presence of different concentrations of neutralizing anti-HLA antibodies and α-methylmannoside for 6 days and proliferative responses were measured by 3 H-thymidine incorporation and scintilloradiography. Results: In AEDS patients with elevated mannan-specific serum IgE, the C. albicans mannan induced lymphoproliferation. Mannan-induced lymphoproliferative responses could be inhibited, dose-dependently, by neutralizing anti-HLA-DR, but not anti-HLA-DQ antibodies in AEDS patients and healthy controls. The addition of α-methylmannoside, that blocks binding to mannose receptors, inhibited lymphoproliferative responses in a dose-dependent way by 50% only in healthy controls, but not in AEDS patients. Levels of inhibition of the proliferation by α-methylmannoside correlated inversely with the yeast- and mannan-specific IgE levels. Conclusions: These results show that in healthy subjects yeast mannan activates lymphocytes both in an HLA-DR-dependent manner and as a result of direct crosslinking of the cell surface. However, in AEDS the elevated lymphoproliferative response is HLA-DR-dependent,... [ABSTRACT FROM AUTHOR]

Background: According to a hypothesis allergens induce Th2 responses in allergic patients, and microbes induce Th1 responses. We studied the kinetics of in vitro allergen-, tuberculin (PPD)- and tetanus toxin (TT)-induced IFN-γ and IL-4 mRNA expression in peripheral blood mononuclear cell (PBMC) cultures of pollen-allergic patients and healthy controls. Methods: PBMC of 10 birch or timothy pollen-allergic patients and of 13 healthy controls were stimulated in vitro with allergen (birch or timothy), PPD or TT. Pellets and supernatants were collected at 24, 48, 72 and 96 h after stimulation. IFN-γ and IL-4 production was measured by enzyme linked immunosorbent assay and mRNA expression using RT-PCR and time-resolved fluorometry. Results: Allergen induced IFN-γ production and mRNA expression in PBMC more in allergic patients than in healthy controls. Also allergen induced IL-4 mRNA expression more in allergic patients than in healthy controls. PPD induced IFN-γ mRNA expression both in allergic patients and healthy controls, whereas IFN-γ production was induced only in healthy controls and IL-4 was not induced at all. TT induced IFN-γ mRNA expression in both groups, IFN-γ production in allergic patients, and IL-4 mRNA expression in both allergic patients and healthy controls. Conclusions: In vitro stimulation with allergen induced both IFN-γ and IL-4 mRNA expression of PBMC in allergic patients. These observations challenge the clearcut division of microbe-specific Th1 and allergen-specific Th2 responses in peripheral blood. [ABSTRACT FROM AUTHOR]

Background: IgE-mediated hypersensitivity to yeasts is often seen in atopic dermatitis (AD) patients, especially when dermatitis is located in the head, neck, and shoulder regions. Two studies have shown the efficacy of ketoconazole in the treatment of this type of AD, in contrast to results of topical treatment. The objective was to assess the clinical efficacy of antifungal treatment in AD in a randomized, double-blind, placebo-controlled study with oral ketoconazole and yeast-specific IgE levels and saprophytic yeast growth monitored simultaneously. Methods: Eighty patients with AD and positive P. ovale and/or C. albicans RAST/skin prick test results were randomized to receive ketoconazole or placebo for 30 days. The yeast growth of skin and pharynx; P. ovale, C. albicans, andS. cerevisiae RAST; serum total IgE; and the severity of the eczema (SCORAD) were assessed at day 0 and thereafter at 1 and 3 months. Results: A significant improvement was seen in the SCORAD scale in the ketoconazole group at the second visit in comparison to the first visit (P<0.0005; n=36), but not in the placebo group (n=39). Of the individual determinants of the SCORAD, itching (P<0.005), the extent of dermatitis (area percentage), excoriation, lichenification (P<0.01), erythema, papulation, and dryness (P<0.05) improved significantly in the ketoconazole group. In the placebo group, only the extent of dermatitis (area percentage) decreased significantly (P<0.05). In the ketoconazole group, the number of positive P. ovale cultures decreased from 60% to 31% (n=35) compared to the placebo group (64% to 56%; n=39). The clinical response was most significant in female patients with positive yeast cultures. Conclusions: Saprophytic yeasts may be a source of allergens in AD. Thus, patients with AD, yeast growth, and elevated IgE levels to yeasts should be offered antifungal treatment. [ABSTRACT FROM AUTHOR]

Recent cytokine (RT-PCR, ELISA) analyses of inflammation in atopic dermatitis (AD) have suggested a role for IL-4, IL-5 and IFNγ. Pityrosporum ovale and Candida albicans are important allergens in some patients with AD of the seborrhoic head, neck and shoulder region. In AD patients, the saprophytic yeasts induce IgE responses while they usually induce TH[sub 1] type responses. The cytokine responses induced by yeasts in AD are sparsely investigated. To characterize the P. ovale- and C. albicans-specific and non-specific humoral, lymphoproliferative and cytokine (IL-2, 4, 5 and IFNγ) responses in AD. Fifteen AD patients and seven healthy controls (HC) were included. Ficoll-isolated PBMC were stimulated by PHA and laboratory-generated extracts of P. ovale and C. albicans. Lymphocyte proliferation was measured by [sup 3]H-thymidine incorporation and cytokine production by sandwich-ELISAs. The antigen-specific IgG and IgE antibodies were analysed by ELISA and nitrocellulose RAST. Pityrosporum ovale- and C. albicans-specific IgE (both P < 0.001) and P. ovale-induced PBMC proliferation (P < 0.02) were elevated in AD. In general, the IL-4/IFNγ ratio induced by P. ovale was higher than that induced by C. albicans (P < 0.01). The PHA-induced IL-2 (P < 0.05) and IL-4 responses (P < 0.005), and the C. albicans-induced IL-5 response (P < 0.02) and IFNγ response (P < 0.01), were elevated in AD. A network of correlations was seen between serum total and the yeast-specific IgE, P. ovale-specific lymphoproliferation, PHA-induced IL-2, IL-4 and IL-5, and C. albicans-induced IL-5. The cytokine profiles found in this study support the role of TH[sub 0] or TH[sub 1] cells by the side of TH[sub 2] cells in the pathogenesis of atopic dermatitis. Pityrosporum ovale appears to be associated more with IL-4 responses and C. albicans with IFNγ responses. [ABSTRACT FROM AUTHOR]

