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1.
Deborah D Albright Diane Jordan-Wagner Diane C Napoli Amy L Parker Fonzie Quance-Fitch Bonnie Whisman Jacob W Collins Larry L Hagan 《Annals of allergy, asthma & immunology》2006,97(4):521-527
BACKGROUND: Multicolored Asian lady beetles (Harmonia axyridis) have been used as a biological control agent against crop-destroying aphids in the United States. Outside their natural habitat, H. axyridis seeks refuge in homes during fall and winter, leading to patient complaints and symptoms of rhinitis, wheezing, and urticaria on exposure to the beetles. OBJECTIVE: To gain a better understanding of the character and spectrum of allergic disease provoked by exposure to home-infesting lady beetles. METHODS: Eight patients with allergic symptoms suspected of being caused by H. axyridis and consistent with an IgE-mediated process were identified and interviewed. A whole-body extract from H. axyridis was prepared. Western blots using the patients' serum identified specific IgE antibodies in the extract. Through a novel technique, immunohistochemical analysis using beetle sections overlayed with patient serum was performed. A random survey of allergists from across the United States was also performed to evaluate experience with cases of lady beetle allergy. RESULTS: Western blots revealed IgE binding to 5 proteins with molecular weights of approximately 8.6, 21, 28, 31, and 75 kDa. Specific IgE bound to proteins localized in the beetle's mouth and leg areas. The allergist survey revealed positive responses in North Central, Mid-Atlantic and New England states. CONCLUSION: In 8 patients with allergic symptoms on exposure to high levels of lady beetles, specific IgE bound to proteins from H. axyridis. There was also an increased frequency of suspected cases of lady beetle allergy in endemic areas. 相似文献
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3.
Erwin EA Custis NJ Satinover SM Perzanowski MS Woodfolk JA Crane J Wickens K Platts-Mills TA 《The Journal of allergy and clinical immunology》2005,115(5):1029-1035
BACKGROUND: Commercially available assays for IgE antibody provide results in international units per milliliter for many allergen extracts, but this is not easily achieved with purified or novel allergens. OBJECTIVE: To develop assays for IgE antibody suitable for purified or novel allergens by using a commercially available immunosorbent. METHODS: Streptavidin coupled to a high-capacity immunosorbent (CAP) was used to bind biotinylated purified allergens from mite (Der p 1 and Der p 2), cat (Fel d 1), and dog (Can f 1). Assays for IgE antibody to these allergens were performed on sera from children (asthma and control) as well as adults with atopic dermatitis. RESULTS: The results were validated by serial dilution of sera with high and low levels of IgE antibody and were quantitated in international units per milliliter by using a standard curve. Values for IgE antibody to Der p 1, Der p 2, and Fel d 1 correlated with values obtained with the allergen extracts (r2 = 0.80, 0.84, and 0.95, respectively; P < .001 in each case). Furthermore, the values for IgE antibody in sera from children with high exposure to mite and cat allergens demonstrated 10-fold higher levels of IgE antibody to Der p 1 and Der p 2 than to Fel d 1 (P < .001). CONCLUSION: The streptavidin immunosorbent technique provides a new method for quantifying IgE antibody to purified proteins. The results provide evidence about the high quantities of IgE antibody to purified inhalant allergens in patients with atopic dermatitis. In addition, the results demonstrate major differences in IgE antibodies specific for mite and cat allergens among children with high exposure to both allergens. 相似文献
4.
