Analysis of IgE binding proteins of mesquite (Prosopis juliflora) pollen and cross-reactivity with predominant tree pollens

Pollen from the mesquite tree, Prosopis juliflora, is an important source of respiratory allergy in tropical countries. Our aim was to partially characterize the IgE binding proteins of P. juliflora pollen extract and study cross-reactivity with prevalent tree pollen allergens. Intradermal tests with P. juliflora and five other tree pollen extracts were performed on respiratory allergy patients from Bikaner (arid) and Delhi (semi arid). Prosopis extract elicited positive skin reactions in 71/220 of the patients. Sera were collected from 38 of these 71 patients and all demonstrated elevated specific IgE to P. juliflora. Immunoblotting with pooled patients' sera demonstrated 16 IgE binding components, with components of 24, 26, 29, 31, 35, 52, 58, 66 and 95 kDa recognized by more than 80% of individual patients' sera. P. juliflora extract is allergenically potent requiring 73 ng of self-protein for 50% inhibition of IgE binding in ELISA inhibition. Cross-inhibition assays showed close relationship among P. juliflora, Ailanthus excelsa, Cassia siamea and Salvadora persica. IgE binding components of 14, 41, 52 and 66 kDa were shared allergens whereas 26 and 29 kDa were specific to P. juliflora. The findings suggest that purification of cross-reactive allergens will be helpful for diagnosis and immunotherapy of tree pollen allergic patients.     

Purification and partial characterization ofa 67-kD cross-react ive allergen from Imperata cylindrica pollen extract

BACKGROUND: Grass pollens are known to induce type I allergic reactions in a large number of genetically predisposed individuals. Earlier studies have recognized Imperata cylindrica (Ic) pollen as an important source of aeroallergen which contained 7 IgE binding proteins in the MW range of 85-16 kD. OBJECTIVES: To isolate, purify and characterize a cross-reactive allergenic protein from Ic pollen extract for diagnosis and therapy of grass pollen allergy. METHODOLOGY: Ic pollen extract was fractionated using DEAE Sephadex A-50, Sephadex G-200 and Mono Q column. Allergenic activity of the fractions was checked by ELISA, skin tests, ELISA inhibition and immunoblot using sera of Ic-sensitive patients. A 67-kD protein was purified to homogeneity from Ic-VIII. The allergenic determinants of this protein were identified by SDS-PAGE and immunoblot after CNBr treatment. RESULTS: Among Ic fractions, Ic-VIII was highly potent by ELISA, skin tests and showed cross-reactivity with 4 other tropical grasses by immunoblot and ELISA inhibition. The subfraction Ic-VIIIe1 of Ic-VIII showed a band at 67 kD on SDS-PAGE. On CNBr treatment, it gave 7 peptides, 3 of which were found to be allergenic. CONCLUSION: A 67-kD protein (Ic-VIIIe1) was isolated, purified to homogeneity and partially characterized. It showed cross-reactivity with tropical grasses tested and contained at least three allergenic determinants.     

Assessment of cross-reactivity between Japanese cedar (Cryptomeria japonica) pollen and tomato fruit extracts by RAST inhibition and immunoblot inhibition

BACKGROUND: An association between pollinosis and sensitivity to fruits and vegetables has been reported. Although Japanese cedar (Cryptomeria japonica) pollinosis is one of the most widespread diseases in Japan, there have been no reports demonstrating cross-reactivity between Japanese cedar pollen and other plant food. OBJECTIVE: The aim of this study was to demonstrate cross-reactivity between Japanese cedar pollen and tomato fruit (Lycopersicon esculentum) using RAST inhibition and immunoblot inhibition. METHODS: The RAST and immunoblot inhibition were performed using sera from patients with oral allergy syndrome (OAS) after ingesting fresh tomatoes. We identified some proteins that took part in cross-reactive IgE by the determination of N-terminal amino acid sequences and a homology search through the SWISS-PROT database. RESULTS: In the RAST inhibition, the bindings of IgE from the sera from four out of five (4/5) subjects to Japanese cedar pollen discs were inhibited by more than 50% by preincubation of the serum with tomato fruit extracts. Likewise, the IgE bindings to tomato fruit discs were inhibited more than 50% by Japanese cedar pollen extracts in 3/5 sera. In immunoblot inhibition, IgE binding activities of some protein bands on both membranes were decreased by heterologous inhibitors. However, the combinations of these protein bands involved in cross-reactivity were different between patients. CONCLUSION: We have demonstrated cross-reactivity between Japanese cedar pollen and tomato fruit using RAST inhibition and immunoblot inhibition.     