Investigates the possible interaction of immature and mature antigen-presenting dendritic cells with the yeast Malassezia furfur and its allergenic components. Specific immunoglobulin reactivity of atopic dermatitis patients to the yeast cells; Confocal laser scanning microscopy of the yeast cells; Inhibition of pinocytosis.

Presents information on a study which investigated the allergen-induced cytokine responses of human peripheral blood mononuclear cells primed by tuberculin (PPD). Impact of Th2 cell production on allergic rhinitis; Role of mycobacteria in specific immunotherapy; Effects of PPD priming on mitchondrial RNA expression of Th2 cytokines.

Atopic dermatitis (AD) patients often demonstrate positive skin prick test results and serum IgE antibodies to a range of different yeasts. This has been thought to be due to cross‐reactivity. In this study, the cross‐reactivity of IgE and IgG antibodies between mannan and crude antigens of Pityrosporum ovale, Candida albicans, and Saccharomyces cerevisiae and crude antigens of Cryptococcus albidus and Rhodotorula rubra was examined by RAST and ELISA inhibition with two serum pools of AD patients. We found cross‐reacting IgE and IgG antibodies. In the IgE response, the main cross‐reacting pattern was the mannan region, although inhibition could be achieved also with crude antigens of C. albicans, S. cerevisiae, and, to some extent, C. albidus. P. ovale was the most potent inhibitor of IgE‐binding components, and against it the highest IgE antibody levels were detected in AD serum pools. In contrast, C. albicans was found to be the most important inducer of IgG antibodies, since the IgG level against P. ovale mannan in both AD serum pools was very low. Cross‐reacting antibodies were also seen in ELISA inhibition with both crude and mannan antigens, but since the IgG antibody level of P. ovale mannan in AD serum pools was low, further studies are needed to confirm the IgG results. [ABSTRACT FROM AUTHOR]

Examines the stability of Pityrosporum ovale allergens during storage. Induction of IgE antibody production in atopic dermatitis; Establishment of IgE response to P. ovale with skin-prick test.

Objective The aim of this study was to compare the IgE response to the most commonly farmed fur animals with that to domestic animals. Methods IgE-immunoblotting and RAST-inhibition analyses were performed using RAST-positive sera from fur workers sensitized to fur allergens and sera from patients sensitized to domestic animal allergens. Results The urine extracts of mink, blue fox, silver fox, racoon dog and fitchew contained more protein bands than the fur extracts did. Allergens with the same molecular weight were found in all of the fur and urine extracts. The most prominent allergenic bands had molecular weights of 62–67 kDa, 23–25 kDa and 18–19 kDa. With crossreacting sera the reciprocal RAST inhibition with all five animal extracts indicated common IgE-binding epitopes, probably common allergens (especially the 62–67 kDa bands). Urine and fur contain common allergens, since urine allergens strongly inhibited the IgE-binding to fur allergens. The IgE binding to allergenic bands of fur animal extracts was also observed in immmunoblotting when dog and cat RAST-positive sera were used, but not for cow RAST-positive sera. RAST inhibition of dog-positive sera with fur animal extracts and fur-positive sera with dog extract confirmed the crossreactivity of these IgE antibodies. No such inhibition was seen with cow extract. Conclusion The results of the RAST inhibition and immunoblotting suggest that fur animals have IgE binding epitopes or allergens in common with cat and dog — possibly albumin — but not with cow. [ABSTRACT FROM AUTHOR]

Background Pityrosporum ovale is a common saprophyte on the skin capable of inducing IgE antibody production in atopic dermatitis (AD) patients. Allergens of P. ovale have been examined in several studies, but consensus on them is lacking. Objective This study was carried out to obtain more information about the IgE antibody response against P. ovale, including mannan. Methods Sera from 64 AD patients and 10 healthy controls were analysed with immunoblotting and the nitrocellulose radio allergosorbent test (RAST) method specifically developed to detect antimannan P. ovale IgE antibodies. Results In immunoblotting a total of 39 different IgE stained protein bands were seen. A high molecular weight staining was also seen especially in patients who displayed elevated mannan P. ovale RAST values. The most commonly stained protein bands in immunoblotting were 9 and 96 kD bands with antibodies in 73 and 65'/f of AD patients who had been positive in commercial P. orbiculare RAST with total serum IgE less than 4000 kU/I. Mannan RAST appeared positive in 77% of them. Positive immunoblotting to either of these bands was seen in 90% and. if added with staining with the 20 kD band, in 100% of these AD patients. A combination of 9 kD IgE staining and mannan P, ovale RAST was positive in 92% of the patients and 96 kD and mannan P. ovale RAST in 85% of the patients. Conclusion It is evident that P. ovale has several allergens, the 9. 96 and 20 kD regions being the most important. According to our results mannan is also an important allegen of P. ovale. [ABSTRACT FROM AUTHOR]