Antigenic and structural analysis of group II allergens (Der f II and Der p II) from house dust mites (Dermatophagoides spp) 总被引:3,自引:0,他引:3
P W Heymann M D Chapman R C Aalberse J W Fox T A Platts-Mills 《The Journal of allergy and clinical immunology》1989,83(6):1055-1067
Monoclonal antibody affinity chromatography was used to purify two homologous mite allergens, Der f II from Dermatophagoides farinae and Der p II from D. pteronyssinus. They have the same molecular weight (MW) (15 kd) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, they have similar amino acid compositions, and their N-terminal amino acid sequences differ in only four of the first 35 residues. An excellent correlation was observed between IgE antibody to Der f II and Der p II measured in sera from 65 mite-allergic patients (r = 0.94; p less than 0.001) and between quantitative intradermal skin tests to both allergens. A third allergen (Der f III, MW 29 kd) was purified from D. farinae by repeated gel filtration. In sera from 51 mite-allergic patients, IgE antibody to Der f II, Der f III, and previously purified Der f I (MW 24 kd) was detected in 92%, 16%, and 78% of the sera by radioimmunoassay, respectively. Most patients, 41/51 (80%), demonstrated IgE antibody to more than one allergen. With monoclonal antibodies fully cross-reactive with Der f II and Der p II, a two-site immunoassay was developed for measuring absolute quantities (nanograms or micrograms) of these allergens. In extracts rich in mite-fecal material (n = 5), Der f I and Der p I (group I allergens) and Der f II and Der p II (group II allergens) were measured in ratios of 11:1 to 35:1. Lower ratios (1.1:1 to 7:1) were observed in mite body extracts (n = 6). These experiments clearly define a second group of major dust mite allergens that demonstrate extensive structural and antigenic homology. 相似文献
5.
Increased allergen production in turnip (Brassica rapa) by treatments activating defense mechanisms.
A R H?nninen J H Mikkola N Kalkkinen K Turjanmaa L Ylitalo T Reunala T Palosuo 《The Journal of allergy and clinical immunology》1999,104(1):194-201
BACKGROUND: Practical applications to enhance the productivity of agriculture by using plants with improved resistance to pathogens are expected to increase in the near future. Defense proteins play an important role in pathogen resistance, and some defense-related proteins are significant cross-reacting allergens. For example, cross-allergies are common among patients allergic to natural rubber latex (NRL), which contains many defense-related proteins. OBJECTIVE: Using a model plant (ie, turnip), we studied whether allergen contents increase after treatments activating defense mechanisms of the plants. METHODS: Whole or wounded turnips treated with salicylic acid, ethephon, or water were incubated for 2, 4, or 8 days. Allergen content was investigated by IgE immunoblotting with sera from patients allergic to NRL. An induced protein that bound IgE most intensively was purified and further characterized by mass analysis, amino acid sequencing, IgE-ELISA, and skin prick tests. RESULTS: In immunoblotting, clear IgE-binding bands were discernible only in samples from chemically treated plants. IgE was bound most intensively to a protein with an apparent molecular weight of 25 kd in SDS-PAGE and with a determined molecular weight of 18.7 kd. Sequenced peptides of the 18.7-kd protein showed over 70% homology to prohevein, a major allergen of NRL, and to many other prohevein-like defense proteins. In ELISA, sera from 30 of 34 (88%) adults and 21 of 26 (81%) children previously shown to have IgE against prohevein bound to the purified protein. In skin prick testing with the protein, 4 of 6 patients allergic to NRL had positive reactions. CONCLUSION: These results demonstrate that activating defense mechanisms of plants may considerably increase their allergen content. 相似文献
6.
