Early indoor aeroallergen exposure is not associated with development of sensitization or allergic rhinitis in high-risk children.

RATIONALE: Allergen exposure is associated with the development of allergic sensitization in childhood as reflected by global variations in sensitization patterns. However, there is little evidence to support a direct association. OBJECTIVES: To investigate the association between perinatal aeroallergen exposure and sensitization and rhinitis to such allergens later in childhood. METHODS: Allergic sensitization to cat, dog, and house dust mites was diagnosed longitudinally using skin prick tests and specific IgE measurements at ½, 1½, 4, 6, and 13 years in 399 children from the Copenhagen Prospective Study on Asthma in Childhood2000 birth cohort. Rhinitis was diagnosed at 7 and 13 years. Allergen exposure was defined as dog or cat in the home during the 3rd trimester of pregnancy or the first year of life and as allergen levels of dog, cat, and house dust mite in bed dust samples at 1 year. Associations between exposure and outcomes were analyzed by logistic regression and stratified for eczema status and test method (skin prick test and specific IgE). RESULTS: We found no association between dog or cat exposure in perinatal life and sensitization or rhinitis during childhood. Similarly, there was no association between levels of allergens in bed dust samples and sensitization or rhinitis during childhood. CONCLUSION: Perinatal indoor aeroallergen exposure does not seem to affect development of allergic sensitization or rhinitis during childhood questioning the relevance of allergen avoidance as a preventive measure. Other factors such as timing of allergen exposure or other environmental adjuvants may contribute in a more complex pathway to sensitization.

Inhibitors of mammalian target of rapamycin downregulate MYCN protein expression and inhibit neuroblastoma growth in vitro and in vivo.

Mammalian target of rapamycin (mTOR) has been shown to play an important function in cell proliferation, metabolism and tumorigenesis, and proteins that regulate signaling through mTOR are frequently altered in human cancers. In this study we investigated the phosphorylation status of key proteins in the PI3K/AKT/mTOR pathway and the effects of the mTOR inhibitors rapamycin and CCI-779 on neuroblastoma tumorigenesis. Significant expression of activated AKT and mTOR were detected in all primary neuroblastoma tissue samples investigated, but not in non-malignant adrenal medullas. mTOR inhibitors showed antiproliferative effects on neuroblastoma cells in vitro. Neuroblastoma cell lines expressing high levels of MYCN were significantly more sensitive to mTOR inhibitors compared to cell lines expressing low MYCN levels. Established neuroblastoma tumors treated with mTOR inhibitors in vivo showed increased apoptosis, decreased proliferation and inhibition of angiogenesis. Importantly, mTOR inhibitors induced downregulation of vascular endothelial growth factor A (VEGF-A) secretion, cyclin D1 and MYCN protein expression in vitro and in vivo. Our data suggest that mTOR inhibitors have therapeutic efficacy on aggressive MYCN amplified neuroblastomas.

Respiratory symptoms, asthma and allergen levels in schools--comparison between Korea and Sweden.

UNLABELLED: We studied reports on respiratory symptoms, asthma and atopic sensitisation in relation to allergen contamination in Korean schools and compared with data from a previous Swedish study performed in eight primary schools. Korean pupils (n = 2365) in 12 primary schools first completed a questionnaire. Then airborne and settled dust were collected from 34 classrooms and analyzed for allergens by ELISA. In both countries, boys reported more symptoms. The prevalence of wheeze was similar, while daytime [odds ratio (OR) = 14.0, 95% confidence interval (CI) = 9.0-21.9] and nocturnal breathlessness (OR = 3.1, 95% CI = 1.5-6.4) were much higher among Korean students. In Korean schools, dog allergen (Can f 1) was the most common followed by mite allergen (Der f 1), while cat (Fel d 1), dog, and horse allergen (Equ cx) were abundant in Sweden. Moreover, CO(2) levels were high in most Korean schools (range 907-4113 ppm). There was an association between allergen levels in dust and air samples, and number of pet-keepers in the classrooms. In conclusion, allergen contamination in Korean schools may be an important public issue. PRACTICAL IMPLICATIONS: This study showed that furry pet allergen contamination was common in both Korean and Swedish schools. In addition, house dust-mite (Der f 1) allergen contamination was common in Korean schools, probably because of transport of allergen from other environments. Transfer should therefore be minimized. Korean schools had high CO(2) levels and the concept of mechanical ventilation should be introduced. Measurement of airborne allergen levels is quite new and seems to be a more convenient and correct way to monitor allergen exposure in classrooms.