IgE immune response to Ginkgo biloba pollen.

BACKGROUND: The ginkgo (Ginkgo biloba L.) continues to be planted as a shade tree in preference to other species in Seoul, Korea. The proportion of ginkgo to total shade trees was 43.2% in 1998, but the allergenic characteristics of ginkgo pollen has not been elucidated. OBJECTIVES: This study was undertaken to obtain information regarding the skin reactivity rate to ginkgo pollen in a population of Korean subjects with respiratory allergy. Possible ginkgo pollen allergens and the cross-reactivity of ginkgo pollen with other prevalent pollens were also examined. METHODS: Four hundred and forty-seven patients with asthma and/or allergic rhinitis were skin prick tested with extract of ginkgo pollen (1:20 wt/vol). Of these patients, positive skin responders (A/H ratio > or =2+) were selected for ELISA and immunoblot experiments. RESULTS: A total of 21 patients (4.7%) showed skin reactivity (A/H ratio > or =2+) to ginkgo pollen in the skin prick test. They were also cosensitized to many other tree, grass, and weed pollens. Sixteen (76%) of the 21 positive skin responders showed specific IgE responses to ginkgo pollen in ELISA. In inhibitory ELISA, IgE binding to ginkgo pollen was inhibited by more than 80% by oak, ryegrass, mugwort, and ragweed; and 34% by hop Japanese; and 10% by rBet v 2 at 10 microg/mL. In immunoblot, 10 out of 21 sera (48%) reacted to the 15-kD protein of ginkgo pollen, 9 (43%) to 33-35 kD, and 8 (38%) to 36-38 kD. In inhibitory immunoblot, IgE binding to ginkgo pollen proteins was almost completely inhibited by oak, ryegrass, mugwort and ragweed, but only partially by hop Japanese and rBet v 2. CONCLUSION: The skin reactivity rate to ginkgo pollen is approximately 4.7% in a population of Korean subjects with respiratory allergy. Since ginkgo pollen has a high cross-reactivity with other prevalent pollens, it could cause clinical symptoms during its pollen season by cross-reacting with the IgE produced in response to other pollens in patients sensitized to multiple pollens.     

Sensitization to Lolium multiflorum grass pollen in pollinosis patients: evaluation of allergenic fractions recognized by specific IgE antibodies

BACKGROUND: Lolium multiflorum (Lm) pollen allergens are the major causative agents for rhinoconjunctivitis in Southern Brazil. There have been no studies about the sensitization and allergenic cross-reactivity between Lm and other grass pollens. We evaluated the sensitization of Brazilian pollinosis patients to Lm pollen allergens through skin prick test (SPT) and immunoassays (ELISA and immunoblot). METHODS: Serum samples from 60 patients with pollinosis and positive SPT to grass pollen extracts (Lm+ group), 30 patients with negative SPT to grass pollen, but positive SPT to mite extracts (Lm- group), and 30 nonatopic subjects (NA group) were tested by SPT, ELISA, and immunoblot using Lm extract. Inhibition immunoassays with Lolium perenne (Lp), mixed grass (Gmix) and Lm extracts were also performed. RESULTS: A high concordance was found between the Gmix and Lm extracts in SPT. Positivity rates in SPT were also highly concordant with IgE-ELISA results. The assay was able to detect Lm-specific IgE in >95% of Lm+ patients. A significant self- and cross-inhibition was observed in IgE-ELISA, reflecting a high cross-reactivity between the grass pollen allergens. Immunoblot revealed 13 IgE-binding Lm fractions, from which the bands 28-30 kDa and 31-34 kDa were recognized by >90% of Lm+ patients. CONCLUSION: Lm-specific IgE antibodies are highly cross-reactive with pollen proteins from other grass species. The results indicate that Lm extracts could be used in both SPT and ELISA for a more specific evaluation of IgE responses to Lm grass pollen in Brazilian pollinosis patients.     

Antigenic and allergenic cross-reactivity of Epicoccum nigrum with other fungi.