Occupational asthma and rhinitis related to laboratory rats: serum IgG and IgE antibodies to the rat urinary allergen 总被引:6,自引:0,他引:6
T A Platts-Mills J Longbottom J Edwards A Cockroft S Wilkins 《The Journal of allergy and clinical immunology》1987,79(3):505-515
Allergic reactions to rat urinary proteins are an important cause of occupational asthma and rhinitis among laboratory workers. We have measured IgG and IgE antibodies to a purified rat urinary allergen in sera from 179 laboratory workers of whom 30 reported symptoms on exposure to rats. There was a very good correlation between IgE antibodies and positive skin tests. In addition, there was a close correlation between reported asthmatic reactions and serum IgE antibody to rat allergen: IgE ab was present in 12/18 of workers with asthmatic reactions but in only 2/135 of workers without symptoms (p less than 0.001). Serum IgG antibodies to rat allergen were present in all sera with IgE antibody but were also present in 30% of asymptomatic individuals. The incidence and quantity of IgG antibody correlated with the degree of exposure to animals (i.e., hours per day) but not with the length of exposure in years. Our results on rat allergy confirm that there is an increased incidence of asthma among individuals who were atopic as judged by positive skin tests to other allergens. However, this relationship did not apply to individuals with rhinitis alone, and excluding atopic individuals from employment would have been a very inefficient method of reducing asthma or rhinitis in this group. Our results confirm that IgE antibody responses to rat urinary allergen are an important cause of occupational disease. The results for IgG antibody suggest that their prevalence represents a marker for the degree of exposure to rat proteins. 相似文献
7.
Sánchez-Monge R Pascual CY Díaz-Perales A Fernández-Crespo J Martín-Esteban M Salcedo G 《The Journal of allergy and clinical immunology》2000,106(5):955-961
BACKGROUND: Lentils seem to be the most common legume implicated in pediatric allergic patients in the Mediterranean area. However, no lentil allergen has been isolated and characterized. OBJECTIVE: We sought to purify and characterize relevant IgE-binding proteins from boiled lentil extracts. METHODS: IgE-binding proteins from crude and boiled lentil extracts were detected with a pool of sera from patients with lentil allergy. Allergens were isolated by gel-filtration chromatography followed by cation- and anion-exchange chromatography or by reverse-phase HPLC. Their characterization included N-terminal amino acid sequencing, complex asparagine-linked glycan detection, specific IgE immunodetection with 22 individual sera from allergic patients, and immunoblot and CAP inhibition assays. RESULTS: Heat treatment of lentils produced substantial changes in the SDS-PAGE patterns of whole extracts, mainly a strong increase of 12- to 16-kd bands and a decrease of 25- to 45-kd components. Major IgE-binding proteins from the boiled lentil extract were located in the 12- to 16-kd and 45- to 70-kd ranges. Two allergens of 16 kd, proteins L1 and L2, and another one of 12 kd, protein L3, were purified. N-terminal sequencing indicated that all 3 were related and allowed their identification as gamma-vicilin subunits. Protein L1 was recognized by 68% of the individual sera tested and inhibited 64% of the IgE binding by commercial lentil CAPs. A second type of allergen of 66 kd, named protein H, was also isolated and identified as a seed-specific biotinylated protein. Protein H reacted with 41% of the individual sera and produced 45% inhibition in CAP inhibition assays. CONCLUSIONS: Two different types of allergens have been identified in boiled lentils. Those of 12 to 16 kd, called Len c 1, correspond to gamma-vicilin subunits, and those of 66 kd, designated Len c 2, correspond to seed-specific biotinylated protein. Homology with proteins from other legume species can explain potential cross-reactions among these foods. 相似文献
8.
Allergy to castor bean. II. Identification of the major allergens in castor bean seeds 总被引:6,自引:1,他引:6
S C Thorpe D M Kemeny R C Panzani B McGurl M Lord 《The Journal of allergy and clinical immunology》1988,82(1):67-72
Castor bean proteins were separated and identified by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted onto nitrocellulose paper. The capacity of the castor bean proteins to bind human IgE was probed with sera from castor bean-sensitive patients and radiolabeled anti-IgE. It proved difficult to identify allergens with isoelectric focusing. However, three allergens were identified when proteins were first separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis: the 2S storage albumin, the 11S crystalloid proteins, and a third protein doublet with molecular weights of 47 and 51 kd. Specific IgE antibody to the 2S storage albumin, measured by RAST, was detected in most (96%) castor bean-sensitive patients, confirming it as the major allergen. We would like to suggest that the 2S albumin be named Ric c I, that the crystalloid proteins be named Ric c II, and that the 47/51 kd doublet be named allergen 3. 相似文献
9.