Indoor molds, bacteria, microbial volatile organic compounds and plasticizers in schools--associations with asthma and respiratory symptoms in pupils.

UNLABELLED: We investigated asthma and atopy in relation to microbial and plasticizer exposure. Pupils in eight primary schools in Uppsala (Sweden) answered a questionnaire, 1014 (68%) participated. Totally, 7.7% reported doctor-diagnosed asthma, 5.9% current asthma, and 12.2% allergy to pollen/pets. Wheeze was reported by 7.8%, 4.5% reported daytime breathlessness, and 2.0% nocturnal breathlessness. Measurements were performed in 23 classrooms (May-June), 74% had <1000 ppm CO(2) indoors. None had visible mold growth or dampness. Mean total microbial volatile organic compound (MVOC) concentration was 423 ng/m(3) indoors and 123 ng/m(3) outdoors. Indoor concentration of TMPD-MIB (2,2,4-trimethyl-1,3-pentanediol monoisobutyrate, Texanol) and TMPD-DIB (2,2,4-trimethyl-1,3-pentanediol diisobutyrate, TXIB), two common plasticizers, were 0.89 and 1.64 microg/m(3), respectively. MVOC and plasticizer concentration were correlated (r = 0.5; P < 0.01). Mold concentration was 360 cfu/m(3) indoors and 980 cfu/m(3) outdoors. At higher indoor concentrations of total MVOC, nocturnal breathlessness (P < 0.01) and doctor-diagnosed asthma (P < 0.05) were more common. Moreover, there were positive associations between nocturnal breathlessness and 3-methylfuran (P < 0.01), 3-methyl-1-butanol (P < 0.05), dimethyldisulfide (P < 0.01), 2-heptanone (P < 0.01), 1-octen-3-ol (P < 0.05), 3-octanone (P < 0.05), TMPD-MIB (P < 0.05), and TMPD-DIB (P < 0.01). TMPD-DIB was positively associated with wheeze (P < 0.05), daytime breathlessness (P < 0.05), doctor-diagnosed asthma (P < 0.05), and current asthma (P < 0.05). In conclusion, exposure to MVOC and plasticizers at school may be a risk factor for asthmatic symptoms in children. PRACTICAL IMPLICATIONS: Despite generally good ventilation and lack of visible signs of mold growth, we found an association between respiratory symptoms and indoor MVOC concentration. In addition, we found associations between asthmatic symptoms and two common plasticizers. The highest levels of MVOC, TMPD-MIB, and TMPD-DIB were found in two new buildings, suggesting that material emissions should be better controlled. As MVOC and plasticizers concentrations were positively correlated, while indoor viable molds and bacteria were negatively correlated, it is unclear if indoor MVOC is an indicator of microbial exposure. Further studies focusing on health effects of chemical emissions from indoor plastic materials, including PVC-floor coatings, are needed.

A comparative study of asthma, pollen, cat and dog allergy among pupils and allergen levels in schools in Taiyuan city, China, and Uppsala, Sweden.

We compared the school environment, asthma and allergy in 10 schools in Taiyuan, China, with eight schools in Uppsala, Sweden. In total 2193 pupils (mean age 13 years) participated. Chinese pupils had more respiratory symptoms, particularly daytime breathlessness after exercise (29.8% vs. 7.1%; P < 0.001), while cat allergy (1.2% vs. 6.6%; P < 0.001) and dog allergy (1.3% vs. 4.0%; P < 0.01) was less common. Cumulative incidence of asthma (1.8% vs. 9.5%; P < 0.001) and doctor's diagnosed asthma (1.2% vs. 9.0%; P < 0.001) were less common in China, indicating an under-diagnosis of asthma. Chinese classrooms were colder (mean 14.7 vs. 21.4 degrees C), more humid (mean 42% vs. 31% RH) and had higher CO2-levels (mean 2211 vs. 761 ppm). Levels of cat (Fel d1), dog (Can f1) allergens were low in settled dust from China (< 200 ng/g dust), but high in airborne dust on Petri-dishes (GM 16.8 ng/m2/day for Fel d1 and 17.7 for Can f1). The Swedish settled dust contained cat, dog and horse allergens in high levels (median 1300 ng/g, 1650 ng/g, 1250 U/g dust, respectively). In conclusion, there were large differences in the school environment, and in respiratory symptom and allergy. Allergen measurements in settled dust only may largely underestimate the classroom exposure. Practical Implications There is a need to improve the school environment, both in China and Sweden. The Swedish schools contained high levels of cat, dog and horse allergens and more amounts of open shelves and textiles that can accumulate dust and allergens. The air measurements indicated that Chinese schools may contain significant amounts of cat and dog allergen, and analysis of settled dust only may not reflect the true allergen exposure. Since the Chinese schools had no mechanical ventilation, they could not fulfill the ventilation standard in winter, and hence there is a need for improving the ventilation. The great discrepancy between respiratory symptoms and reports on asthma, and the high prevalence of attacks of breathlessness without wheeze, may have implication for future questionnaire studies on asthma in China.

