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Background: Specific IgE (sIgE) antibodies to both bee and wasp venom can be due to a sensitivity to both insect venoms or due to cross-reactive carbohydrate determinants (CCDs)., Objective: Investigating whether a basophil activation test (BAT) with both venoms as well as with bromelain and horseradish peroxidase (HRP) or recombinant allergen-based IgE testing can improve the diagnostic procedure., Methods: Twenty-two Hymenoptera-venom allergic patients with sIgE antibodies to both bee and wasp venom were studied. sIgE antibodies to MUXF3 CCD, bromelain, HRP, rApi m 1, and rVes v 5 were determined, and a BAT (Flow2 CAST) with venom extracts, bromelain, and HRP was performed. Further recombinant allergen-based IgE testing was done by using an ELISA, if required. The reactivity of basophils was calculated from the insect venom concentration at half-maximum stimulation., Results: Double positivity/double negativity/single positivity to rApi m 1 and rVes v 5 was seen in 12/1/9 patients. Further recombinant allergen-based IgE testing in the last ones revealed positive results to the other venom in all cases except one. BAT was double positive/double negative/single positive in 6/2/14 patients. Four patients with negative results in sIgE antibodies to CCDs had positive results in BAT. BAT with bromelain/HRP showed a sensitivity of 50%/81% and a specificity of 91%/90%., Conclusion: Component-resolved IgE testing elucidates the pattern of double positivity, showing a majority of true double sensitizations independent of CCD sensitization. BAT seems to add more information about the culprit insect even if the true clinical relevance of BAT is not completely determined because of ethical limitations on diagnostic sting challenges. BAT with HRP is a good method to determine sensitivity to CCDs., (Copyright © 2012 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.)

Background: The identification of the disease-causing insect in venom allergy is often difficult., Objective: To establish recombinant allergen-based IgE tests to diagnose bee and yellow jacket wasp allergy., Methods: Sera from patients with bee and/or wasp allergy (n = 43) and patients with pollen allergy with false-positive IgE serology to venom extracts were tested for IgE reactivity in allergen extract-based tests or with purified allergens, including nonglycosylated Escherichia coli-expressed recombinant (r) Api m 1, rApi m 2, rVes v 5, and insect cell-expressed, glycosylated rApi m 2 as well as 2 natural plant glycoproteins (Phl p 4, bromelain)., Results: The patients with venom allergy could be diagnosed with a combination of E coli-expressed rApi m 1, rApi m 2, and rVes v 5 whereas patients with pollen allergy remained negative. For a group of 29 patients for whom the sensitizing venom could not be identified with natural allergen extracts, testing with nonglycosylated allergens allowed identification of the sensitizing venom. Recombinant nonglycosylated allergens also allowed definition of the sensitizing venom for those 14 patients who had reacted either with bee or wasp venom extracts. By IgE inhibition studies, it is shown that glycosylated Api m 2 contains carbohydrate epitopes that cross-react with natural Api m 1, Ves v 2, natural Phl p 4, and bromelain, thus identifying cross-reactive structures responsible for serologic false-positive test results or double-positivity to bee and wasp extracts., Conclusion: Nonglycosylated recombinant bee and wasp venom allergens allow the identification of patients with bee and wasp allergy and should facilitate accurate prescription of venom immunotherapy., (Copyright (c) 2010 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.)

IgE, mast cells, basophils, and eosinophils are essential components of allergic inflammation. Antigen-specific IgE production, with subsequent fixation of IgE to FcepsilonRI receptors on mast cells and basophils, is central to the initiation and propagation of immediate hypersensitivity reactions. Mast cells, basophils, and eosinophils are central effector cells in allergic inflammation, as well as in innate and adaptive immunity. This review highlights what is known about these components and their roles in disease pathogenesis., (Copyright 2010 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.)

Clinical laboratory analyses aid in the diagnosis and management of human allergic (IgE-dependent) diseases. Diagnosis of immediate-type hypersensitivity begins with a thorough clinical history and physical examination. Once symptoms compatible with an allergic disorder have been identified, a skin test, blood test, or both for allergen-specific IgE antibodies provide confirmation of sensitization, which strengthens the diagnosis. Skin testing provides a biologically relevant immediate-type hypersensitivity response with resultant wheal-and-flare reactions within 15 minutes of allergen application. Allergen-specific IgE antibody in serum is quantified by using 3 laboratory-based autoanalyzers (ImmunoCAP, Immulite, and HYTEC-288) and novel microarray and lateral-flow immunoassays. Technologic advances in serologic allergen-specific IgE measurements have involved increased automation, with enhanced reproducibility, greater quantification, lower analytic sensitivity, and component-supplemented extract-based allergen use. In vivo provocation tests involving inhalation, ingestion, or injection of allergens serve to clarify discordant history and skin- or blood-based measures of sensitization. Other diagnostic allergy laboratory analyses include total and free serum IgE measurement, precipitating IgG antibodies specific for organic dusts, mast cell tryptase, and indicator allergen analyses to assess indoor environments to promote patient-targeted allergen avoidance programs. A critique is provided on the predictive utility of serologic measures of specific IgE for food allergy and asthma. Reasons for the lack of clinical utility for food-specific IgG/IgG4 measurements in allergy diagnosis are examined. When the specific IgE measures are inconsistent with the clinical history, they should be confirmed by means of repeat and alternative method analysis. Ultimately, the patient's clinical history remains the principal arbiter that determines the final diagnosis of allergic disease., (Copyright 2010 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.)