BACKGROUND: We characterized by immunoblotting the antigenicity of the most frequent fungi colonizing cork during its industrial processing, Penicillium glabrum and Chrysonilia sitophila. Penicillium glabrum is the main causative agent of Suberosis, a hypersensitivity pneumonitis of cork workers. Chrysonilia sitophila induces both IgE sensitization and occupational asthma in the wood processing industry. METHODS: Serum-specific IgG, IgG4 and IgE to P. glabrum and C. sitophila from nine cork workers with hypersensitivity pneumonitis (HP) and seven with asthma (four with occupational asthma) were analysed by immunoblotting. RESULTS: Both HP and asthmatic patients' sera showed immunoreactivity to several proteins resolved in the specific immunoblot strips. The frequency of specific IgG recognition to 12-13.5 and 33 kDa proteins of P. glabrum was significantly higher in HP patients. The sera of HP patients had significantly higher specific IgG recognition to 16 and 51-55 kDa proteins of C. sitophila. There was no specific IgE recognition in the sera of HP or asthmatic patients to both fungi. CONCLUSIONS: Different patterns of antibody reactivity to P. glabrum and C. sitophila are seen in cork workers with hypersensitivity pneumonitis or asthma. The 12-13.5 and 33 kDa proteins of P. glabrum and the 16 and 51-55 kDa proteins of C. sitophila may be major antigens in Suberosis. 相似文献
10.
Yoshiko Asako Akemi Saito Hiroshi Yasueda Hiroshi Kawaguchi Kazuo Akiyama Masahiro Endo Yoshimi Onishi Kazutoh Takesako 《Arerugī》2002,51(8):615-621
We analyzed the reactivities of a series of purified allergens from Candida albicans (C. albicans) and Malassezia furfur (M. furfur) with IgE antibodies in sera from patients with atopic dermatitis. We compared the specific IgE antibody levels to manganese superoxide dismutase (Mn SOD), cyclophilin, enolase, secretory aspartic protease (SAP 2) and type A mannan from C. albicans and Mn SOD, cyclophilin and Mal f 2 from M. furfur in 21 sera from patients with atopic dermatitis and 20 sera from patients with asthma without atopic dermatitis. The prevalence of IgE antibodies and the mean IgE antibody levels to all of the allergens tested were higher among patients with atopic dermatitis than among those with asthma without atopic dermatitis. More than 50% of patients with atopic dermatitis were IgE antibody-positive to Mn SOD, cyclophilin and type A mannan from C. albicans, and Mn SOD and cyclophilin from M. furfur. The availability of these purified allergens will facilitate studies on the contribution of fungal allergens to the development of atopic dermatitis. 相似文献
11.
Suzanne S. Teuber MDa b W.Rich Peterson BSb 《The Journal of allergy and clinical immunology》1999,103(6):1180-1185
BACKGROUND: Two patients with tree nut allergy manifested by life-threatening systemic reactions reported the subsequent onset of systemic reactions after the consumption of coconut. OBJECTIVE: Herein, the IgE-binding proteins from coconut are described, and in vitro cross-reactivity with other nuts is investigated. METHODS: The IgE-binding profile of coconut endosperm tissue extract was analyzed by SDS-PAGE followed by immunoblotting. Immunoblot inhibition studies with walnut, almond, peanut, and coconut were performed. RESULTS: Sera IgE from both patients recognized reduced coconut allergens with molecular weights of 35 and 36.5 kd. IgE from 1 patient also bound a 55-kd antigen. Preabsorption of sera with nut extracts suppressed IgE binding to coconut proteins. Preabsorption of sera with coconut caused the disappearance of IgE binding to protein bands at 35 and 36 kd on a reduced immunoblot of walnut protein extract in 1 patient and suppression of IgE binding to a protein at 36 kd in the other patient. CONCLUSION: The reduced coconut protein at 35 kd was previously shown to be immunologically similar to soy glycinin (legumin group of seed storage proteins). The clinical reactivity in these 2 patients is likely due to cross-reacting IgE antibodies primarily directed against walnut, the original clinical allergy reported, and most likely to a walnut legumin-like protein. Coconut allergy in patients with tree nut allergy is rare; these are the first 2 patients ever reported, and therefore there is no general indication to advise patients with tree nut allergy to avoid coconut. 相似文献
12.