Current asthma and respiratory symptoms among pupils in relation to dietary factors and allergens in the school environment.

UNLABELLED: The aim was to study asthma and allergy in relation to diet and the school environment. Pupils (5-14 years) in eight schools received a questionnaire, 1014 participated (68%). Settled dust was collected on ALK-filters and analyzed for allergens from cat (Fel d 1), dog (Can f 1), horse (Equ cx), house dust mites (Der p 1, Der f 1), and cockroach (Bla g 1) by ELISA. In total, 6.8% reported cat allergy, 4.8% dog allergy, 7.7% doctor's diagnosed asthma and 5.9% current asthma, and 7.8% reported wheeze. Current asthma was less common among those consuming more fresh milk (P < 0.05) and fish (P < 0.01). Poly-unsaturated fatty acids was associated with more wheeze (P < 0.05), olive oil was associated with less doctors' diagnosed asthma (P < 0.05). Totally, 74% of the classrooms had mean CO(2) <1000 ppm. The median concentration per gram dust was 860 ng/g Fel d 1, 750 ng/g Can f 1 and 954 U/g Equ cx. Horse allergen was associated with more wheeze (P < 0.05), daytime breathlessness (P < 0.05), current asthma (P < 0.05) and atopic sensitization (P < 0.05). Dog allergen was associated with wheeze (P < 0.05) and daytime breathlessness (P < 0.05). The associations between allergens and respiratory symptoms were more pronounced among those consuming margarine, not consuming butter, and with a low intake of milk. In conclusion, cat, dog and horse allergens in schools could be a risk factor for asthma and atopic sensitization, and dietary factors may interact with the allergen exposure. PRACTICAL IMPLICATIONS: Previous school studies performed by us in mid-Sweden, showed that most classrooms did not fulfill the ventilation standards. In this study, most of the classrooms fulfilled the ventilation standard, but despite that had widespread allergen contamination. Most previous studies have focused on cat allergen, but our study has shown that also dog and horse allergens can be risk factors for asthma and allergy in schools. As allergens are transported from other environments, mainly the home environment, the main prevention should be to minimize transfer of allergens. This could be achieved by reducing contacts with furry pets and horses, or using different clothes at home and at school (e.g. school uniforms). Increased cleaning in the schools may reduce allergen levels, but the efficiency of this measure must be evaluated in further intervention studies. Finally, our study supports the view that dietary habits among pupils should not be neglected and interaction between dietary factors and indoor allergen exposure needs to be further investigated.

An ELISA for recombinant Lepidoglyphus destructor, Lep d 2, and the monitoring of exposure to dust mite allergens in farming households.