Background Invasive candidiasis is a life-threatening complication problem in post- operative and immunocompromized patients, e.g. those treated by intensive care. Candida is frequently cultured from the mucous membranes of hospital patients and fungal cultures offer little diagnostic help. Other diagnostic methods, such as blood cultures, serology and diagnostic imaging techniques produce results too late and, if positive, low sensitivity. Objective To study the value of Candida-specific antibodies, especially those of IgE class, in diagnosing invasive Candida infection. Methods The immunoglobulins IgE, IgG and IgM responses to antigens of Candida albicans in the sera of 14 patients with culture, biopsy and/or autopsy proven postoperative invasive candidiasis and of 11 colonized and 19 non-colonized operated patients were studied by mannan radioallergosorbent test (PAST), mannan enzyme- linked immunosorbent assay (ELISA) and immunoblotting. Results Detection of IgE antibodies to C. albicans polysaccharide (mannan) and protein antigens proved specific and sensitive in diagnostics of invasive candidiasis after major abdominal surgery. IgE rose early in the course of the infection and the method made a clear distinction between invasive infection and mucous colonization. Immunoblotting for protein antibodies was most sensitive while nitrocellulose-RAST for mannan antibodies was most specific. The combined use of immunoblotting and RAST increased the sensitivity and the specificity. Determinations of anti-Candida IgG and IgM antibodies had low sensitivity and specificity. Conclusion Critically ill patients with invasive candidiasis develop IgE antibodies to Candida antigens probably because of disturbed TH1/TH2 responses. Determination of specific IgE antibodies can be used as a diagnostic aid in the early stage of invasive Candida infection. [ABSTRACT FROM AUTHOR]

Sera from 196 patients with a strong skin reactivity to Candida albicans and a history of atopic disease were used for C. albicans IgE-immunoblotting. The IgE-immunoblots were analysed by densitometry and integration ofthe densitogram peaks. A standardized reference disc system allowed parallel analysis of several different immunoblotting experiments. Altogether 105 patients had C albicans -specific IgE antibodies in immunoblotting. The IgE-response against C albicans was extremely polyspecific and was directed against 42 different bands. The most important IgE binding bands were 46 kDa and 27 kDa bands against which 44 (42%) and 29 (28%) patients, respectively, had IgE responses. A combination of IgE-immunoblotting and densitometry was found practical in analysis of large number of patient sera allowing a profound and accurate analysis and characterization of C. albicans allergen extract. This data evolved from a sufficiently large patient population can be utilized in further standardization of C. albicans extracts. [ABSTRACT FROM AUTHOR]

Ultrafiltered (> 1000 Da) samples of beer, aged red wine, young white wine, sparkling wine and extracts of fresh wheat bread and dried rye bread were analysed by skin-prick test (SPT), radioallergosorbent test (RAST) inhibition, sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting to find out if they contain Saccharomyces cerevisiae (S. cerevisiae. baker's yeast) allergens. Serum pool consisting of S. cerevisiae positive sera was used in the assays. The results were compared with freeze-dried reference S. cerevisiae and cereal antigens. The beer, bread, red wine and sparkling wine extracts elicited immediate reactions. However, no evident correlation with suspected symptoms was observed. White wine extract caused reactions in four out of six atopic dermatitis (AD) patients with symptoms, and in five out of seven symptom-free AD patients and in two of the 24 controls. In SDSPAGE, protein bands were found in wheat and rye bread extracts and beer. In IgE immunoblotting, however, no staining was seen with the S. cerevisiae positive sera suggesting that they were of cereal origin. In white wine and champagne extracts a nonspecific staining was seen in the region 20 kDa representing, e.g. lectin-like activity. No baker's yeast antigen could be detected in brewery and bakery products with IgE-immunoblotting even in the excessively concentrated extracts. The IgE mediated allergy to baker's yeast alone should thus not lead to denial of bakery, brewery and wine products. [ABSTRACT FROM AUTHOR]

The allergen extracts of wheat, rye, barley and oats flours were characterized by IgE-immunoblotting with serum samples from 40 adult patients; 35 patients with atopic dermatitis, one with rhinitis and four with urticaria. All these patients had been positive when skin-prick testing was carried out with one or more of the four flour extracts or displayed one or more positive cereal RAST results. Four non-atopic sera were used as negative controls. Acidic and neutral protein extracts of wheat, rye, barley and oats flours were processed for the immunoblotting experiments and 35 patients appeared positive in IgE-immunoblotting with wheat and rye, 32 with barley and 33 with oats. The IgE immunoblots showed polyspecific binding patterns; wheat exhibited 36 IgE stained bands, rye 35, barley 33 and oats 10. Eighteen of the IgE stained bands could be classified as intermediate allergens for wheat, 23 for rye and 15 for barley. The 66 kDa protein in oats was visualized by 28 out of 33 sera (84%), however, there was evident non-specific binding to this region and thus it may also represent lectin-like binding. The most frequent staining with wheat extract was seen in the 26 kDa protein region (15/35, 43%), with rye in the 40 kDa (16/35, 46%) and with barley in the 26 and 46 kDa protein bands (14/32, 44"A.). Simultaneous staining with wheat, rye and barley extracts were observed with 16 bands suggesting crossreactivity between these cereals. [ABSTRACT FROM AUTHOR]