Background: Maternal history of asthma or atopy is among the most consistently reported risk factors for asthma and atopy in children, yet the molecular basis for this observation remains unclear. Mitochondria are inherited exclusively through the maternal line, raising the possibility that sequence variation in the mitochondrial genome contributes to the pathogenesis of asthma and atopy., Objective: We set out to determine whether common European mitochondrial haplogroups are associated with asthma-related atopic phenotypes., Methods: We studied 654 self-reported white children age 5 to 12 years with mild to moderate asthma participating in the Childhood Asthma Management Program. Eight haplogroup-tagging polymorphisms were genotyped with TaqMan probe hybridization assays, and mitochondrial haplogroup tests of association with asthma-related and atopy-related phenotypes were performed with haplo.stats., Results: We found significant evidence of mitochondrial haplogroup association with total serum IgE levels (global significance, P = .04), with carriers of European haplogroup U (frequency 11%) having higher total serum IgE levels (median level, 684 IU/L) compared with noncarriers (389 IU/L; P = .001). Haplogroup U carriers also had trends of greater skin prick test reactivity (P = .03) and higher frequency of atopic dermatitis (P = .07), although global haplogroup tests for these later 2 phenotypes were not significant at an alpha level of 0.05., Conclusion: These data are the first to suggest that common mitochondrial haplogroups influence the atopic diathesis., Clinical Implications: These findings may provide a molecular explanation for the prominent influence of maternal history of atopy on the development of atopy in offspring.

Background: Contradictory results have been found when analyzing the IgE cross-reactivity among pyroglyphid mites and storage mites, as well as the role of these as true sensitizing agents., Objective: We sought to study the prevalence of sensitization to Glycyphagus domesticus in patients naturally exposed to this mite together with the more ubiquitous Dermatophagoides pteronyssinus and the IgE cross-reactivity between them., Methods: Mite species present in house dust samples of consecutive patients with mite allergy from Huelva (southwest Spain) were determined by means of light microscopy. Clinical sensitization was assessed by using skin prick and conjunctival provocation tests. Cross-reactivity at the IgE level was studied by using serum IgE determination and inhibition of RAST, IgE immunoblotting, and mite group 2 immunoassays., Results: After D pteronyssinus, which is present in about 95% of house dust samples, G domesticus was the most important mite and present in about 50% of the samples. Tyrophagus putrescentiae and Lepidoglyphus destructor were detected in third and fourth place, respectively. About half of the patients with G domesticus at home were sensitized to this mite. A low IgE cross-reactivity was observed between D pteronyssinus and G domesticus, but an important IgE cross-reactivity was detected among glycyphagid mites at the level of group 2 allergens., Conclusion: Glycyphagid mites can act as primary sensitizing agents independently of pyroglyphid mites in a subset of patients naturally exposed to them., Clinical Implications: The inclusion of glycyphagid mite extracts in the diagnostic battery in areas with adequate mite growing conditions is important to ensure proper diagnosis.

Background: Epidemiologic data on allergic rhinitis and asthma are frequently based on self-reported symptoms., Objective: This cross-sectional study examined the relationship between self-reported symptoms and histories of allergic rhinitis or asthma and a marker of allergic sensitization, allergen-specific IgE., Methods: We surveyed 702 pregnant women in Michigan. Blood samples were analyzed for specific IgE to 9 allergens: dust mites (Dermatophagoides farinae and Dermatophagoides pteronyssinus), cat, dog, cockroach, ragweed, timothy grass, egg, and Alternaria alternata. Seratopy was defined as a specific IgE greater than or equal to 0.35 kU/L to any allergen., Results: Seroatopy was found in 66.7% of those with hay fever symptoms, 68.3% with a physician's diagnosis of asthma, and 72.1% of those with both conditions. These results differed significantly from asymptomatic subjects, where 49.8% of patients without hay fever and 50.4% without asthma were seroatopic. Race and education did not modify the relationships. Symptoms related to specific exposures were modest predictors of positive specific IgE to related allergens (positive predictive values from 26.5% to 50.3%)., Conclusion: Self-reported symptoms of allergic rhinitis or asthma were significantly associated with allergic sensitization, but the odds ratios were of relatively low magnitude for this historical information to be considered evidence of current allergic sensitization. A 66% to 68% probability existed that those with symptoms of allergic rhinitis or asthma would have a positive specific IgE test., Clinical Implications: Self-reported histories of hay fever or asthma alone are only modest predictors of allergic sensitization. When knowledge of allergic sensitization is important, information beyond self-reported symptoms is necessary.