BACKGROUND: Exposure to indoor allergens, such as dust mites, has been recognized as a risk factor for sensitization and symptoms. OBJECTIVE: To develop a two-site ELISA for the determination of Lep d 2 in the reservoir, to measure dust mite allergen exposure (Lep d 2, Der p 1, Der f 1 and Der 2) in farm households, and to investigate whether exposure to these allergens is associated with sensitization, asthma and rhinoconjunctivitis. METHODS: Monoclonal antibodies to recombinant (r)Lep d 2 were produced with standard hybridoma technique. Dust samples from 393 households were analysed for allergen content by two-site ELISA methods. RESULTS: A two-site Lep d 2 ELISA was developed with a detection limit of 0.09 microg/g. The assay was highly reproducible and levels of Lep d 2 showed a strong correlation with the number of Lepidoglyphus mites (r(s): 0.7; P = 0.0002). Lep d 2 was detected in 20% of the homes; levels ranged from 0.09 to 1.7 microg/g of dust. Der p 1 was recorded in 59% of the samples, ranging from 0.055 to 139 microg/g, and Der f 1 and Der 2 in 40% and 50% of the samples, ranging from 0.055 to 24.5 microg/g and 24.3 microg/g, respectively. Dermatophagoides allergens were significantly higher in mattresses than in carpets (P < 0.0001), but this difference was not observed with Lep d 2. A strong relationship between immunoglobulin (Ig)E to rLep d 2 and asthma (OR = 10.4) and rhinoconjunctivitis (OR = 7.5) was seen. Furthermore, sensitization to D. pteronyssinus was significantly associated with asthma (OR: 13.7) and rhinoconjunctivitis (OR: 5.7). CONCLUSION: When assessing mite allergen exposure in rural homes, not only the Der p 1, Der f 1 and Der 2 allergens, but also the Lep d 2 allergen should be taken into consideration.

Development of new IgE specificities to allergenic components in birch pollen extract during specific immunotherapy studied with immunoblotting and Pharmacia CAP System.

BACKGROUND: New IgE sensitizations to proteins in allergen extracts have been shown to occur during allergen-specific immunotherapy (IT). However little is known about the kinetics of the changes in antibody reactivities. METHODS: Twenty-four allergic children and adults were treated with birch pollen rush IT (RIT). Fifteen matched patients served as allergic controls. Sera were obtained at regular intervals for up to three years and analyzed with immunoblotting and Pharmacia CAP System with recombinant (r) birch pollen allergens (rBet v 1, rBet v 2, and rBet v 4). RESULTS: All birch-allergic patients had specific IgE to the major birch pollen allergen Bet v 1, but only three had IgE to rBet v 2 and/or rBet v 4 at the beginning of the study. New IgE sensitizations developed in 65% of the birch RIT-treated patients when studied by immunoblotting. Twenty-nine percent of the patients developed new sensitizations to rBet v 2 and/or rBet v 4 during RIT as measured by Pharmacia CAP System. Generally, new specific IgE reactivities occurred after at least one year of RIT, and only at low levels (< 1 kUA/l). CONCLUSIONS: Sensitization to additional allergenic pollen components frequently occurs during prolonged birch RIT. However, the IgE levels are low and the clinical relevance is not known.

T cell responses to recombinant isoforms, synthetic peptides and a mutant variant of Lep d 2, a major allergen from the dust mite Lepidoglyphus destructor.

BACKGROUND: The dust mite Lepidoglyphus destructor is an important cause of allergic reactions to dust, especially in farming environments. Two isoforms, recombinant (r)Lep d 2.01 and rLep d 2.02, of the major allergen Lep d 2, have previously been expressed as recombinant proteins. These isoforms differ 10.4% at the amino acid level. Furthermore, a mutant form of Lep d 2.01 (rLep d 2.6Cys) with a highly reduced IgE reactivity, has also been produced. OBJECTIVE: To investigate the T cell responses to the recombinant isoforms of Lep d 2, the Lep d 2.6Cys mutant and peptides of Lep d 2, in allergic and non-allergic individuals. METHODS: Peripheral blood mononuclear cells from 18 allergic and 16 non-allergic individuals were stimulated with the different antigens and the proliferative responses were measured. The cytokine production (interleukin (IL)-4, IL-5 and interferon (IFN)-gamma) were measured by ELISA. RESULTS: Higher T cell proliferation was measured to isoform 01 than to 02 in 28/34 subjects. The responses to rLep d 2.6Cys were lower than to isoform 01 in most subjects, but higher than to Lep d 2.02. Two immuno-dominant peptides, corresponding to amino acid residue 11-25 and 61-75 were identified. The atopic subjects produced significantly lower IFN-gamma in response to Lep d 2.01 as compared to the non-atopics. CONCLUSIONS: There was a significant difference in T cell response between the two isoforms of rLep d 2. The hypoallergenic mutant rLep d 2.6Cys was able to evoke a T cell response with a magnitude which is between the two isoforms. Amino acid residue 11-25 and 61-75 are the most frequently recognized parts of Lep d 2 and are likely to contain the immuno-dominant T cell epitopes.

