Commercial Allergen Extracts Research Articles

IgE Cross-Reactivity between Humulus japonicus and Humulus lupulus.

PurposeJapanese hop (Humulus japonicus) is a major cause of weed pollinosis in East Asia. However, supplies of commercial allergen extract from this plant have not met clinical demand. The pollen of common hop (Humulus lupulus), a closely related species, may provide an alternative source if there is strong IgE cross-reactivity between these two species. We aimed to compare the IgE cross-reactivity and allergenicity of common hop and Japanese hop pollen.Materials and MethodsCross-reactivity was measured by inhibition ELISA. One- and two-dimensional (2D) gel analyses combined with IgE immunoblotting and mass spectrometry [liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS)] were performed to detect IgE-reactive pollen components.ResultsUp to 16.7% of IgE reactivity to Japanese hop was inhibited by common hop. A 12-kDa protein component of Japanese hop pollen that showed the most potent IgE reaction was absent from common hop. Six IgE-reactive components from Japanese hop were detected by 2D gel electrophoresis and LC-ESI-MS/MS, but showed low Mascot scores, preventing positive identification.ConclusionNo significant IgE cross-reaction was observed for Japanese and common hop pollen allergens. Development of allergy diagnostic and immunotherapeutic reagents based on Japanese hop pollen are urgently needed.

Original Article. Sensitization to Aeroallergens in Pediatric Patients with Allergic Rhinitis and Asthma

Summary Asthma and allergic rhinitis are common in childhood. Establishing sensitization to aeroallergens is crucial to effectively prevent exacerbation of these respiratory allergic diseases. The study aimed to evaluate sensitization to the most common aeroallergens in children with asthma and allergic rhinitis.We establishedacorrelation between patients sensitized to indoor, outdoor and simultaneously to both allergens. The study population consisted of 276 patients (168 boys and 108 girls) ages 4 to 16 years with asthma (A) and allergic rhinitis (AR). Skin prick tests were performed with 21 commercial allergen extracts: pollens, mites, epithelia and insects, and molds. We found that 217 patients were sensitized to at least one aeroallergen: 117 patients hadapositive result to mites, 92 to pollens, 72 to epithelia and insects, and 63 - to mold allergens. Dermatophagoides pteronyssinus was the most prevalent aeroallergen. Sensitized only to indoor allergens were 104 patients, 60 - only to outdoor allergens, and 53 were sensitized to both. Mites were the most frequent aeroallergens in children with Aand AR. Lately there has been foundasignificant increase in rates of sensitization to mold allergens, especially to Alternaria alternata. Our study has confirmed the dominant role of indoor allergens in children with respiratory allergic diseases.

An alternative inhibition method for determining cross-reactive allergens

Inhibition assays are an useful tool to identify the allergen of primary sensitization of cross-reactive allergens. Classical ELISA-based inhibition assays are limited by both the availability of commercial standardized allergen extracts and the experience and knowledge needed for making home-made extracts. Moreover the direct comparison of the inhibition ELISAs outcomes between different laboratories is difficult because of different sources of used allergen extracts and a number of methodological variations. Therefore, we propose a novel ImmunoCap (Phadia, Thermofisher Scientific) based immunoinhibition method with the use of commercially available Caps as the allergen source. The novel ImmunoCap based immunoinhibition method was developed and tested with sera from patients with a well-known cross-reactive sensitization for fig (Ficus carica) and ficus (Ficus benjamina). Results were compared with a classically applied inhibition method, i.e. addition of homemade allergen extract to patient serum. The amount of allergens (fig and ficus extracts) needed to reach a similar degree of inhibition was comparable for both inhibition methods. The ImmunoCap based inhibition assay, in addition to classical inhibition methods, is a valuable tool as the ImmunoCap analyzer and commercial allergens (Caps) are more widely available which makes the outcomes of inhibition tests comparable between different laboratories. Furthermore, in the ImmunoCap inhibition method the same protein source is used for both the inhibition of sIgE and sIgE measurement, which might be even more relevant when multiple cross-reactive allergens are tested.

Measurement of Major Allergen Mus m 1 in Commercial Mouse Allergen Extracts and Mouse Urine

Measurement of Major Allergen Mus m 1 in Commercial Mouse Allergen Extracts and Mouse Urine

Multiplex Assay for Protein Profiling and Potency Measurement of German Cockroach Allergen Extracts.