Satinover SM Reefer AJ Pomes A Chapman MD Platts-Mills TA Woodfolk JA 《The Journal of allergy and clinical immunology》2005,115(4):803-809
BACKGROUND: The specificity of serum antibody responses to different cockroach allergens has not been studied. OBJECTIVE: We sought to quantitate serum IgE and IgG antibodies to a panel of purified cockroach allergens among cockroach-sensitized subjects. METHODS: IgE antibodies to recombinant cockroach allergens (rBla g 1, rBla g 2, rBla g 4, rBla g 5, and rPer a 7) were measured in sera containing IgE antibodies to Blattella germanica extract (n = 118) by using a streptavidin CAP assay and a multiplex flow cytometric assay. Specific IgG antibodies were determined by using radioimmunoprecipitation techniques. RESULTS: Specific IgE antibodies measured by means of CAP assay and multiplex assay were strongly correlated ( r = 0.8, P < .001). The sum of IgE antibodies (in international units per milliliter) against all 5 allergens equated to IgE antibodies to cockroach extract. Although the prevalence of IgE antibodies was highest for rBla g 2 (54.4%) and rBla g 5 (37.4%), patterns of IgE antibody binding were unique to each subject. Surprisingly, only 16% of cockroach-sensitized subjects with IgE antibodies to house dust mite exhibited IgE antibody binding to cockroach tropomyosin (rPer a 7). Specific IgE antibodies were associated with increased IgG antibody levels, although detection of IgG in the absence of IgE was not uncommon. CONCLUSION: The techniques described offer a new approach for defining the hierarchy of purified allergens. IgE antibodies directed against 5 allergens constitute the majority of the IgE antibody repertoire for cockroach. Such distinct patterns of IgE-IgG responsiveness to different cockroach allergens highlight the complexity of B-cell responses to environmental allergens. 相似文献
13.
C Ebner T Birkner R Valenta H Rumpold M Breitenbach O Scheiner D Kraft 《The Journal of allergy and clinical immunology》1991,88(4):588-594
Eighty-three sera from patients with birch-pollen allergy were investigated for IgE antibodies against apple allergens by means of immunoblotting. In immunoblots, 81 patients (97.6%) exhibited IgE directed against the major allergen of birch, Bet v I (17 kd), and these patients also demonstrated IgE binding to apple allergens in the molecular weight range 17 to 18 kd. Inhibition studies by preincubation of sera with birch-pollen extract led to complete blocking of IgE binding to this 17 to 18 kd protein, whereas preincubation with apple extract could not diminish IgE binding to Bet V I. Furthermore, a 17 kd protein in apple extract could be detected by immunoblotting with a Bet v I-specific monoclonal antibody. Northern blotting with a Bet v I cDNA clone as a probe revealed cross-hybridization of birch and apple allergen coding nucleic acids under conditions of high stringency, suggesting significant homology of the nucleic acid level. Our results support the concept that antigens in birch pollen and apples share allergenic epitopes leading to IgE cross-reactivities that may cause clinical manifestations when a special threshold level of specific IgE antibodies is reached. 相似文献
14.