A nitrocellulose-based radioallergosorbent test (RAST) was developed and used for the determination of IgE antibodies to Candida albicans mannan in patients with atopic dermatitis, asthma and allergic rhinitis. The results were expressed as mannan-RAST index values (an inter- and intra-assay coefficient for variation of 8.0–10.2%). The normal range for mannan-RAST index values was determined in 102 non-atopic adults. Fifty-three of 78 (67.9%) patients with atopic dermatitis showed elevated mannan-RAST index values with a significant correlation to the severity of the dermatitis (r = 0.33, P<001). Sixteen of 30 (53.3%) patients with asthma had a positive mannan-RAST index value; however, 12 of the 16 asthmatics (75%) who were positive also suffered from atopic dermatitis. Those who had allergic rhinitis but not atopic dermatitis showed a positive mannan-RAST index value in 12 of 32 (37.5%) cases. Nitrocellulose-RAST offered a sensitive method for the determination of polysaccharide-specific IgE antibodies in atopic diseases. The results show that high values are observed mainly in atopic dermatitis and less sensitization to C albicans occurs in respiratory allergy. [ABSTRACT FROM AUTHOR]

Fifty patients with farmer's lung disease (FL), 20 asymptomatic exposed farmers, and 18 healthy controls were investigated for total IgG and total IgA. as well as subclass antibody levels against Aspergillus umbrosus, the most common fungus to which FL patients in Finland have been exposed. Enzyme-linked immunosorbent assay (ELISA) revealed significantly elevated levels of specific total IgG and IgA antibodies in the sera of patients during the acute phase of the disease. Both IgG1 and IgG2 subclass antibodies and both IgA1 and IgA2 subclass antibodies were significantly associated with the disease. The antibody levels decreased during the first 3 months., when mould exposure was avoided and when an oral 6-week steroid treatment was administered. During the next 12 month follow-up, the mean antibody levels continued to decrease, even though re-exposure might have occurred. The results (with sensitivity 94% and specificity 95%) imply that in addition to the measurement of A. umbrosus-specific IgG, determination of both A. umbrosus-specific IgG1 and IgA might also be useful in the serodiagnosis of acute stage of FL. [ABSTRACT FROM AUTHOR]

An in vitro model was established to study the stability of baker's yeast (Saccharomyces cerevisiae) allergens in conditions simulating the gastrointestinal tract. The protocol consisted of 2 hr incubation under gastric conditions (pH 1.2, +37° C and gastric enzymes) and 2 hr incubation under duodenal conditions (pH 6.8, + 37° C and duodenal enzymes). These were studied together and separately, as well as under pure acidic conditions without gastric enzymes. The yeast extracts contained equal amounts of allergen and were analysed by IgE-immunoblotting. The acidic conditions had partly an enhancing and slightly degrading effect on the yeast allergens, whereas the gastric enzymes destroyed several allergens, including the important intermediate allergens of 31 and 45 kD. After treatment under both gastric and duodenal conditions most of the yeast allergens were destroyed, except mannan and a 10 kD protein component. The findings suggest that the allergen exposure caused by baker's yeast takes place mainly on the mucosal surfaces orally and oesophageally and through viable baker's yeast organisms that manage to pass the stomach and duodenum and possibly lead to intestinal growth of the organism. Patients with IgE production against the 10 kD allergen and mannan arc, however, moderately exposed to allergens consisting of soluble antigenic material only. [ABSTRACT FROM AUTHOR]

The role of sensitization and exposure to Candida albicans in atopic dermatitis (AD) was studied with skin-prick tests, yeast cultures and immunoblotting in 156 young adults with AD attending the Department of Dermatology, University of Turku, during 1983– 89. Eighteen patients with allergic rhinitis without eczema and 39 non-atopics were included as controls. Parameters associated with severe AD were simultaneous anti-C. albicans IgE and saprophytic C. albicans growth. A statistically significant correlation between C. albicans sensitization (specific IgE antibodies) and AD symptoms was observed only in patients with saprophytic C. albicans exposure. No correlation between C. albicans-specific IgE and AD severity was shown in patients without gastrointestinal growth. Furthermore, severe eczema was seldom seen in patients without saprophytic C. albicans growth. The most important IgE-binding components of C. albicans in immunoblotting were 27 and 46 kD proteins and mannan, a polysaccharide. IgG and IgA antibodies to C. albicans, mainly towards C. albicans mannan, were found in practically all 70 sera studied. These results suggest a continuous exposure and induction of IgE antibodies by C. albicans in AD patients. Severe phases of AD in colonized patients are associated with IgE synthesis against C. albicans. These findings suggest a role for C. albicans in the exacerbations of AD but the clarification of this subject needs double-blind placebo-controlled treatment trials. [ABSTRACT FROM AUTHOR]

Twenty-one different biotinylated lectins were used to recognize the carbohydrates of Aspergillus umbrosus, one of the most common microbes that patients with farmer's lung in Finland are exposed to. The glycoprotein fraction of A. umbrosus was bound especially well by Concanavalin A and consisted mainly of carbohydrates mannose and glucose. The carbohydrate fraction of A. umbrosus antigens were isolated from the crude extract of A. umbrosus with Con A- Sepharose affinity chromatography. Serum IgG antibodies to A. umbrosus mannoprotein fraction were determined in 57 patients with farmer's lung. 10 asymptomatic exposed farmers and 10 healthy controls by enzyme linked immunosorbent assay. Candida albicans and Saccharomyces cerevisiae mannan were used as controls. Patients had high levels, whereas asymptomatic exposed farmers showed moderate, and healthy controls low levels. of IgG antibodies to the A. umbrosus mannan/mannoprotein fraction. There were no significant differences in the mean antibody levels between the patients and controls against C. albicans or S. cerevisiae mannose fraction although in all groups more antibodies were detected against C. albicans than S. cerevisiae. [ABSTRACT FROM AUTHOR]