BackgroundGerman cockroach (GCr) allergens induce IgE responses and may cause asthma. Commercial GCr allergen extracts are variable and existing assays may not be appropriate for determining extract composition and potency.ObjectiveOur aim was to develop a multiplex antibody/bead-based assay for assessment of GCr allergen extracts.MethodsSingle chain fragment variable (scFv) antibodies against GCr were obtained by screening libraries derived from naïve human lymphocytes and hyperimmunized chicken splenocytes and bone marrow. Selected clones were sequenced and characterized by immunoblotting. Eighteen scFv antibodies (17 chicken, 1 human) coupled to polystyrene beads were used in this suspension assay; binding of targeted GCr allergens to antibody-coated beads was detected using rabbit antisera against GCr, and against specific allergens rBla g 1, rBla g 2, and rBla g 4. The assay was tested for specificity, accuracy, and precision. Extracts were also compared by IgE competition ELISA.ResultsChicken scFv’s generated eight different binding patterns to GCr proteins from 14 to 150 kDa molecular weight. Human scFv’s recognized a 100 kDa GCr protein. The multiplex assay was found to be specific and reproducible with intra-assay coefficient of variation (CV) of 2.64% and inter-assay CV of 10.0%. Overall potencies of various GCr extracts were calculated using mean logEC50s for eight selected scFvs. Overall potency measures were also analyzed by assessing the contributions to potency of each target.ConclusionsAn scFv antibody-based multiplex assay has been developed capable of simultaneously measuring different proteins in a complex mixture, and to determine the potencies and compositions of allergen extracts.

Identification of Allergens of Dermatophagoides farinae on Canine Atopic Dermatitis in Korea

Dermatophagoides farinae plays important role in the pathogenesis of canine atopic dermatitis as environmental allergens. Also, many studies revealed that D. farinae was the main causative allergen for Korean dogs with atopic dermatitis. To identify major allergens of D. farinae in Korean atopic dogs allergic to D. farinae by immunoblot using commercial allergenic extracts, 26 dogs from two groups were enrolled in the study. Control group consists of 10 dogs with no clinical signs of disease and atopic group consists of 16 dogs diagnosed as atopic dermatitis. Sera from Korean dogs with atopic dermatitis showed six allergens of D. farinae extract by procedure of immunoblot. The molecular weights of identifying protein bands were 177, 109, 75, 44, 27, 15 kDa. The major allergens showing reactivity with greater than 50% of atopic dogs were detected at approximately 44, 109 and 177 kDa. Subsequent investigations will be carried out to verify the identity of the allergens detected in this study.

Characteristics of Sensitization to Inhalant and Food Allergens

<b>Background</b><b>:</b> Many studies worldwide have reported prevalence of atopic sensitization and its international variations. A positive skin prick reaction, however, does not always correlate with clinical symptoms. Positive skin tests (SPT) reveal rather atopy - specific IgE presence, than an atopic disease itself. When combined clinical manifestation and SPT are a powerful instrument for defining allergic diseases. Such data for Bulgaria are, however, scarce. The objective of the present study is to assess prevalence patterns and clinical relevance of sensitization to environmental and food allergens in a cross-sectional community sample in Bulgaria. <b>Methodology and results</b><b>:</b><i> </i>Patients, 225 men and women, age 4-81 years, were included. They completed a questionnaire to define asthma and rhinitis. Patients were skin prick tested to 18 commercial inhalant and food allergen extracts. The clinical relevance of each of the positively tested allergens was assessed. Among 225 patients 129 (57.3%) were sensitized to at least one allergen, using cut-off level ≥3 mm. In the inhalant allergens group the highest were rates of sensitization to 12 grasses – 26.6% and 4 cereals – 24.0%, followed by these to DFR – 18.2%, DPT – 17.8% and cockroach -16.4%. Among food allergens the highest rates of sensitizations were found to pork – 16.9%, walnut – 15.1%, apple – 13.3%, egg whole – 10.7%, celery – 9.8% and milk – 9.3%. The highest proportions of relevant tests in the inhalant allergens group were for DPT - 72.2%, DFR - 62.9%, 4 cereals - 69.2%, 12 grasses - 69%, Penicillium mix – 61.9, cockroach - 54.8%, Betulaceae - 52.4%. Among food allergens the highest proportions of relevant sensitization were found for walnut – 32.3%, peanut 29.4%, Milk - 22.2%, egg whole - 21.1%. <b>Conclusions</b><b>:</b><i> </i>We found rates and patterns of sensitization which were in line with data from other studies. The percentage of clinically relevant sensitizations differed significantly depending on the allergen.