BACKGROUND: Previous studies have identified Epicoccum nigrum (EN) as an important aeroallergen. Shared allergenicity among some fungi responsible for type I allergic disorders has been reported. OBJECTIVE: To study the cross-reactivity among different fungi and to identify immunoglobulin (Ig)G and IgE binding components shared between EN and 10 other fungi known to cause respiratory allergy. METHODS: Cross-reactivity studies were carried out by enzyme-linked immunoadsorbent assay (ELISA) and immunoblot inhibition using both rabbit antiserum raised against EN and pooled sera from patients' EN-positive skin test. RESULTS: A large number (82%) of EN-sensitive patients showed positive skin reactivity to other fungal extracts. ELISA inhibition revealed >50% inhibition in binding of EN-specific rabbit antibodies with Alternaria alternata, Curvularia lunata, Cladosporium herbarum, and Penicillium citrinum extract, whereas the other extracts showed only 20 to 40% inhibition. Rabbit antisera to A. alternata, C. herbarum, and C. lunata reacted with five to seven bands in EN, demonstrating the presence of shared antigens among these fungi. EN requires an amount of 100 ng for 50% IgE ELISA inhibition, whereas 175 ng of A. alternata, 160 ng of C. lunata, and 268 ng of C. herbarum and P. citrinum were required for the same. IgE immunoblot and immunoblot inhibition further revealed that 43-, 26-, and 17-kD allergenic bands were shared by EN and A. alternata, whereas the 80- and 37-kD bands were common to both EN and C. lunata. EN and C. herbarum shared 63- and 36-kD allergenic bands, whereas EN and P. citrinum shared the 34-kD band. CONCLUSION: EN showed maximum cross-reactivity with A. alternata followed by C. lunata, C. herbarum, and P. citrinum. This information will be useful in treating EN-allergic patients.     

The major allergen of olive pollen Ole e 1 is a diagnostic marker for sensitization to Oleaceae

BACKGROUND: Trees of the family Oleaceae are important allergen sources, with a strongly varying geographic distribution. For example, olive pollen is an important allergen source in Mediterranean countries, whereas ash pollen dominates in Northern and Central Europe and North America. The aim of this study was to compare the profiles of olive and ash pollen allergens and to study the degree of cross-reactivity using populations of allergic patients selectively exposed to olive or ash pollen. METHODS: Olive and ash pollen extracts were analyzed by IgE immunoblotting using sera from Spanish patients highly exposed to olive pollen and Austrian patients without olive but ash pollen exposure. IgE cross-reactivity was studied by qualitative immunoblot inhibition assays and semiquantitative ELISA inhibitions using olive, ash, birch, mugwort, timothy grass pollen extracts and the major olive pollen allergen, Ole e 1. RESULTS: Spanish and Austrian patients exhibited an almost identical IgE-binding profile to olive and ash pollen allergens, with major reactivity directed against Ole e 1, and its homologous ash counterpart, Fra e 1. IgE inhibition experiments demonstrated extensive cross-reactivity between olive and ash pollen allergens. However, whereas cross-reactions between profilins and calcium-binding allergens also occurred between unrelated plant species, cross-reactivity to Ole e 1 was confined to plants belonging to the Oleaceae. CONCLUSIONS: Ole e 1 is a marker allergen for the diagnosis of olive and ash pollen allergy.     

Patients with anaphylaxis to pea can have peanut allergy caused by cross-reactive IgE to vicilin (Ara h 1)

BACKGROUND: Serologic cross-reactivity among legumes has been described; however, it is rarely clinically significant. In this study 3 patients with a history of anaphylaxis to pea are described who subsequently had symptoms after ingestion of peanut. OBJECTIVE: We investigated whether the peanut-related symptoms were due to cross-reactivity between pea and peanut proteins. METHODS: Peanut-related symptoms were documented according to case history or double-blind, placebo-controlled food challenge results. Skin prick tests were performed, and specific IgE levels were determined for pea and peanut with the CAP system FEIA. IgE-binding proteins in pea and peanut were identified by using immunoblot analysis. Cross-reactivity was studied by means of immunoblot and ELISA inhibition studies with whole extracts and purified allergens. RESULTS: Peanut-related symptoms consisted of oral symptoms in all patients, with additional urticaria and dyspnea or angioedema in 2 patients. All patients had a positive skin prick test response and an increased IgE level to pea and peanut. Immunoblotting revealed strong IgE binding to mainly vicilin in pea extract and exclusively to Ara h 1 in crude peanut extract. Immunoblot and ELISA inhibition studies with crude extracts, as well as purified proteins, showed that IgE binding to peanut could be inhibited by pea but not or only partially the other way around. CONCLUSION: Clinically relevant cross-reactivity between pea and peanut does occur. Vicilin homologues in pea and peanut (Ara h 1) are the molecular basis for this cross-reactivity.     