Pollen-specific rush immunotherapy: clinical efficacy and effects on antibody concentrations.

BACKGROUND: Studies of rush immunotherapy (RIT) with standardized extracts for the treatment of seasonal pollen allergy are few, especially for birch-pollen RIT. OBJECTIVE: The study was performed to investigate the efficacy of RIT with standardized birch- or timothy-pollen extracts. Further, the serum antibody levels were evaluated for correlation with clinical efficacy. METHODS: This open, longitudinal study included 30 allergic patients treated with RIT and 16 allergic patients serving as a control group. The therapy was continued for 3 years and blood samples were collected at regular intervals for antibody measurements using the Pharmacia CAP System. RESULTS: The RIT was generally well tolerated. An increase in the total and specific IgE concentrations during the early months of RIT was observed, followed by decreased levels. Specific IgG and IgG4 increased continuously for 2 years. The symptom and medication scores were significantly decreased, compared with preRIT, at both the first and third pollen seasons after the start of RIT treatment (P < .0001 and P < .001, respectively). The clinical improvement during RIT was significantly greater compared with the control group (P < .05). The decreased medication and the symptom improvement during the third year of RIT correlated with the relative decrease in specific IgE (rs = .52, P < .05) and with the specific IgG4 level before the start of RIT (rs= -.68, P < .01), respectively. CONCLUSIONS: Our study indicates that RIT with standardized birch- or timothypollen extracts is clinically effective and safe. Measurements of specific antibody levels during treatment may be helpful in monitoring RIT.

IgA cross-reactivity between a nuclear autoantigen and wheat proteins suggests molecular mimicry as a possible pathomechanism in celiac disease.

Celiac disease patients display IgA antibody reactivity to wheat as well as to human proteins. We used serum IgA from celiac patients and, for control purposes, from patients with Crohn's disease, ulcerative colitis and from healthy individuals to identify celiac disease-specific IgA autoantigens in nitrocellulose-blotted extracts from various human cell types (epithelial, endothelial, intestinal cells, fibroblasts). The pattern, recognition intensity and time course of IgA autoreactivity was monitored using serial serum samples obtained from celiac children before and under gluten-free diet. By immunoblot inhibition and subcellular (cytosolic, nuclear) cell fractionation we identified a 55 kDa nuclear autoantigen expressed in intestinal, endothelial cells and in fibroblasts which was recognized by IgA antibodies of approximately half of the celiac disease patients and cross-reacted with wheat proteins. IgA reactivity to the 55 kDa autoantigen disappeared during gluten-free diet and was inhibited after pre-absorption of sera with wheat proteins but not with tissue transglutaminase, previously reported as the unique celiac disease-specific autoantigen. In conclusion, we defined a novel 55 kDa celiac disease-specific nuclear IgA autoantigen which shares epitopes with wheat proteins and which is different from tissue transglutaminase and calreticulin. Although the newly defined autoantigen was recognized much less frequently than tissue transglutaminase, our data suggest molecular mimicry between wheat and human proteins as a possible pathomechanism for the induction and/or maintenance of mucosal tissue damage in celiac disease.

A new method for collecting airborne allergens.

BACKGROUND: Measurement of airborne allergens has hitherto been done with the use of fixed-location pumps or personal air samplers. Our objective was to find out whether ionizers could be good tools for collecting airborne allergens. As a model we have used cat allergen (Fel d l). We have compared Fel d l levels collected by the ionizer at different time periods, as well as comparing Fel d l levels obtained with the ionizer with those of low- and high-volume pumps. METHODS: Dust samples from floors and air samples collected with ionizers and pumps, obtained in 31 homes with cat, 23 homes without cats, and 28 day-care centres, were analysed for cat allergen content (Fel d I) by ELISA. RESULTS: Fel d l was present in the reservoir in all homes with cats, ranging from 660 to 375,000 ng/g (GM 75,000) and in the air collected by the ionizer from 2.0 to 204 ng/24 h (GM 19.3). The allergen in homes without cat varied from < 55 to 1,800 ng/g (GM 166). Corresponding levels in air were found in two of these homes (2.3 and 7.3 ng/24 h). There was a correlation between the number of cats and the amount of airborne cat allergen (r: 0.47; P < 0.05). The levels in day-care centres were < 55 to 3,070 ng/g in dust (GM 360) and < 1.1 to 7.9 ng/24 h in the air (GM 1.6). We obtained a moderately strong correlation between air and dust samples in homes with cats (rs: 0.64; P< 0.001) and in day-care centres (rs: 0.49; P<0.05). We found that a collection period of 24 h is preferable for the ionizer. The intrahome reliability coefficient was nearly two times higher for the ionizer (r: 0.69) than the pump (r: 0.39). CONCLUSIONS: The ionizer seems to be a good tool for monitoring the environment. It is easy to use and silent and does not disturb the airflow in the room.

