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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]

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]

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]

Local reactions and pertussis toxin specific immunoglobulin E antibodies (PT-IgE) were investigated in healthy children following primary and booster immunization with a combined diphtheria tetanus acellular pertussis vaccine (DTPa) including pertussis toxin, filamentous haemagglutinin and pertactin. A primary series of DTPa was administered to 150 infants, and 104 of them received a booster dose of DTPa combined with inactivated polio vaccine at 2 years of age. PT-IgE was measured in serum samples from 72 children using a modified nitrocellulose RAST. Primary immunization was associated with low incidence of local reactions (1%-5%). After the booster dose 21% of children had a local reaction >/=20 mm. Local reactions after the booster dose tended to be more common in children who had experienced reaction at primary immunization. PT-IgE was detected in 18% and 86% of children following primary and booster vaccinations, respectively. Allergic and non-allergic children did not differ in PT-IgE responses. After primary immunization, elevated PT-IgE levels were found more often in children with a family history of allergy than in those without known allergy in the family. Children with local reactions had significantly higher pre- and post-booster PT-IgE levels and median post-booster pertactin IgG and diphtheria-IgG levels than children without local reactions. Conclusion Acellular pertussis immunization induces IgE antibodies to pertussis toxin, especially after booster vaccination. The higher median pre- and post-booster levels of pertussis toxin specific immunoglobulin E and post-booster levels of IgG to pertactin and diphtheria in children with local side-effects reflect a multifactorial immunological mechanism of such reactions. [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]

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]

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]

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]

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 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]

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]