Cladosporium herbarum and Pityrosporum ovale allergen extracts share cross-reacting glycoproteins

BACKGROUND: Patients sensitized to airborne fungi such as Alternaria alternata and Cladosporium herbarum often also show positive skin prick test results and specific serum IgE antibodies to a yeast, Pityrosporum ovale. We examined whether part of the IgE binding to these fungi is explained by cross-reacting mould and yeast allergens. METHODS: Serum samples from 36 patients with positive skin prick test to A. alternata or C. herbarum were analyzed for IgE antibodies to fungal extracts by ELISA and immunoblot analysis. Cross-reactivity between mould and yeast extracts was studied by ELISA and immunoblot inhibition assays. In further analysis, the mannan-containing glycoproteins were removed from the yeast extract by concanavalin A-Sepharose chromatography, and the IgE binding properties of the extracts were compared. RESULTS: Serum IgE reactivity to P. ovale was found in 40% of the mould-sensitized patients. The IgE antibody binding to A. alternata and C. herbarum moulds was partially inhibited by the yeast P. ovale in ELISA and immunoblot inhibition assays. When the glycoproteins were removed from the extract, cross-reactivity was markedly reduced. CONCLUSION: Part of the IgE binding to mould and yeast allergen extracts is due to cross-reacting glycoproteins. False-positive IgE and skin prick test results should be taken into account in the diagnosis of mould allergy.     

A Case of Occupational Rhinitis Induced by Maize Pollen Exposure in a Farmer: Detection of IgE-Binding Components

Corn is a major staple food, along with rice and wheat, in many parts of the world. There are several reports of hypersensitivity to maize pollen. However, cases of occupational allergic rhinitis induced by inhalation of maize pollen are very rare. We herein report the case of a 67-year-old male with occupational rhinitis caused by occupational exposure to maize pollen in a cornfield. He showed positive responses to maize pollen, as well as grass pollens, in skin prick tests. A high level of serum immunoglobulin E (IgE) specific to maize pollen extracts was detected by an enzyme-linked immunosorbent assay (ELISA). Laboratory tests showed a high serum level of total IgE (724 kU/L) and a high level of IgE specific to maize pollen (8.32 kU/L) using the Immuno-CAP system. Occupational rhinitis was confirmed by a nasal provocation test with maize pollen extracts. IgE ELISA inhibition tests showed antibody cross-reactivity between maize pollen and grass pollen extracts. IgE immunoblotting using maize pollen extracts demonstrated a 27 kDa IgE-binding component. These findings suggest that maize pollen can induce IgE-mediated occupational rhinitis in exposed workers.     

Occupational Allergy to Flowers: Immunoblot Analysis of Allergens in Freesia,Gerbera and Chrysanthemum Pollen

High exposure to pollen from ornamental flowers can induce an IgE‐mediated occupational allergy in florists and horticulture workers. We investigated IgE‐binding antigens in chrysanthemum, freesia and gerbera pollen by immunoblot analysis and analysed the cross‐reactivity of these pollen with birch, grass and mugwort pollen. In immunoblots with chrysanthemum pollen, major IgE‐binding structures were seen with a molecular weight (MW) of approximately 25, 45 and 65 kD. In the immunoblots with freesia pollen, IgE from freesia pollen was directed against two proteins with an MW of approximately 15 kD. Most sera showed IgE binding to an approximately 15 kD band in gerbera pollen; with some sera additional bands were seen in the range of 30–50 kD. IgE binding to chrysanthemum pollen was inhibited by mugwort pollen only, whereas IgE binding to freesia pollen was suppressed by birch, grass and mugwort pollen. The inhibitory activity of birch and grass pollen extract on IgE binding to gerbera pollen extract was serum dependent and ranged from no inhibition to complete inhibition. Occupational exposure to many different flowers induced IgE against all three types of pollen. Exposure in greenhouses to gerbera flowers elicited mainly IgE against gerbera pollen. Mugwort pollen extract inhibited IgE binding to pollen from all three flowers.     