Background: Allergens from cat are among the most potent elicitors of allergic disease. Four cat allergens have been identified; however, evidence indicates the existence of additional allergens., Objective: In this study, we evaluated IgE sensitization to IgA from cat., Methods: Sera from cat-sensitized patients (n = 81) were analyzed for IgE antibodies to purified cat IgA in the Pharmacia CAP System. Indirect ELISA was performed with cat IgA, cat IgM, and deglycosylated cat IgA. Competitive inhibition ELISA was performed with cat IgA, cat IgM, calf intestine alkaline phosphatase (CIP), and cat serum albumin on solid phase bound cat IgA. IgE reactivity was also evaluated on membrane blotted cat IgA., Results: Thirty-eight percent (31/81) of the cat-sensitized sera were ImmunoCAP-positive to cat IgA. Indirect ELISA demonstrated a high correlation between IgE reactivity to cat IgA and cat IgM (r = 0.94; P < .001). Very low responses were observed to deglycosylated IgA. Strong inhibition of cat IgA was observed in all sera after preincubation with cat IgA and cat IgM. Inhibition was also observed in most sera after preincubation with CIP. Immunoblotting demonstrated that the IgE reactivity was mainly directed to the heavy chain of IgA., Conclusion: This study has revealed a new allergen, cat IgA, containing a novel group of cross-reactive epitopes depending on carbohydrates also present on IgM and partially on CIP., Clinical Implications: This new group of cross-reactive carbohydrate IgE epitopes should be taken into consideration when diagnosing patients with suspected animal allergy.

Background: Individuals with pollen allergy often have IgE against plant-derived foods. This can be due to cross-reactive IgE against Bet v 1 and homologues, profilins, and/or cross-reactive carbohydrate determinants., Objective: The aim of this study was to correlate sensitization to Bet v 1 and profilin with individual recognition patterns to plant foods and clinical relevance., Methods: Fifty-two patients with pollen allergy and IgE against at least one plant-derived food were included in the study. Adverse reactions to plant-derived foods were documented by using standardized interviews. Skin prick tests were performed for pollen (grass, birch, and mugwort) and 14 plant-derived foods. In addition, recombinant (r) Bet v 1 and rBet v 2 (profilin) were tested intracutaneously. Specific IgE against the abovementioned allergens were determined by means of RAST. Cross-reactivity was studied by means of RAST inhibition., Results: Eighty-five percent of patients were sensitized to Bet v 1, and 71% were sensitized to profilin. Profilin was associated with a higher number of positive RAST results to plant-derived foods than Bet v 1. In contrast, Bet v 1 was associated with more positive skin prick test responses and more food-related symptoms. Sensitization to Bet v 1 was associated with IgE against apple, hazelnut, and peach, whereas sensitization to profilin was associated with positive RAST results to all investigated plant-derived foods except apple, peach, and melon., Conclusions: IgE antibodies against Bet v 1 have a more limited spectrum of cross-reactivity than those against profilin, but they frequently give rise to clinically relevant cross-reactivities to food. In analogy to anticarbohydrate IgE, cross-reactive IgE against food profilins have no or very limited clinical relevance.

Background: It has been proposed that Trichophyton infection is associated with atopy and allergy., Objectives: Our purpose was (1) to confirm whether atopy predisposes to chronic dermatophytosis and (2) to investigate whether Trichophyton infection induces atopic disease., Methods: Patients attending dermatology clinics and suspected of having dermatomycosis underwent in a prospective manner fungal culture and Trichophyton and inhalant skin tests, and blood serum was collected for total IgE and Trichophyton radioallergosorbent testing. Personal and family history of atopic diseases was also investigated., Results: According to mycologic culture, atopic history, and inhalant skin test results, patients were classified into 4 groups: (1) atopy plus mycosis (n = 28), (2) atopy (n = 26), (3) mycosis (n = 35), and (4) no atopy, no mycosis (n = 33). Patients with active mycosis (groups 1 and 3) demonstrated significantly increased positivity of Trichophyton skin tests compared with patients without fungal infection (groups 2 and 4), regardless of their atopic status, whereas atopic patients (those in groups 1 and 2) had significantly increased levels of total serum IgE compared with nonatopic subjects. Trichophytosis was not more prevalent in atopic than in nonatopic subjects, and atopic diseases were not more frequent in culture-positive than in culture-negative patients., Conclusions: Our results indicate that Trichophyton -specific IgE is observed in patients with trichophytosis regardless of atopy.