Stability of Candida albicans allergens was studied under various storage conditions. Lyophilized extract was reconstituted with human serum albumin (NSA) diluent, glycerol-free and in the presence of 10% or 50% glycerol and stored at various temperatures for different time periods. All extracts were tested at the same time with immunoblotting using C. albicans allergic patient sera and galactosidase-labelled anti-IgE. The highest number of detected allergens in the immunoblotting pattern was found in the presence of 50%) glycerol at +6°C. The most important allergen of C. albicans, the 46 kD protein allergen was stable up to 10 weeks at +6° C in the presence of 50% glycerol but thereafter began to lose its IgE-binding capacity. After 30 weeks more than 50% of the IgE binding had disappeared. The 27 kD protein, another important allergen, was also labile but retained the allergenicity better than the 46 kD one. The 29 kD protein allergen was stable at all storage conditions, except + 37°C tested even after one year. More than 6 months storage at +6°C or higher temperature is, however, unacceptable even in the presence of the 50% glycerol. These findings have particular importance in the diagnosis and treatment of allergic diseases. [ABSTRACT FROM AUTHOR]

Sera from 13 patients with positive Pityrosporum ovale skin prick test were analysed with IgE immunoblotting. Both P. ovale and Candida albicans antigens were used to reveal possible crossreactivity of these yeasts. Of the 13 sera, eight sera showed IgE binding to P. ovale and six sera to C. albicans. Simultaneous IgE-binding to both C. albicans and P. ovale was observed in five of these sera. The most common IgE-binding band pairs were, a 23 kD band of P. ovale and a 27 kD band of C. albicans, as one pair, and a 26 kD band and a 13 kD band, respectively, as another pair. In addition to these protein bands. simultaneous reactivity was observed to C. albicans mannan, a polysaccharide, and to a diffuse high molecular weight component of P. ovale, possibly also a polysaccharide. The five sera with simultaneous IgE-binding were analysed with RAST-inhibition, which revealed presence of crossreacting IgE-antibodies to P. ovale, and C. albicans in two of these sera. Crossreacting epitopes were suggested to be located in the mannan polysaccharide of C. albicans and high molecular weight diffuse stain of P. ovale, based on the immunoblotting and RAST-inhibition patterns of the studied sera. Crossreacting sera were pooled and used as an IgE probe in CRIE and Tandem-CRIE. These experiments revealed a fused precipitin line indicating presence of a common structure of P. ovale and C. albicans. This suggested crossreactivity may imply an interesting phenomenon in atopics who are sensitized by C. albicans growth in the gastrointestinal tract and get exposed by P. ovale growing on the skin. Further studies are thus needed to clarity, the clinical significance of these preliminary findings. [ABSTRACT FROM AUTHOR]

Immunoblotting and RAST were used to analyse IgE, IgA and IgG responses to antigens of Candida albicans. These were compared with the delayed skin response and C. albicans carriage in 40 atopic subjects. The majority of the atopic patients showed a strong IgG and IgA antibody response towards mannan, a carbohydrate, but only occasionally to proteins. Altogether 22 of the 40 patients showed specific IgE towards C. albicans by immunoblotting. The IgE response was mainly towards proteins, particularly to ones with molecular weights of 29 kD and 46 kD, and only in eight out of 22 IgE-positive subjects towards mannan. The IgG and IgA responses to mannan and the total IgE response towards C. albicans assessed by RAST showed an association with C. albicans carriage, whereas the delayed skin response showed an inverse relationship. The immunological parameters characteristic of C. albicans carriage were found to be C. albicans-specific, depressed delayed skin response and elevated IgE, IgA and IgG responses. This situation in the atopics presenting such parameters may favour simultaneous sensitization and exposure by colonization. The degree of sensitization may be sufficiently high to produce symptomatic allergy, such as asthma, in some individuals during occasional overgrowth of C. albicans, e.g. due to antibiotic therapy. [ABSTRACT FROM AUTHOR]

The carbohydrate-containing fraction of Candida albicans was isolated from the crude extract with ConA Sepharose affinity chromatography and studied by IgE-immunoblotting with individual and pooled sera from C.albicans-allergic subjects. In the ConA-bound fraction there was a diffuse IgE binding in the high molecular weight area which also gave a carbohydrate stain (PAS). A distinct band corresponding to a molecular weight of 70 kD bound specific IgE antibodies. This glycoprotein, presumably a mannoprotein, gave a weak carbohydrate staining and a strong protein staining. Further biochemical studies are needed to reveal the exact nature of the epitopes in the ConA-bound mannose-containing fraction of C.albicans. [ABSTRACT FROM AUTHOR]

Analysis of IgE, IgA and IgG antibodies directed against Candida albicans antigens in 28 asthmatic children was performed with immunoblotting after SDS-PAGE. Analysis with the purified cytoplasmic protein fraction revealed a major protein allergen with an MW of 46 kD. In addition to the major allergen, 15 other antigenic bands with molecular weights between 16 and 135 kD bound IgE. Ten of 13 anti-C. albicans IgE-positive children had IgE towards the 46 kD major allergen. None of the subjects in the study group or in the non-atopic controls had IgA or IgG antibodies towards this protein. Analysis of the crude surface extract showed that mannan, a carbohydrate, was an intermediate allergen contrary to being the major antigen in IgA and IgG antibody responses. [ABSTRACT FROM AUTHOR]

Water-soluble antigens of Candida albicans were sequentially extracted from intact and disrupted yeast cells grown on protein-free agar, and analysed on immunoblots after SDS-PAGE. Washing of the cells in saline before proper extraction resulted in loss of 47.2% of the total carbohydrate and 1.5% of the total protein. The protein fraction contained 14 antigenic bands when analysed with hyperimmune rabbit antisera. Four of these bound IgE when probed with a RAST-positive serum pool and beta-galactosidase-labelled anti-IgE. Extraction of the disrupted cells resulted in 15% of the total carbohydrate and 94% of the total protein. The cytoplasmic protein fraction showed 69 antigenic bands, 13 of which bound IgE. The carbohydrate fraction contained mannan, which was found in the washing solutions and in the surface extract as well as in the cytoplasmic extract. Allergens found in washing solutions were also present in cytoplasmic fraction. This study suggests that the rapid release of allergens from saprophytic C. albicans cells on mucous membranes of the body may cause continuous exposure and result in sensitization. [ABSTRACT FROM AUTHOR]