Dual Function of Novel Pollen Coat (Surface) Proteins: IgE-binding Capacity and Proteolytic Activity Disrupting the Airway Epithelial Barrier

BackgroundThe pollen coat is the first structure of the pollen to encounter the mucosal immune system upon inhalation. Prior characterizations of pollen allergens have focused on water-soluble, cytoplasmic proteins, but have overlooked much of the extracellular pollen coat. Due to washing with organic solvents when prepared, these pollen coat proteins are typically absent from commercial standardized allergenic extracts (i.e., “de-fatted”), and, as a result, their involvement in allergy has not been explored.Methodology/Principal FindingsUsing a unique approach to search for pollen allergenic proteins residing in the pollen coat, we employed transmission electron microscopy (TEM) to assess the impact of organic solvents on the structural integrity of the pollen coat. TEM results indicated that de-fatting of Cynodon dactylon (Bermuda grass) pollen (BGP) by use of organic solvents altered the structural integrity of the pollen coat. The novel IgE-binding proteins of the BGP coat include a cysteine protease (CP) and endoxylanase (EXY). The full-length cDNA that encodes the novel IgE-reactive CP was cloned from floral RNA. The EXY and CP were purified to homogeneity and tested for IgE reactivity. The CP from the BGP coat increased the permeability of human airway epithelial cells, caused a clear concentration-dependent detachment of cells, and damaged their barrier integrity.Conclusions/SignificanceUsing an immunoproteomics approach, novel allergenic proteins of the BGP coat were identified. These proteins represent a class of novel dual-function proteins residing on the coat of the pollen grain that have IgE-binding capacity and proteolytic activity, which disrupts the integrity of the airway epithelial barrier. The identification of pollen coat allergens might explain the IgE-negative response to available skin-prick-testing proteins in patients who have positive symptoms. Further study of the role of these pollen coat proteins in allergic responses is warranted and could potentially lead to the development of improved diagnostic and therapeutic tools.

Evaluation of Banana Hypersensitivity Among a Group of Atopic Egyptian Children: Relation to Parental/Self Reports

PurposeTo evaluate the frequency of banana sensitization and allergy among a group of atopic Egyptian children in relation to parental/self reports.MethodsThis is a case-control study included 2 groups of allergic children with and without history of banana allergy, each included 40 patients. They were subjected to skin prick test (SPT) using commercial banana allergen extract and prick-prick test (PPT) using raw banana, in addition to measuring the serum banana-specific IgE. Oral banana challenge was performed in suspected cases.ResultsBanana allergy was diagnosed in 3 (7.5%) patients based on positive history of allergy on exposure to banana, positive SPT/PPT and elevated banana-specific IgE. The 3 patients had bronchial asthma with exacerbation upon banana exposure. The PPT results conform with those of SPT both in diagnosis of banana allergy and in the skin reactivity to banana. Serum banana-specific IgE was detectable in the whole studied sample with higher serum level among those without history of banana allergy (P=0.005). Oral banana challenge was negative for 20 patients with history of banana allergy and positive serum banana-specific IgE but negative SPT and PPT.ConclusionsSelf/parental reports of banana allergy is high while the actual banana allergy is uncommon. The PPT seems as reliable as SPT in diagnosis of banana allergy unlike specific IgE which reflects sensitization rather than allergy. Oral food challenge remains the most helpful tool for diagnosis of food allergy in suspected cases.