López-Torrejón G Salcedo G Martín-Esteban M Díaz-Perales A Pascual CY Sánchez-Monge R 《The Journal of allergy and clinical immunology》2003,112(6):1208-1215
BACKGROUND: Lentils are among the main plant foods causing allergic reactions in pediatric patients in the Mediterranean area and in many Asian communities. However, very few reports have been devoted to identifying lentil allergens. Seed storage proteins of the vicilin family have been characterized as major allergens in several seed legumes and tree nuts. OBJECTIVE: We sought to evaluate the role of lentil vicilins as food allergens. METHODS: A serum pool and individual sera from 22 patients with lentil allergy were used in different IgE-binding assays. Mature lentil vicilin was isolated by means of cation-exchange chromatography, followed by reverse-phase HPLC, and characterized by means of N-terminal amino acid sequencing, matrix-assisted laser desorption/ionization mass spectrometry (MALDI) analysis, complex asparagine-linked glycan detection, specific IgE immunodetection with individual sera, and ELISA inhibition assays. Complete cDNAs encoding lentil vicilin variants were isolated by means of PCR with primers based on the amino acid sequence of the allergen. RESULTS: A major IgE-binding component of approximately 50 kd was detected in lentil extracts. This component was isolated and characterized, showing a single N-terminal amino acid sequence homologous to those of legume vicilins and a broad peak (maximum at 48613 d) in MALDI analysis. The purified allergen was recognized by 77% (17/22) of the individual sera from patients with lentil allergy and reached up to 65% inhibition of the IgE binding to the crude lentil extract. The allergen showed 3 isoforms varying in their degree of N-glycosylation. Two cDNA clones encoding different allergen variants were isolated. The amino acid sequences deduced from both clones (415 and 418 residues; 47.4 and 47.8 kd) showed greater than 50% identity with major peanut (Ara h 1) and soybean (conglutinin subunits) allergens belonging to the vicilin family. Furthermore, these sequences included those of the previously characterized lentil allergen Len c 1.02 (108 amino acid residues of the C-terminal domain) and those of a novel lentil IgE-binding protein of 26 kd. CONCLUSION: The mature 48-kd lentil vicilin, designated Len c 1.01, is a major allergen. Two of its processing fragments, corresponding to subunits of 12 to 16 kd (previously named Len c 1) and 26 kd, are also relevant lentil IgE-binding proteins. The sequence homology of Len c 1.01 to those of major allergens from peanut, soybean, walnut, and cashew can help to investigate potential cross-reactions among these plant foods. 相似文献
15.
Identification and partial characterization of the soybean-dust allergens involved in the Barcelona asthma epidemic 总被引:6,自引:0,他引:6
M J Rodrigo F Morell R M Helm M Swanson A Greife J M Anto J Sunyer C E Reed 《The Journal of allergy and clinical immunology》1990,85(4):778-784
Asthma epidemics in Barcelona, Spain, have been attributed to dust generated by the unloading of soybeans in the harbor. Sera of four different groups of 10 subjects in each group were studied: (1) patients attending an emergency room in Barcelona for an asthma attack on epidemic days, group A, (2) patients attending an emergency room for an attack on nonepidemic days, group B, (3) patients with asthma from other cities, group C, and (4) patients without asthma from Barcelona matched by age and sex with group A, group D. All subjects in group A had IgE to allergens in extracts of various soybean samples. In contrast, only one of the 10 subjects in each of groups B and C and none of those subjects in group D had IgE to uncleaned bean and hull extracts. Radioimmunoassay demonstrated that in sera from patients with asthma during an asthma outbreak reacted primarily to soybean hull and dust extracts. Sodium dodecyl sulfate-polyacrylamide and gel electrophoresis thin-layer isoelectrofocusing demonstrated protein bands of 97.4 to less than 14.4 kd and isoelectric point between 6 and 3.5. By Western blot and thin layer isoelectrofocusing/blotted radioimmunoisoelectrofocusing, IgE of patients with asthma during an asthma outbreak reacted weakly to two protein bands of molecular weight ranging from 42 to 21 kd, strongly to glycoprotein bands with molecular weight less than 14.4 kd, and isoelectric point less than 6, which appeared to be the major allergens. 相似文献
16.