Primary sensitization to sweet bell pepper pollen in greenhouse workers with occupational allergy

BACKGROUND: In a previous investigation, a high prevalence of allergy to sweet bell pepper pollen was found among exposed horticulture workers. Allergy to plant-derived food is often the consequence of primary sensitization to common pollen allergens. OBJECTIVE: We therefore investigated the cross-reactivity between sweet bell pepper pollen and pollen from grass, birch or mugwort. METHOD: We selected 10 sera from greenhouse workers who had, besides specific IgE against sweet bell pepper pollen, also IgE to grass, birch or mugwort pollen. Cross-reactivity was tested by the inhibition of IgE binding to solid-phase coupled sweet bell pepper pollen extract. The 10 sera were also analysed for IgE binding to sweet bell pepper pollen by immunoblotting. RESULTS: With these sera, no or small inhibition of IgE binding to sweet bell pepper pollen extract was observed with grass, birch and mugwort pollen. With immunoblotting, major IgE-binding structures were seen at 14, 29 and 69 kDa in sweet bell pepper pollen extract. CONCLUSION: The results of our study demonstrate that sweet bell pepper pollen contains allergens that have no or limited cross-reactivity with common pollen allergens. With sera from the 10 patients tested, sensitization to sweet bell pepper pollen was not the consequence of primary sensitization to common pollen allergens.     

Allergenic cross-reactivity of olive pollen

BACKGROUND: Sera of patients allergic to olive (Olea europaea) pollen were used to analyze the IgE cross-reactivity between olive-pollen extract and other pollens obtained from phylogenetically unrelated species. METHODS: We used IgE immunostaining of pollen extracts blotted to nitrocellulose membranes after SDS-PAGE and inhibition analysis of this binding. RESULTS: A high inhibition of the IgE binding on olive-pollen extract was exhibited by birch, mugwort, pine, and cypress pollens, suggesting that these extracts contain proteins which share common epitopes and thus can be recognized by olive-allergic sera. IgE binding to Gramineae pollen extracts was not inhibited by olive-pollen extract, indicating a primary sensitization of the patients to these species. From the inhibition assays, the presence of an allergen of 45 kDa in the olive pollen, which has no homologous counterparts in other allergenic species, has been inferred. CONCLUSIONS: Olive pollen contains allergens which cross-react with pollens from unrelated species, a fact that could simplify the diagnosis and treatment of pollinosis.     

Ara h 8, a Bet v 1-homologous allergen from peanut, is a major allergen in patients with combined birch pollen and peanut allergy

BACKGROUND: We recently described patients with soybean allergy mainly mediated by cross-reactivity to birch pollen allergens. A majority of those patients were reported to have peanut allergy. OBJECTIVE: We sought to study the occurrence of peanut allergy in patients allergic to birch pollen and characterized the Bet v 1-homologous peanut allergen Ara h 8. METHODS: Recombinant Ara h 8 was cloned with degenerated primers and expressed in Escherichia coli. Nine Swiss and 11 Dutch patients with peanut and birch pollen allergy and a positive double-blind, placebo-controlled food challenge result to peanut were investigated for IgE reactivity to birch pollen and purified peanut allergens and cross-reactivity between birch and peanut. Ara h 8 stability against digestion and roasting was assessed by means of RAST inhibition. The IgE cross-linking potency of Ara h 8 was tested on the basis of basophil histamine release. RESULTS: During double-blind, placebo-controlled food challenge, all patients experienced symptoms in the oral cavity, progressing to more severe symptoms in 40% of patients. CAP-FEIA detected recombinant (r) Ara h 8-specific IgE in 85%. IgE binding to Ara h 8 was inhibited by Bet v 1 in peanut extract immunoblotting and in RAST inhibition. In EAST inhibition recombinant rAra h 8 inhibited IgE binding to peanut in 4 of 7 tested patient sera. Antipeanut response was dominated by Ara h 8 in 12 of 17 tested patients. Furthermore, our results demonstrate a low stability of Ara h 8 to roasting and no stability to gastric digestion. Basophil histamine release with rAra h 8 was more than 20% in 5 of 7 tested sera. CONCLUSIONS: Peanut allergy might be mediated in a subgroup of our patients by means of cross-reaction of Bet v 1 with the homologous peanut allergen Ara h 8.     

Identification of a 7S globulin as a novel coconut allergen.