A Protein Allergen Microarray Detects Specific IgE to Pollen Surface, Cytoplasmic, and Commercial Allergen Extracts

BackgroundCurrent diagnostics for allergies, such as skin prick and radioallergosorbent tests, do not allow for inexpensive, high-throughput screening of patients. Additionally, extracts used in these methods are made from washed pollen that lacks pollen surface materials that may contain allergens.Methodology/Principal FindingsWe sought to develop a high-throughput assay to rapidly measure allergen-specific IgE in sera and to explore the relative allergenicity of different pollen fractions (i.e. surface, cytoplasmic, commercial extracts). To do this, we generated a protein microarray containing surface, cytoplasmic, and commercial extracts from 22 pollen species, commercial extracts from nine non-pollen allergens, and five recombinant allergenic proteins. Pollen surface and cytoplasmic fractions were prepared by extraction into organic solvents and aqueous buffers, respectively. Arrays were incubated with <25 uL of serum from 176 individuals and bound IgE was detected by indirect immunofluorescence, providing a high-throughput measurement of IgE. We demonstrated that the allergen microarray is a reproducible method to measure allergen-specific IgE in small amounts of sera. Using this tool, we demonstrated that specific IgE clusters according to the phylogeny of the allergen source. We also showed that the pollen surface, which has been largely overlooked in the past, contained potent allergens. Although, as a class, cytoplasmic fractions obtained by our pulverization/precipitation method were comparable to commercial extracts, many individual allergens showed significant differences.Conclusions/SignificanceThese results support the hypothesis that protein microarray technology is a useful tool for both research and in the clinic. It could provide a more efficient and less painful alternative to traditionally used skin prick tests, making it economically feasible to compare allergen sensitivity of different populations, monitor individual responses over time, and facilitate genetic studies on pollen allergy.

IgE cross-reactivity between Lolium multiflorum and commercial grass pollen allergen extracts in Brazilian patients with pollinosis

Lolium multiflorum (Lm) grass pollen is the major cause of pollinosis in Southern Brazil. The objectives of this study were to investigate immunodominant components of Lm pollen allergens and the cross-reactivity of IgE with commercial grass pollen allergen extracts. Thirty-eight serum samples from patients with seasonal allergic rhinitis (SAR), 35 serum samples from patients with perennial allergic rhinitis (PAR) and 30 serum samples from non-atopic subjects were analyzed. Allergen sensitization was evaluated using skin prick test and serum IgE levels against Lm pollen extract were determined by ELISA. Inhibition ELISA and immunoblot were used to evaluate the cross-reactivity of IgE between allergens from Lm and commercial grass pollen extracts, including L. perenne (Lp), grass mix I (GI) and II (GII) extracts. IgE antibodies against Lm were detected in 100% of SAR patients and 8.6% of PAR patients. Inhibition ELISA demonstrated IgE cross-reactivity between homologous (Lm) and heterologous (Lp or GII) grass pollen extracts, but not for the GI extract. Fifteen IgE-binding Lm components were detected and immunoblot bands of 26, 28-30, and 32-35 kDa showed >90% recognition. Lm, Lp and GII extracts significantly inhibited IgE binding to the most immunodominant Lm components, particularly the 55 kDa band. The 26 kDa and 90-114 kDa bands presented the lowest amount of heterologous inhibition. We demonstrated that Lm extract contains both Lm-specific and cross-reactive IgE-binding components and therefore it is suitable for measuring quantitative IgE levels for diagnostic and therapeutic purposes in patients with pollinosis sensitized to Lm grass pollen rather than other phylogenetically related grass pollen extracts.

Pyrethrins and Pyrethrosin Content in Commercial Allergen Extracts

Pyrethrins are the insecticidally active components of pyrethrum extract, derived from flowers of Chrysanthemum cinerariifolium and used in commercial and consumer insecticide products. Most dermal testing performed with pyrethrum extracts was done before current refined pyrethrum concentrate became available (before 1967). We analyzed presently commercially available pyrethrum allergen extracts to determine the concentration of pyrethrins and the putative sensitizer pyrethrosin. Six commercial pyrethrum allergen extracts were purchased from four major allergen suppliers and analyzed for pyrethrin I and pyrethrosin by using a capillary gas chromatograph equipped with a flame ionization detector. The commercial pyrethrum allergen extracts contained no detectable pyrethrins or pyrethrosin. In comparison, the pyrethrum standard provided by the McLaughlin Gormely King Company, a major refiner of pyrethrum, contained 20% pyrethrins and 0.49% pyrethrosin. No compounds observed in the chromatogram of the refined pyrethrum concentrate were present in the allergen extracts. Caution should be used when interpreting the results of tests performed with current pyrethrum allergen extracts because pyrethrins and pyrethrosin may not be present. Moreover, unknown components such as high-molecular-weight proteins or other impurities that may cause dermal reactions could be present in significant amounts.