Cytokine production by peripheral blood mononuclear cells following birch-pollen immunotherapy.

We studied the Th2/Th1 balance by short-term stimulation of peripheral blood mononuclear cells (PBMC) isolated during the pollen season from seven allergic patients treated with conventional birch-pollen immunotherapy (IT) for 18 months, eight matched allergic control patients and 10 non-atopic individuals. The PBMC were cultured for 7 days with birch-pollen extract (BPE) or tetanus toxoid (TT), and then restimulated with PHA and PMA to induce high IL-5, IL-10 and IFN-gamma production. The serum levels of birch-pollen-specific IgG and IgG4 were significantly elevated after IT treatment. The proliferative response to BPE was significantly enhanced in the allergic control group, but not in the IT-treated group, compared to the non-atopic group (P<0.05). Birch-pollen-specific IL-5 production was significantly enhanced in both the IT-treated group and the allergic control group (P<0.01-0. 05). Furthermore, both the IT-treated group and the allergic control group had a cytokine profile to BPE significantly more Th2 polarized (high IL-5/IFN-gamma ratio) than to TT (P<0.05 and P<0.01, respectively). No differences in IL-10 production between the three study groups were observed. The Th2/Th1 balance in vitro correlated with the serum concentrations of birch-pollen-specific IgE (r=0.60, P<0.05), and in the IT-treated group, also with the IgG and IgG4 levels (r=0.79, P<0.05 and r=0.86, P<0.05, respectively). We conclude that conventional birch-pollen IT does not lead to changes in the cytokine profile of the circulating pool of allergen-specific T cells during birch-pollen season. However, induction of peripheral T-cell tolerance and increased production of specific IgG and IgG4 might be part of the mechanisms of IT.

Changes in cytokine production in vitro during the early phase of birch-pollen immunotherapy.

The kinetics of the immunological mechanisms during allergen-specific immunotherapy (IT) has not been thoroughly evaluated. In this investigation we study the changes in T-cell responses during the early phase of IT. Ten patients (IT group) with birch-pollen allergy were treated with conventional IT. Blood samples were collected at regular intervals for specific immunoglobulin (Ig)E measurements and preparation of peripheral blood mononuclear cells (PBMC). Seven allergic control patients (AC group) were included during the subsequent birch-pollen season. The PBMC were stimulated with birch-pollen extract or tetanus toxoid (TT) and mitogens. After a short decrease, probably owing to seasonal variation, the birch-pollen-specific proliferation and the interleukin (IL)-4, IL-5, and IL-10 production significantly increased when reaching the maintenance dose and during the subsequent pollen season. The increase in IL-4 correlated with a temporary increased serum level of birch-pollen-specific IgE. Interestingly, also the TT-specific response was affected by IT, resulting in weaker, but in time similar, changes in proliferation and cytokine production as in the birch-pollen-specific response. We speculate that the early phase of IT might lead to systemic changes in the capacity of Th2-like cytokine production, and that the early increase in allergen-specific IgE is a consequence of enhanced IL-4 production.

Antibody reactivity against human and guinea pig tissue transglutaminase in children with celiac disease.