Yeast allergen extracts of Candida albicans, C. pseudotropicalis, C. krusei, C. parapsilosis, C. tropicalis, C. guilliermondi, C. humicola, C. norwegica, C. utilis, Cryptococcus albidus, Geotrichum candidum, Pityrosporon pachydermatis, P. ovale, Rhodotorula minuta, R. rubra, Saccharomyces cerevisiae, Torulopsis glabrata and Trichosporon cutaneum were investigated regarding their common allergenic properties. The enzyme immunoassay (EIA) using rabbit anti-Candida albicans antiserum showed remarkable immunological cross-reactivity only between the Candida species. However, there was a significant multiple sensitivity to the extracts of C. albicans, C. utilis, Cr. albidus, R. rubra and S. cerevisiae in skin prick testing in atopic patients, suggesting the possible presence of one or more common skin reactive allergens. [ABSTRACT FROM AUTHOR]

It is popular belief that mushrooms are capable of eliciting allergic symptoms. Studies on this subject are few and take no account of many of the important mushroom families, for example, in Scandinavia. The word mushroom refers mainly to macrofungi with edible fruiting bodies and other minor macrofungi. The word mushroom is used here in its widest sense. As the taxonomy is subject to continual review scientist have chosen to use the latest accepted latin name but mention in brackets the name used in the original article reviewed.

Background Farmer's lung (FL) is the most common form of extrinsic allergic alveolitis and the fungus Aspergillus umbrosus has frequently been associated with FL among Finnish farmers. IgG and IgA class antibodies against Aspergillus umbrosus are found in the serum of FL patients and healthy exposed farmers. So far the immunologically reactive subunits of Aspergillus umbrosus are unknown. Objectives The purpose of the study was to identify and localize the reactive subunits of Aspergillus umbrosus that are involved in the antigenic response in FL. Methods The distribution of the primary antibody on the outer wall layers of 4- and 11day-old fungi Aspergillus umbrosus was studied by pre-embedding immunolabelling with fluorescein isothiocyanate (FITC)- and gold-conjugated antibodies. The surface carbohydrates of the outer wall layers were studied using FITC-conjugated lectins. Results FITC-conjugated antibodies from FL patient sera reacted with spores (conidia), phialides and hyphae, whereas the fruiting body, cleistothecia, remained unstainable. Immunofluorescence labelling was confirmed by immunolabelling electron microscopy, and gold-conjugated IgG antibodies were found on the outer cell walls of spinulose conidia., and in a less degree on the outer cell walls of hyphae and phialides. Only single gold particles were found scattered on the surfaces of cleistothecia. The surface carbohydrates of Aspergillus umbrosus conidia, vesicles, phialides and hyphae had a high affinity to GlcNAcβ1- and Manαl-/Glcαl- specific lectins and only GlcNAβl-specific lectins also reacted with the outer cell walls of cleistothecia. Blocking of the antigenic sites of Aspergillus umbrosus by preincubating antigen-coated enzyme linked immunosorbent assay (ELISA) plates with lectins caused some inhibition, indicating that sites with the highest affinity for lectins were not always antigenic sites. Conclusion Gold- and FITC-conjugated antibodies were distributed mostly on the outer wall layers of conidia, hyphae and phialides of Aspergillus umbrosus and GlcNAcβ1-linked lectins had the strongest affinity to the outer wall layers. [ABSTRACT FROM AUTHOR]

Background Ingested cereals and inhaled cereal dusts can cause IgE-mediated allergy. Testing for these allergies yields variable, often undependable results, which could be due to the use of unsuitable or degraded testing materials. Objective We studied the effect of storage media, time and temperature on the stability of protein allergens extracted from wheat, rye, barley and oats flour. We also examined the effect of heat processing similar to that used when preparing foods on the stability of allergenic proteins present in wheat flour. Methods After storage experiments proteins in allergen extracts were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then visualized using immunoblotting or staining with Coomassie brilliant blue. After heat processing wheat flour proteins were extracted and their IgE-binding capacity was estimated. Results Extracted proteins from wheat, rye and barley flour appeared stable for at least 21 months when stored at 4°C in an aqueous solution containing 50% glycerol and 0.45% sodium chloride. In a medium with 0.4% phenol and 0.9% sodium chloride degradation of many proteins became apparent already after I month of storage. Oats extracts lost their IgE-binding capacity more rapidly and the maximal storage time appeared to be 10 months at 4°C in a solution containing 50% glycerol and 0.45% sodium chloride. The IgE-binding capacity of the allergenic proteins decreased as heat- processing temperature and heating time increased. No processing conditions entirely abolished IgE-binding. Conclusion Cereal allergen preparations can be stored either in lyophilized form or at 4°C in 50% glycerol. Unheated materials should be used also when testing for food allergy, as heating reduces allergenicity and thus sensitivity at testing. [ABSTRACT FROM AUTHOR]