Molecular composition and biological activity of commercial birch pollen allergen extracts

Commercial extracts used for diagnosis and treatment of allergy are currently prepared from natural allergen sources. The aim of this study was to analyse birch pollen allergen extracts produced for in vivo diagnosis of birch pollen allergy regarding their contents of individual birch pollen allergens (Bet v 1, Bet v 2 and Bet v 4). Protein contents were measured and the allergen composition was analysed by immunoblotting using antibody probes specific for Bet v 1, Bet v 2 and Bet v 4 in birch pollen extracts from five manufacturers of allergen extracts. The contents of the major birch pollen allergen, Bet v 1, were quantified with a specific two-site binding enzyme-linked immunosorbent assay with nanogram sensitivity for Bet v 1. The biological activities of the allergen extracts were evaluated by skin prick testing in birch pollen allergic patients and compared with their sensitization profiles. A more than 10-fold variation regarding total protein contents (23.1-314 microg mL(-1)) and also regarding the amounts of the major birch pollen allergen, Bet v 1 (1.62-19.6 microg mL(-1)) was found. The highly cross-reactive Bet v 4 allergen was absent in three of the five tested extracts. Furthermore, varying skin test results were obtained in birch pollen allergic patients with the allergen extracts. Commercial birch pollen extracts exhibit a considerable variability regarding allergen contents and hence deliver varying in vivo test results. These problems might be overcome with recombinant allergen-based preparations.

Exposure to allergens of different cattle breeds and their relevance in occupational allergy

IntroductionCattle are an important source of allergens in the working area of farmers. Asthma caused by cow allergens is a significant occupational problem. Yet in allergological testing, the results of in vivo and in vitro diagnostic tests are often inconsistent even in cases with clearly cattle-related symptoms.Objectives and methodsThe aim of this study was to investigate four different commercial cow allergen extracts and to compare them with self prepared extracts of different cattle breeds by means of SDS-PAGE and immunoblotting using the sera of 42 German farmers with asthma and rhino-conjunctivitis caused by cattle contact.ResultsThe commercial extracts investigated in this study showed only minor differences in protein pattern. Using sera in immunoblotting experiments distinct bands were found for all symptomatic farmers, even in 13 farmers with a negative result in commercially available serological allergy tests. Bands with molecular weights in the range between about 11 and 67 kDa were observed; reactivity with the major allergen Bos d 2 at about 20 kDa was detected in all farmers, although it was not the strongest band in all cases.ConclusionsWe demonstrate for the first time the allergenic relevance of additional proteins with molecular weights of 14, 30, 55 and approx. 67–97 kDa in more than 50% of farmers with cattle related symptoms. One of our most striking results was that 32% of the investigated farmers with cattle related symptoms showed negative results with commercial serological tests but distinct reactions with cow allergen in immunoblotting experiments. The Bos d 2 content in hair showed differences between certain breeds whereas German Brown and Simmental had particularly higher quantities of Bos d 2 in their hair than breeds such as Holstein-Friesian. These results strongly support the following recommendation: test results with commercial extracts that are contradictory to the clinical symptoms should be supplemented by skin tests using extracts of the hair of the farmers’ own cattle.