J. PEPYS W. E. PARISH B. STENIUS-AARNIALA† L. WIDE‡ 《Clinical and experimental allergy》1979,9(6):645-658
The sera of atopic and non-atopic persons with allergic pulmonary disorders were examined for long-term sensitizing, IgE and short-term sensitizing heat-stable (S-TS) antibodies which were present separately or together in the sera of some patients sensitive to antigens such as budgerigar serum proteins and Aspergillus fumigatus. In fourteen atopic patients with extrinsic asthma, six had both types of antibody to common allergens, and of nine non-atopic patients with cryptogenic (intrinsic) asthma, four had only heat-stable short-term sensitizing antibodies. The sera of atopic subjects with type I prick test reactions and positive RAST's, showed specific IgE antibody by baboon PCA tests to budgerigar serum proteins, A. fumigatus, Timothy grass pollen extract and hen egg extract, and not to Dermatophagoides farinae, possibly because of naturally occurring mite antibodies in the baboon. The sera of non-atopic asthmatics, who had given negative prick test but positive immediate, dual or late intracutaneous tests, and only late asthmatic reactions, contained precipitins in most cases and gave little or no RAST reaction. On baboon PCA these sera contained either, S-TS antibody alone, or S-TS plus long-term sensitizing antibody, or long-term sensitizing antibody alone. Some of the sera with long-term sensitizing antibody contained blocking antibody which could diffuse away in the 24 hr delay for the baboon PCA test and could also be responsible for the negative RAST. Tests with insoluble anti-IgE immuno-adsorbents on two sera from persons sensitive to aspergillus confirmed that the S-TS activity was not due to IgE, and on two sera with negative RAST and negative prick tests to budgerigar serum antigens confirmed that the 24 hr monkey PCA responses were due to IgE. 相似文献
17.
Aqueous extracts of white oak pollen were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes. The nitrocellulose membranes were blocked with phosphate-buffered saline 15% nonfat dry milk, incubated with dilutions of sera from atopic or control subjects, and probed with a radiolabeled or peroxidase-labeled antihuman IgE. The IgE binding bands were detected by autoradiography or enzymatic reaction; 45 to 50 protein bands were observed in silver-stained gels. IgE from 30 of the 38 sera tested from oak-sensitive subjects bound to 23 bands with molecular weights (MWs) between 106 to 108 kd (band 1) and 13.2 to 15.2 kd (band 23). No band was recognized by sera of every patient. Band 5 (MW 74.0 to 77.9 kd) and band 21 (MW 16.2 to 17.7 kd) were recognized by 71% of the patients' sera. Multiple bands were recognized by 30% to 50% of the sera tested. All patients who were skin test positive to oak by prick testing had positive immunoblots. Of 12 patients positive by intradermal skin testing, only four patients had positive immunoblots. The average number of allergens recognized by a single patient was 6.6. The maximum number of allergens to which any individual reacted was 18; the minimum number was one. Extracts separated under nonreducing conditions resulted in aggregates that did not enter the polyacrylamide gel. Of the protein that did enter the gel, the higher MW species elicited banding patterns similar to patterns observed under reducing conditions, whereas lower MW IgE binding bands were lost. These data suggest that the extractable proteins of white oak pollen contain multiple proteins that are potentially allergenic. 相似文献
18.