BACKGROUND: Highly discriminatory markers for celiac disease are needed to identify children with early mucosal lesions. The purposes of this study were to evaluate the clinical potential of circulating anti-tissue transglutaminase (tTG) immunoglobulin (Ig)A antibodies in the diagnosis of childhood celiac disease and to investigate the extent of autoreactivity of these antibodies. METHODS: Included in this retrospective study were samples from 22 children with biopsy-verified celiac disease, 23 control subjects with disease, and 22 healthy control subjects without any known gastrointestinal or inflammatory disorders. An enzyme-linked immunosorbent assay (ELISA) was used to measure the serum levels of IgA antibodies specific for human and guinea pig tTGs. All samples were also analyzed for antibodies to gliadin and endomysium (EMA). RESULTS: The concentrations of IgA specific for human and guinea pig tTGs correlated with the small intestinal villous structure and the serum levels of IgA EMA. The tTG ELISAs exhibited a high specificity and sensitivity for detection of untreated celiac disease. The human erythrocyte IgA tTG ELISA had the highest sensitivity (100%) and a specificity of 98%. The IgA EMA method had a sensitivity of 95% and the highest specificity (100%) of all tests. CONCLUSIONS: Our results provide additional support to the concept that anti-tTG IgA antibodies can be used as a highly discriminatory serologic marker for celiac disease and that measurements of these autoreactive antibodies may in the future be used as an alternative to the EMA test.

Study of the Th1/Th2 balance, including IL-10 production, in cultures of peripheral blood mononuclear cells from birch-pollen-allergic patients.

BACKGROUND: Excessive production of interleukin (IL)-4, IL-5, IL-10, and IL-13 is thought to be important in the development of allergy and asthma. The objective of this investigation was to study Th1/Th2-like cytokine profiles in vitro in seven patients allergic to birch pollen and six nonallergic controls during the birch-pollen season. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated and cultured with birch-pollen extract (BPE) or tetanus toxoid (TT) for 7 days, harvested, and restimulated with the mitogens phytohemagglutinin (PHA) and phorbol 12-myristate 13-acetate (PMA) for 24 h. Cytokine production was determined by ELISA, and logarithmic cytokine ratios were compared between the two groups and between the antigens. RESULTS: In the allergic group, the cultures prestimulated with BPE had a more Th2-like cytokine response than the TT-prestimulated cultures; i.e., lower IFN-gamma and higher IL-10 production (P<0.05), as well as higher IL-5/IFN-gamma and IL-13/ IFN-gamma ratios (P<0.05). There were also significantly higher IL-4/IFN-gamma (P<0.005) and IL-5/IFN-gamma (P<0.05) ratios in BPE-stimulated cultures in the allergic group than in the control group. The IL-4 and IL-13 production in vitro correlated with the specific serum IgE levels. CONCLUSIONS: BPE stimulation induces a Th2-like cytokine response by PBMC isolated during the pollen season from birch-pollen-allergic patients, indicating a Th2-type immune response to birch pollen in vivo.

Sensitization to airborne and food allergens in Reykjavík (Iceland) and Uppsala (Sweden) - a comparative study.

BACKGROUND: The aim of this study was to compare the prevalence of atopic sensitization and possible risk factors for allergies in two ethnically similar but geographically widely separated urban populations. METHODS: Data from two centers of the European Community Respiratory Health Survey, Reykjavik, Iceland, and Uppsala, Sweden, were utilized. This included a structured interview, skin prick tests, and blood samples for total and specific IgE for common aeroallergens. Additional measurements of specific IgE antibodies to common food antigens were performed. Furthermore, data on social environment, lifestyle, air pollution, and meteorologic variables were compared. RESULTS: Skin prick tests were done on 540 individuals in Reykjavik and 527 in Uppsala. The overall prevalence of at least one positive prick test was 20.5% in Reykjavik and 34.2% in Uppsala (P<0.001). Total and specific IgE were measured in serum from 521 subjects in Reykjavik and 472 in Uppsala. The geometric mean value for total IgE was significantly lower in Reykjavik (13.4 kU/l) than in Uppsala (24.7 kU/l) (P<0.001). Similarly, the overall prevalence of at least one specific IgE to airborne allergens was 23.6% in Reykjavik and 32.3% in Uppsala (P<0.01). Specific IgE to a food panel (fx5) was measured in 502 subjects in Reykjavik, and 434 in Uppsala. In Reykjavik, 20 individuals (4.0%) were positive to one or more of the allergens in the food panel compared to 27 (6.0%) in Uppsala. When the single allergens present in the food panel were measured, altogether 16 positive reactions were found in Reykjavik compared to 47 in Uppsala (P<0.05). CONCLUSIONS: The prevalence of sensitization to both airborne and food allergens was lower in Reykjavik than in Uppsala. The difference may be due to environmental and/or dietary differences or to some yet undefined factor.

IgE binding capacity of synthetic and recombinant peptides of the major storage mite (Lepidoglyphus destructor) allergen, Lep d 2.