Heterogeneity of commercial timothy grass pollen extracts

The diagnosis and specific immunotherapy of allergy is currently performed with allergen extracts prepared from natural allergen sources. To analyse commercial timothy grass pollen allergen extracts used for in vivo diagnosis regarding their qualitative and quantitative allergen composition and in vivo biological activity. Antibodies specific for eight timothy grass pollen allergens (Phl p 1, Phl p 2, Phl p 4, Phl p 5, Phl p 6, Phl p 7, Phl p 12, Phl p 13) were used to detect these allergens in timothy grass pollen extracts from four manufacturers by immunoblotting. ELISA assays were developed and used to quantify the three major allergens (Phl p 1, Phl p 2, Phl p 5) in the extracts. The magnitude of skin responses to the four extracts was studied by skin prick testing in 10 grass pollen-allergic patients. The allergen extracts showed broad variations in protein compositions and amounts (24.1-197.7 microg/mL extract). Several allergens could not be detected in certain extracts or appeared degraded. A considerable variability regarding the contents of major allergens was found (Phl p 1: 32-384 ng/mL; Phl p 2: 1128-6530 ng/mL, Phl p 5: 40-793 ng/mL). Heterogeneous skin test results were obtained with the extracts in grass pollen-allergic patients. Timothy grass pollen extracts from different manufacturers exhibit a considerable heterogeneity regarding the presence of individual allergens and hence yield varying in vivo test results. Problems related to the use of natural grass pollen allergen extracts may be circumvented by using defined recombinant grass pollen allergens.

Environmental Exposure to Allergens of Different Dog Breeds and Relevance in Allergological Diagnostics

In our environment, dogs are a relevant source of allergens, but diagnosing dog-related allergies may present difficulties, as in diagnostic tests with commercial dog allergens, some patients show only slight positive or negative results, even though they suffer from dog-related symptoms. Occasionally, allergy tests with extracts of dog hair belonging to patients' dogs or from dogs of the same breed were found to yield more reliable results, possibly due to breed-specific allergen components. The purpose of this study was to determine breed-specific differences or possibly hypo- or hyperallergenic dog breeds. The dog allergen content and protein patterns of different commercial and self-prepared dog allergen extracts were compared. Protein extracts were separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and stained with silver. The major allergen Can f 1 was quantified using the commercially available enzyme-linked immunosorbent assay (ELISA) technique. The majority of the bands in the self-prepared extracts of different breeds had a molecular mass lower than 30 kD. Notably, the self-prepared extracts of hair of common breeds showed distinct protein bands with a molecular mass lower than 14 kD, which the commercial extracts did not. With regard to Can f 1 content, a marked variability occurred. Factors related to individual dogs seem to influence the allergenicity more than breed or gender. This is the first report to describe allergens with low molecular mass that are absent in extracts of commercial test kits. Consequently, skin tests with self-prepared dog allergen extracts need to be performed in case of inconsistent test results with commercial extracts.

A comparison of intradermal testing and detection of allergen-specific immunoglobulin E in serum by enzyme-linked immunosorbent assay in horses affected with skin hypersensitivity

Skin hypersensitivities (allergies) in horses are often diagnosed using clinical signs only. Intradermal testing or serological assays are diagnostic options to confirm the allergic nature of the disease and to identify the allergen(s). Our objective was to develop an allergen-specific enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody specific for horse IgE and to examine its potential for allergen detection in serum in comparison to intradermal testing. Intradermal testing with 61 allergen extracts was performed on 10 horses affected with skin hypersensitivity. Their sera were analyzed by ELISA for IgE antibodies to the same allergens. The kappa test of concordance was used for comparison of the results of both tests. Out of 61 allergen extracts, only two (Timothy and Quack) had kappa values greater than 0.60, suggesting a substantial agreement between skin testing and IgE ELISA. The statistical comparison of the remaining 59 allergens showed little or no concordance between the tests beyond chance. To identify parameters that may influence the sensitivity of the ELISA, the assay was modified to detect allergen-specific IgGb and IgG(T) in serum, and the protein content in all allergen extracts was determined by SDS-PAGE. The commercial allergen extracts revealed a high variation in detectable protein. High concentrations of allergen-specific IgG in horse serum were found to compete with IgE for binding to the plates. In conclusion, an ELISA using whole serum and crude allergen preparations provides limited diagnostic information in horses. The reliable diagnosis of allergens in equine skin hypersensitivity is essential to improve allergen-specific treatments, such as hyposensitization, or the development of allergy vaccines.