Background Defective antibody response against bacterial polysaccharide antigens is known to be associated with recurrent pyogenic infections. The role of childhood allergy as a risk factor for repeated infections with capsulated micro-organisms has been controversial. Objective To compare the development of polysaccharide specific antibody responses in atopic and healthy infants and children. Methods The antibody responses against a common polysaccharide antigen, Candida albicans mannan, were studied longitudinally in IS atopic and 19 non-atopic children over the first 5 years of life. Determinations of IgA, IgG and IgM antibodies were carried out by enzyme-linked immunosorbent assay and IgE antibodies by nitrocellulose-based radioallergosorbent test. Results The polysaccharide specific antibody responses were similar in both groups, except that anti-mannan IgM levels were higher at 5 years in the atopic children (P <0.05, student's t-test). Conclusion Atopic children are not more susceptible to bacterial infections on the basis of poorer ability to produce antibodies against polysaccharide antigens. [ABSTRACT FROM AUTHOR]

Background: Hypersensitivity to cereals may occur via inhalation or ingestion. Although cereals are essential in the daily nutrition, only little information is available of the allergens causing symptoms in patients with atopic dermatitis (AD). Objective: The purpose of the present study was to analyse the IgE immune-response to various cereals and specific cereal fractions of wheat and oats in children with severe AD and correlate the results with challenge studies. Methods Skin-prick tests (SPT) with a NaCl suspension of wheat, oats, rice, corn, millet and buckwheat and the ethanol soluble gliadin fraction of wheat were performed to 34 wheat/oats challenge positive or negative children with AD. Simultaneously serum total IgE and specific IgE antibody radioallergosorbent test (RAST), levels to wheat, oats and gluten were determined. In addition serum samples of these 34 AD patients and five age matched controls were analysed with IgE immunoblotting using neutral and acidic protein extracts of wheat and oats. Results From the 34 AD children 33 were SPT positive with wheat and 18 with oats. Positive RAST to wheat and oats could be detected in 32 and 30 samples respectively. From the oral wheat challenge positive children 12/14 appeared positive with gliadin SPT and revealed positive RAST to gluten, but each of the wheat challenge negative were negative in SPT with gliadin. In immunoblotting using neutral and acidic fractions of cereals the IgE binding with sera of challenge positive children showed the most intensive staining, but no correlation was found between different staining patterns and the clinical wheat sensitivity. The 26, 38 and 69 kDa bands in wheat and the 46 and 66 kDa in oats could be classified as major IgE binding proteins of these cereals (> 50% of the sera were positive). SPT with rice, corn, millet or buckwheat was positive in 16/34 patients. Conclusion Intensive IgE staining to neutral/acidic soluble proteins in wheat and oats was seen, with major IgE binding to 26, 38 and 69 kDa proteins in wheat and 46 and 66 kDa in oats, but no specific IgE staining patterns correlating with clinical cereal sensitivity were found. The strong association between the positive oral wheat challenge and the positive SPT with the ethanol soluble gliadin suggests that also gliadin is an important allergen in wheat-allergic children with AD. The allergens in rice, corn, millet and buckwheat should be better studied before they can be recommended as alternatives for cereal allergic children. [ABSTRACT FROM AUTHOR]

This study was undertaken to analyze the differences in exposure and sensitization to five common environmental yeasts. The responses of IgG, IgA. and IgE to Candida albicans, C. utilis, Cryptococcus albidus. Rhodotorula rubra, and Saccharomyces cerevisiae and purified S. cerevisiae enolase were analyzed by immunoblotting (IgE-IB). and the cross-reactivity of their IgE-binding components by IgE-IB inhibition. Twenty atopic subjects, with asthma, allergic rhinitis, or atopic dermatitis were included. In skin prick tests (SPT). 12 of the patients showed simultaneous reactivity to at least two of the five yeasts, four reacted to one of the yeasts, and four had no responses. Antigens run in SDS-PAGE and transferred to nitrocellulose were probed with enzyme-labeled IgA-, IgG-, and IgE-specific antibodies. The IgE immunoblotting revealed most IgE-binding bands in C. albicans (11 bands) followed by C utilis (eight bands). S. cerevisiae (five bands), R. rubra (five bands), and Cr. albidus (four bands). Six of the IgE-binding bands of C. albicans and C. utilis shared molecular weight, and only two bands shared molecular weight with other yeasts. These were the 46-kDa band, shared by all five yeasts, and a 13-kDa band shared by four yeasts. Prominent IgE binding was seen to a 46-kDa band of C. albicans (seven patients), C. utilis (five patients), and S. cerevisiae (one patient) and to corresponding weak bands of Cr. albidus and R. rubra (one patient). The possible cross-reactivity of the 46-kDa band was analyzed by IgE-IB inhibition and densitometry, revealing clear C. albicans inhibition of C. utilis (80%) and enolase (98%) (autoinhibition 100%), The strongest IgG responses were seen against S. cerevisiae and C albicans. The responses were mainly against mannans of C. albicans and S. cerevisiae, suggesting that most of the exposure is to these yeasts. Yeasts with different types of exposure, from saprophytic growth on human mucous membranes to exposure by air and food, were shown to cross-react at the allergenic level, Atopic patients primarily sensitized by C. albicans and S. cerevisiae may develop allergic symptoms by exposure to other environmental yeasts due to cross-reacting IgE antibodies. [ABSTRACT FROM AUTHOR]

A standardized, controlled procedure for preparation of an in-house reference (IHR) preparation of an allergen extract of Candida albicans is described. The procedure, based on previous studies of allergens of C. albicans, is designed to yield a maximum of allergens in optimum extraction conditions and to provide a reference preparation for further extract production. The SDS-PAGE. igE-immunoblotting, and crossed radioimmunoelectrophoresis (CRIE) analyses showed that the procedure is reproducible with acceptable batch-to-batch variation. The variation in the content of the most important allergens, namely, proteins with molecular weights of 46, 29, and 27 kDa in the pooled final batches, is acceptable (coeff. of variation <15%), although in the intermediate batches of different strains, the coefficient of variation may occasionally exceed 20%. A comparison with other C. albicans allergen preparations used in our previous studies is also presented. The resulting extract can be used as a reference in further extract production and also in experimental in vitro and in vivo studies. [ABSTRACT FROM AUTHOR]