Serum IgE levels were followed throughout the first year of life in a sample of 34 infants from atopic and nonatopic families. Their parents' serum IgE levels were also measured. Neonatal levels ranged from 0 to 10 U/ml. Half of the study group, which included a high proportion of infants from atopic families, maintained serum IgE levels below 10 U/ml until 1 yr of age. In the remainder, the serum IgE exceeded 10 U/ml before 1 yr of age, but the time of increase and the rate of increase varied from infant to infant; the sooner the IgE started to increase, the higher was the serum IgE level at 1 yr of age. The relationship of serum IgE level to the development of atopic disease and to other parameters was analyzed. Elevation of serum IgE level at or before 1 yr of age was highly correlated with atopic disease in the first 2 yr of life. The elevation preceded the manifestation of atopy.

Total serum IgE levels were determined in 136 newborns and their mothers and in 54 of their fathers, using the paper radioimmunosorbent test (PRIST) technique. IgE specific antibodies for house dust (Dermatophagoides pteronyssinus), orchard grass, timothy grass, and cow's milk were measured with the radioallergosorbent test (RAST). One hundred thirty-three RAST assays were negative in newborns, and in three cases RAST for cow's milk was positive. Cord blood IgE ranged from 0 to 5.5 IU/ml (mean 0.32 +/- 0.54 IU/ml); levels were significantly (p less than 0.05) higher when maternal IgE was over 100 IU/ml and when mothers had received progesterone therapy during the pregnancy. Salbutamol administration or tobacco smoking during pregnancy did not influence newborn IgE. A clinical follow-up study was conducted in 83 infants for 9 mo. Nine infants developed definite atopic disease, and possible allergic diseases were noted in eight other infants. The IgE level at birth appeared to be more predictive for the development of allergy in infancy than the family history.

Highly atopic infants often form IgE antibodies toward multiple food protein in the first 2 years of life. They begin producing IgE antibody to inhalant allergens between the first and second year of life. We hypothesized that highly atopic children would be at significant risk of sensitization to peanut. We defined high atopy as serum IgE greater than or equal to 10 times 1 SD from normal plus multiple positive RASTs. In this study we have characterized the immunologic status of 141 patients by measuring total serum IgE and specific IgE to several allergens, including peanut. These data demonstrated that, independent of clinical history, a positive RAST to peanut was more common in the highly atopic category compared to the low atopy category. Significantly more patients who were highly atopic and had a positive peanut RAST had a positive RAST for egg or milk compared to low atopic patients. More significantly, 33 of the patients had never knowingly received peanut, yet 21 (63.6%) had a positive RAST for peanut, whereas seven (21.2%) had a peanut antibody in the highest RAST category. All these seven patients were considered highly atopic according to the definition above, and three were younger than 2 years of age. These results suggest that highly atopic infants are at special risk for sensitization to peanut, even when they have never received peanut, and that characterization of immunologic sensitization to milk, egg, and peanut will identify the highly atopic infant.(ABSTRACT TRUNCATED AT 250 WORDS)

We describe in this article a fully automated, universal assay for serum IgE after pepsin digestion of the sample and subsequent assay of the released Fc" epsilon fragment by particle counting immunoassay (PACIA). The sensitivity and the range of the assay were easily modulated by changing the concentration of dextran in the reaction medium. In the application to cord serum, the sensitivity reached 0.1 IU/ml in 30 min incubation time and with a throughput of 50 analyses per hour. Within-day and between-day coefficients of variation did not exceed 7.6% for IgE levels covering a wide range of the standard curve. Dependable accuracy was demonstrated by linearity tests, analytical recoveries (89% to 112%), and correlation with PRIST on 48 samples from children ages 1 to 14 yr (y = 0.97x + 5.92; r = 0.987). Minor discrepancies between the two methods were attributed to a slight serum effect in PRIST. PACIA applied to 348 cord serum samples demonstrated a statistically significant influence of sex and race on the cord IgE level. In European neonates boys had significantly (p = 0.019) higher geometrical mean cord IgE levels (n = 142; 0.46 IU/ml; range less than 0.10 to 30 IU/ml) than girls (n = 146; 0.33 IU/ml; range less than 0.10 to 8.0 IU/ml), which was attributed to a predominance of boys (chi 2 = 4.29; p less than 0.05) having more often elevated cord IgE (less than 1.20 IU/ml) than girls. Neonates of African-Asian origin had significantly (p less than 0.00005) higher cord IgE levels (n = 60; 1.05 IU/ml; range less than 0.10 to 125.0 IU/ml) than European neonates (n = 288; 0.39 IU/ml; range less than 0.10 to 30.0 IU/ml).