Harmonia axyridis ladybug hypersensitivity in clinical allergy practice

The imported Harmonia axyridis ladybug infests homes in northern West Virginia from fall through spring, causing allergic disease. Retrospective single-practice chart reviews were performed: (1) all skin prick tests (1400 included ladybug) in a community allergy practice over 4 years and (2) clinical analysis of 400 randomly chosen patients. The usual adult aeroallergen skin test panel included ladybug and 57 other allergens. Statistics used were contingency table analyses and the kappa-statistic for concordance. Home infestation with ladybugs was most common in rural areas but did not predict ladybug sensitization (kappa = -0.02). Ladybug sensitization and allergy occurred at all ages. Ladybug sensitization occurred with 21% frequency compared with cat at 24% frequency, cockroach at 27% frequency, and dust mites at 40% frequency. Only ladybug showed a significant (p < 0.0001) skin test sensitization decreasing from rural (30%), mixed (21%), to urban (16%) home demographics. Isolated single-positive skin tests constituted 10% of dust mites, 6% of cockroach, 6% of ladybug, and 4% of cat-positive skin tests. Skin test concordance was strongest between the pairs: ladybug-cockroach (kappa = 0.36), cockroach-dust mite (kappa = 0.29), and dust mite-cat (kappa = 0.25). Ladybug is a major allergen in endemic areas, causing rhinoconjunctivitis (8% prevalence), asthma (2% prevalence), and urticaria (1% prevalence). Ladybug skin test sensitization is more common in rural areas and is comparable in frequency and age distribution with cat and cockroach. Cockroach and ladybug have a high degree of skin test concordance. A quality commercial ladybug allergen extract and increased ladybug allergen research are needed.

Risk Factors for Sensitization to Anisakis Simplex: A Multivariate Statistical Evaluation

Anisakis simplex is a nematode belonging to the Anisakidae family. The ingestion of third stage larvae in uncooked or undercooked seafood may cause human diseases known as anisakiasis and anisakidosis. A total of 400 (159 atopic and 241 non-atopic) subjects living in an area of southern Italy (Bari district) were consecutively evaluated to identify the association of some factors (sex, age, atopy, consumption of uncooked seafood and sensitization to dust mites) with the risk of Anisakis simplex sensitization. Patients were investigated on history of atopy and allergic diseases and were skin prick tested with commercial allergen extracts of Anisakis simplex, Acarus siro, Lepidoglyphus destructor, Tyrophagus putrescentiae, Glycyphagus domesticus, Euroglyphus maynei, Dermatophagoides pteronyssinus and Dermatophagoides farinae. Our results suggest that atopic subjects have a lower risk of Anisakis allergy than non-atopic subjects and show the association of Anisakis simplex sensitization with the consumption of uncooked seafood (anchovies and squid), increasing age and sensitization to Glycyphagus domesticus.

Harmonia axyridis Ladybug Hypersensitivity in Clinical Allergy Practice

The imported Harmonia axyridis ladybug infests homes in northern West Virginia from fall through spring, causing allergic disease. Retrospective single-practice chart reviews were performed: (1) all skin prick tests (1400 included ladybug) in a community allergy practice over 4 years and (2) clinical analysis of 400 randomly chosen patients. The usual adult aeroallergen skin test panel included ladybug and 57 other allergens. Statistics used were contingency table analyses and the -statistic for concordance. Home infestation with ladybugs was most common in rural areas but did not predict ladybug sensitization ( 0.02). Ladybug sensitization and allergy occurred at all ages. Ladybug sensitization occurred with 21% frequency compared with cat at 24% frequency, cockroach at 27% frequency, and dust mites at 40% frequency. Only ladybug showed a significant (p 0.0001) skin test sensitization decreasing from rural (30%), mixed (21%), to urban (16%) home demographics. Isolated single-positive skin tests constituted 10% of dust mites, 6% of cockroach, 6% of ladybug, and 4% of cat-positive skin tests. Skin test concordance was strongest between the pairs: ladybug–cockroach ( 0.36), cockroach–dust mite ( 0.29), and dust mite–cat ( 0.25). Ladybug is a major allergen in endemic areas, causing rhinoconjunctivitis (8% prevalence), asthma (2% prevalence), and urticaria (1% prevalence). Ladybug skin test sensitization is more common in rural areas and is comparable in frequency and age distribution with cat and cockroach. Cockroach and ladybug have a high degree of skin test concordance. A quality commercial ladybug allergen extract and increased ladybug allergen research are needed. (Allergy Asthma Proc 28:50–57, 2007; doi: 10.2500/aap.2007.28.2956)