BACKGROUND: Coconut (Cocos nucifera) is a monocotyledonous plant of the Arecaceae family. Allergy to coconut is infrequent, with only 5 cases reported so far in the medical literature. OBJECTIVE: To identify coconut allergens in 2 patients allergic to this food. METHODS: We describe 2 patients allergic to coconut: an adult pollen-allergic patient monosensitized to coconut who presented with severe oropharyngeal symptoms and a child with a previous allergy to walnut, not allergic to pollen, who developed anaphylaxis on coconut ingestion. Both patients had positive skin prick test results and serum specific IgE (CAP) to coconut. IgE sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting was performed to identify the allergens involved, and a strong IgE binding band detected in both patients was further analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS). Stability to pepsin digestion of the coconut extract and its cross-reactivity with tree nuts were studied. RESULTS: An immunoblot showed an almost identical profile of IgE binding proteins in the coconut extract in both patients who reacted strongly to a band of approximately 29 kDa. The peptide analysis by MALDI-TOF MS of this band obtained the sequence GHGKREDPEKR. The protein with the highest correlation with this peptide was found to be a 7S globulin from Elaeis guineensis, another oil palm species also belonging to the Arecaceae family. The 29-kDa band was digested by pepsin in less than 1 minute. Cross-reactivity among coconut, walnut, and hazelnut was demonstrated by CAP inhibition in patient 2. CONCLUSION: We have identified a 7S storage protein as a novel coconut allergen.     

Evidence of cross-reactivity between olive, ash, privet, and Russian olive tree pollen allergens.

In a clinical investigation, 103 Michigan residents with symptoms suggestive of allergic rhinitis or asthma were skin tested with olive (Olea europaea) pollen extract. Nineteen had positive reactions. Since the olive tree is not native to nor grown in Michigan, this study was undertaken to determine whether the skin test reactivity was the result of cross-reactivity among tree pollen allergens. ELISAs were developed to measure olive, ash (Fraxinus americana), privet (Ligustrum vulgare), and Russian olive (Elaeagnus angustifolia) specific IgE antibodies. Inhibition studies were performed to determine whether pollen extracts from each of these tree species could inhibit IgE antibody binding to olive extracts. Eleven of the 19 skin test-positive patients were olive-ELISA positive, eight either were ELISA-positive to ash, seven to privet and ten to Russian olive. There were significant correlations between the ELISA results to olive and each of the other three pollens. The inhibition studies demonstrated that all three of the tree pollens were capable of inhibiting the binding of IgE to olive extract in a dose-response fashion. IgE-immunoblot studies demonstrated several proteins common to olive, ash, and privet. Twelve of the olive skin test-positive patients were contacted and 75% were exposed to one or more of the studied trees in their yards. Five patients had traveled to areas where olive trees are grown. We conclude that there is a high degree of cross-reactivity among allergens from native Michigan trees and from olive trees. This cross-reactivity is the most likely reason for skin test reactivity to olive pollen extract in Michigan.     

Immunoblot analysis of IgE-binding antigens in paprika and tomato pollen

BACKGROUND: The high incidence of occupational allergy in horticulture has only recently been recognized. We determined IgE against pollen and fruit from paprika and tomato plants in sera from 3 greenhouse workers and in 3 sera from food-allergic patients. METHODS: Proteins in extracts of paprika and tomato pollen were incubated with patients' sera after covalent coupling of these proteins to agarose beads, or in immunoblots. RESULTS: IgE against paprika pollen, but no IgE against tomato pollen, was found in serum from 2 greenhouse workers who worked with paprika plants only. IgE binding of these 2 sera to agarose-bound paprika pollen extract could be inhibited by paprika pollen but not by tomato pollen extract. A greenhouse worker, who cultivated tomato plants, had IgE against both tomato and paprika pollen. IgE binding of this serum to agarose-bound paprika pollen extract could be inhibited by both paprika pollen and tomato pollen extract. Three food-allergic patients also had IgE against tomato and paprika pollen. IgE from 2 food-allergic patients recognized IgE-binding structures in paprika or tomato pollen that were also present in fruit from the corresponding plant. In contrast, no substantial cross-reactivity was observed between paprika pollen and fruit towards IgE from 3 greenhouse workers. In 4 of 5 sera that were positive in the paprika pollen immunoblot major IgE binding to allergens of about 30 and 64 kD occurred. CONCLUSION: The presence of IgE against paprika or tomato pollen is not restricted to workers in horticulture; IgE against these pollen can also be present in food-allergic patients who have serum IgE against paprika and/or tomato fruit.     