BACKGROUND: Lepidoglyphus destructor is an important non-pyroglyphid mite species in Europe and a dominant allergen in farming environments. The major allergen of L. destructor, Lep d 2, is a protein of 13.2 kD that is recognised by about 90% of sera RAST positive to this mite species. METHODS: The cDNA of two isoallergens of the Lep d 2 has previously been sequenced and the protein expressed in different protein expression systems. In order to map the B-cell epitopes, the full length protein and the truncated forms of the protein have been expressed in Escherichia coli as glutathione-S-transferase (GST) fusion proteins. Recombinant Lep d 2 fragments and synthetic overlapping 15 mer peptides spanning Lep d 2 were probed with sera from patients allergic to storage mite. RESULTS: The full-length (125 amino acids) GST fusion protein reacted strongly with patient IgE in Western blots and dot blots. Synthetic peptides failed to react with IgE antibodies from mite-allergic patients and the truncated fusion proteins displayed weak IgE-binding capacity. CONCLUSION: We conclude that there are no dominant linear IgE-binding epitopes in Lep d 2. Recombinant or synthetic Lep d 2 fragments may, however, be further evaluated as hypoallergenic candidate molecules for specific immunotherapy.

IL-8 and the activation of eosinophils and neutrophils following nasal allergen challenge.

BACKGROUND: A growing body of evidence suggests that proinflammatory cytokines play a role in allergic inflammation by attracting and activating inflammatory cells. In this study, we have investigated the relationship between interleukin-8 (IL-8) in nasal lavage fluid and the local activation of eosinophils and neutrophils following nasal allergen challenge of allergic patients. METHODS: Nasal challenges were performed with grass pollen extract in 14 allergic patients and 5 nonallergic controls. Nasal lavage fluid was collected repeatedly for 10 h, and the levels of eosinophil cationic protein (ECP) and myeloperoxidase (MPO) were used as markers of eosinophil and neutrophil activation, respectively. The levels of these molecules were compared with that of IL-8 in nasal lavage fluid. RESULTS: Allergen challenge of allergic patients produced a significant late-phase increase in the levels of ECP and MPO. Furthermore, the level of MPO showed a highly significant correlation with the level of IL-8 in lavage fluid (r = 0.8, p< 0.0001), whereas there was no significant relationship between the levels of ECP and IL-8. CONCLUSION: Interestingly, our findings suggest that both eosinophils and neutrophils are activated following nasal allergen challenge. In addition, our results are consistent with the hypothesis that IL-8 acts as a chemoattractant/activator of neutrophils during the late phase of the allergic inflammation. In contrast, we were not able to demonstrate any significant relationship between the level of IL-8 in lavage fluid and the activation of eosinophils.

Allergen-specific increase in interleukin (IL)-4 and IL-5 secretion from peripheral blood mononuclear cells during birch-pollen immunotherapy.

The mechanisms behind the effects of immunotherapy (IT) with birch-pollen extract are largely unknown. In this pilot study, we measured the cytokine secretion in vitro from peripheral blood mononuclear cells (PBMC) obtained from birch-pollen-allergic patients undergoing IT treatment (n = 4) or placebo administration (n = 4), collected before treatment, 1 and 4 weeks after start of treatment, and during and just after the pollen season (12-14 weeks after start of treatment). The PBMC were stimulated with birch-pollen extract in vitro for 7 days, followed by restimulation with the mitogens phytohemagglutinin (PHA) and phorbol 12-myristate 13-acetate (PMA) for 24 h, to enhance the production of cytokines. The supernatants were analyzed with ELISA and radioimmunoassay for interferon-gamma (IFN-gamma), interleukin (IL)-4, and IL-5. In the therapy group, we noted an increased secretion of IL-4 and IL-5 from PBMC collected at 4 weeks after the start of treatment (IL-4: 29 +/- 21 pg/ml [day 0] to 374 +/- 448 pg/ml [week 4], mean +/- SD; IL-5: 95 +/- 48 pg/ml to 1147 +/- 697 pg/ml). No increase was seen in the placebo group. During the pollen season, we noted a trend toward increased IL-4 and IL-5 secretion in both groups. We conclude that the temporary increase in serum IgE observed in many IT studies may be a consequence of increased IL-4 production due to the allergen exposure.