To determine the effects of anthropogenic pollution on water-soluble proteins and specifically allergens in birch (Betula pendula and B. pubescens) pollen, we analyzed extracts of pollen from the pollution gradient around a factory complex (emitting sulfur oxides and heavy metals) by sodium dodecyl sulfate (SDS) - polyacrylamide gel electrophoresis (PAGE) and IgE immunoblotting. In addition, tree density-associated shading of the tree habitat, and quantity and quality of proteins and allergens in pollen of the two birch species were studied. The two studied birch species gave identical allergen profiles even though their protein profiles differed. Distance from the factory did not affect the amount of birch pollen major allergen. Bet v 1 (17 kDa), or of two other strong allergens (23 and 36 kDa). Trees growing in shaded places had significantly stronger responses to Bet v 1 and to the 23-kDa allergen than trees growing in open or half-open environments. Thus, we propose that combined heavy metal and sulfur dioxide pollution does not have an important effect on birch pollen allergens. Instead, other factors, e.g., shading and soil properties of the tree habitat, as well as the genetic background of the tree, may have a stronger influence on the quantity and relative composition of allergens. [ABSTRACT FROM AUTHOR]

Clinical features, hypersensitivity mechanisms, and differential diagnosis of cereal allergy or intolerance were investigated in children with atopic dermatitis (AD). On oral provocation, 18 children exhibited a positive response to wheat, three to rye, one to barley, and one to oats. Cereal-induced symptoms were dermatologic, gastrointestinal, or oropharyngeal, and their onset after provocation was immediate (eight cases), delayed (14 cases), or both immediate and delayed (one case). A combination of type I allergy tests (prick test, RAST, and histamine-release test) detected all immediate reactors and &frac914; delayed reactors. Of the five subjects remaining negative in these tests, three were positive in the patch or lymphocyte-proliferation tests. Subjects with cereal allergy or intolerance frequently possessed IgE, IgA, and IgG antibodies against gliadin, but only one of these children was HLA-DR3-positive, and none had reticulin antibodies typical of celiac disease. Combining tests of immediate and delayed hypersensitivity can confirm allergy to cereals in a more reliable way. The coexistence of cereal allergy and celiac disease seems to be rare. [ABSTRACT FROM AUTHOR]

In this study, three immunologic tests, skin prick test, RAST, and basophil histamine-release test (BHRT), were compared by provocation in the diagnosis of occupational asthma and rhinitis. Twenty-three positive bronchial or nasal challenges were performed on 16 patients (six farmers, six bakery workers, and four food industry workers) and asthma or rhinitis was diagnosed as caused by cereal flour or grain, cow epithelium, storage mites, garlic, or soy dust. A control group consisted of 12 patients, of whom four (two bakery workers, one food industry worker, and one farmer) were challenge-negative, and the rest suffered from pollen allergy and seasonal rhinitis and were not challenged. The sensitivity and specificity of the prick test, RAST, BHRT, and a panel of them were as follows: 74 and 89%, 57% and 86%, 78 and 93%, and 91 and 71%, respectively. The overall concordance among these three type I allergy tests or between immunologic tests and challenge was relatively good. [ABSTRACT FROM AUTHOR]

The article examines the allergens of Ficus benjamina indoor plant. About three to four percent of all atopics exhibit skin reaction to the plant. Ficus benjamina sap extracts have been characterized in an immunoblotting study. The sap extract serves as the best source of the plant allergens. Allergens in the sap are species-specific.

Examines extrinsic allergic alveolitis from pine sawdust. Increase in the total amount of cells in the bronchoalveolar fluid; Collection of sawdust from pine, spruce, birch, beech and apple; Methods in preparing antigen extracts.

Background: Food allergen immunotherapy (FA-AIT) practice is known to vary globally. This project aims to identify and characterize European centres performing FA-AIT., Methods: An EAACI task force conducted an online survey to gather relevant information regarding FA-AIT practice and setting-specific resources after reviewing the published literature and congress abstracts throughout Europe., Results: We identified 102 FA-AIT centres in 18 countries; only Spain (n = 39) and France (n = 16) had ≥10 such centres. Overall, most facilities were hospital-based (77.5%), publicly funded (80.4%) and delivered FA-AIT as routine clinical care (80.4%). On average, departments had 3 allergists/paediatric allergists and 2 nurses. Surveyed centres had provided FA-AIT for a median of 9 years [1-24] to a median of 105 [5-2415] patients. The estimated total number of treated patients was 24875, of whom 41.3% received AIT for milk, 34.2% egg, 12.8% peanut and 11.7% other foods. Anaphylaxis to AIT doses requiring over 4-6 h of observation was reported by 70.6% of centres, ICU admissions by 10.8% and eosinophilic esophagitis by 45.1%. Quality of life and sustained unresponsiveness were evaluated in 20.6% and 54.9% of centres, respectively. The main contraindications for food AIT were severe asthma (57%-63%), eosinophilic esophagitis (56%-48%) and age below 5 years (47%-41%)., Conclusions: In Europe, FA-AIT is provided mostly in clinical practice. Significant variation is seen in the number of centres per country, facility characteristics and inclusion/exclusion criteria, and in certain aspects of protocols. Potential inequality in access to AIT has been identified as well as the need for education and guidance for treatment standardization., (© 2021 European Academy of Allergy and Clinical Immunology and John Wiley & Sons Ltd.)