Analysis of IgE binding proteins of Celtis tala pollen

Celtis tala is a tree, widely distributed in Argentina. Its significance as a source of allergic sensitisation is still unclear. Our aim was to partially characterise the IgE binding proteins of C. tala pollen. Skin prick tests using C. tala pollen extract were carried out in 25 patients with respiratory allergy. Celtis tala extract elicited positive skin reactions in five patients. Sensitised patients showed increased specific IgE levels to C. tala by immunodot. Immunoblotting assay demonstrated IgE binding to proteins of 50 kDa and 55 kDa. Celtis tala pollen extract is allergenically potent requiring approximately 50 ng of self-protein for 50% inhibition of IgE binding in enzyme_linked immunosorbent assay (ELISA) inhibition, whereas no cross-inhibition was detected with other types of pollen widely distributed in Argentina (Helianthus annuus and Prosopis spp.). Thus, these results taken together indicate that proteins of 50 kDa and 55 kDa may be considered as specific allergens of C. tala pollen.     

Allergenic components of Indian jujube (Zizyphus mauritiana) show IgE cross-reactivity with latex allergen

BACKGROUND: 'Latex-fruit syndrome' has been well documented. A prevalence of latex allergy among medical workers of 6.8-8.6% had been reported in Taiwan. However, there has been no study to determine the importance and type of fruit hypersensitivity in latex-allergic patients in Taiwan. This study aimed to identify the allergenic components of Indian jujube (Zizyphus mauritiana) and characterize the cross-reactivity of specific IgE antibodies to latex allergen. METHODS: Crude extracts were prepared from Indian jujube and from ammoniated natural rubber latex, and six medical workers and one patient with a history of fruit allergy underwent skin testing with routine allergens, latex, Indian jujube and other fruits. Sera from two Indian jujube skin test-positive latex-allergic subjects were used for allergen-specific IgE, immunoblotting, immunoblot inhibition and enzyme-linked immunosorbent assay (ELISA) inhibition studies. RESULTS: Both patients had positive skin test responses and specific IgE assays to Indian jujube and latex extracts. Immunoblotting revealed that IgE from both subjects bound to a 42-kD latex protein and a 42-kD Indian jujube protein. In addition, IgE from one subject bound to a 30-kD Indian jujube protein. Preincubation of atopic sera with Indian jujube or latex extract demonstrated absent and/or marked inhibition of IgE binding. Moreover, anti-Indian jujube protein antibody-based ELISA was able to detect latex extracts. CONCLUSIONS: Our results add to findings regarding the 'latex-fruit syndrome' described in the literature, and further study of the cross-reacting allergens identified in Indian jujube may help to elucidate the mechanisms underlying this syndrome.     

Identification of cross-reactive proteins amongst different Curvularia species

BACKGROUND: Curvularia lunata is an important inhalant allergen. The present study was undertaken to investigate the shared IgG- and IgE-binding components among seven Curvularia species prevalent in the aerospora. METHODS: Seven different Curvularia species were grown in a semisynthetic medium for 13 days. The extracts were analyzed by SDS-PAGE, immunoblot and ELISA/immunoblot inhibition using sera from C. lunata-positive patients and anti-C. lunata rabbit serum. RESULTS: Different Curvularia species showed 11-19 protein bands on SDS-PAGE. Proteins of 12, 20, 31, 45, 53, 78 and 97 kD were present in all the species. Eight out of 98 nasobronchial patients exhibited positive skin tests to C. lunata and to at least five Curvularia species. ELISA using these sera showed IgE binding with Curvularia species. Immunoblot using pooled anti-C. lunata sera from patients showed 5-12 allergenic proteins. Proteins of 12, 31, 45, 53 and 78 kD showed IgE binding in Curvularia species. Antibodies against C. lunata detected 6-14 antigenic proteins on immunoblot. Proteins of 31, 45 and 53 kD showed IgG binding in all the species. Proteins of 31 and 53 kD showed complete IgE/IgG binding inhibition. IgE/IgG ELISA inhibition showed dose-dependent inhibition in Curvularia species. C. lunata extract required 0.17 and 0.11 microg of protein for 50% IgE and IgG inhibition, respectively. C. clavata and C. pallescens required 10 times more protein to exhibit the same inhibition and other species required similar protein levels as those required by C. lunata. CONCLUSIONS: A high degree of cross-reactivity was observed between C. lunata and the six other Curvularia species tested. C. lunata and C. senegalensis shared maximum allergenic and antigenic components.