Kurup Banerjee Hemmann Greenberger Blaser & Crameri 《Clinical and experimental allergy》2000,30(7):988-993
BACKGROUND: Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity lung disease resulting from exposure to Aspergillus fumigatus allergens. Patients with ABPA show elevated Aspergillus-specific serum IgE, a major criterion used in the diagnosis of the disease. Crude culture filtrate and mycelial antigens have been used widely to demonstrate IgE antibody to Aspergillus in the sera of patients. While these antigens have been useful in the diagnosis of ABPA, occasionally they present inconsistency in their reactivity and lack of specificity. Although in recent years, a number of purified A. fumigatus allergens have been produced by molecular cloning, no attempt was made to evaluate them systematically. OBJECTIVE: To evaluate the recombinant proteins from A. fumigatus for their IgE antibody binding, we studied sera from ABPA patients and controls by antigen specific enzyme linked immunosorbent assay (ELISA). METHODS: Recombinant Aspergillus allergens Asp f 1, f 2, f 3, f 4, and f 6 were studied for their specific binding to IgE in the sera of ABPA patients, A. fumigatus skin prick test positive asthmatics, and normal controls from the USA and Switzerland. The sera were blinded and studied by ELISA in two different laboratories. RESULTS: All the recombinant allergens showed IgE antibody binding with sera from patients with ABPA, whereas only fewer asthmatics and normal sera showed significant binding. The three selected recombinant allergens together reacted with all the ABPA patients studied. CONCLUSIONS: The results demonstrate that Asp f 2, f 4, and f 6 can be used in the serodiagnosis of ABPA, while IgE antibody binding to Asp f 1 and f 3 was not specific. 相似文献
19.
Run-Yee Lin MD Horng-Der Shen MS Shou-Hwa Han MD PhD 《The Journal of allergy and clinical immunology》1993,92(6)
The allergenic components of the shrimp (Parapenaeus fissurus) were identified by immunoblotting, with sera from 10 allergic patients. Six components, ranging in molecular weight from about 86 to 39 kd, showed IgE-binding activity and were identified as allergens of the shrimp. The component with a molecular weight of about 39 kd showed the highest frequency of IgE binding (70%) and was considered to be one of its major allergens. Two monoclonal antibodies against this 39 kd component were generated, and their antigenic cross-reactivity with five different kinds of seafood, shrimp, crab, cuttlefish, oyster, and pomfret was analyzed. Monoclonal antibody 1-6-10B reacted with the 39 kd component from shrimp only, but monoclonal antibody 2-7-IE also reacted with the 39 kd component from crab. By extraction with 0.5% sodium dodecylsulfate and ethanol precipitation, a highly purified shrimp 39 kd component was obtained. In two-dimensional gel electrophoresis six isoforms of this purified 39 kd component, with isoelectric point values from purified 39 kd component, with isoloelectric point values from 5.1 to 5.6, were identified. No marked difference was observed when the amino acid composition of this purified 39 kd allergen was compared with those of serum albumin from different animals. They all contain a high proportion of acidic amino acids. There was also a 62% to 83% sequence homology among three different pairs of peptide fragments of purified 39 kd components of shrimp and crab. In conclusion, a 39 kd major allergen from the shrimp has been identified and characterized in the present study. According to the suggestions of the International Union of Immunological Societies, this allergen is designated as Par f I. 相似文献
20.
Belén Añibarro Jose L. Fontela MD Francisco De La Hoz PhD 《The Journal of allergy and clinical immunology》1997,100(6):734-738
Background: Garlic dust has not been a frequently encountered cause of IgE-mediated disease. Objective: We report on 12 patients (all of them garlic workers) with the clinical criteria for occupational asthma. Methods: Skin prick tests and serum-specific IgE determinations were performed with common inhalants, garlic, and other members of the Liliaceae family (onion, leek, and asparagus). Bronchial challenge test with garlic powder was performed in all patients. Garlic and onion extract proteins were separated by sodium dodecylsulfate–polyacrylamide gel electrophoresis. Immunoblot and IgE immunoblot inhibition analyses were performed with patients' sera on extracts of garlic, onion, and pollens of Phleum pratense and Chenopodium album. Results: Garlic sensitization was demonstrated by bronchial challenge test in seven patients (group 1) and ruled out in the remaining five (group 2). Clinical data were similar in both groups. The patients with garlic allergy had a mean age of 27 years, and all of them had pollen allergy; sensitization to other members of the Liliaceae family was also common. Electrophoresis of garlic extract revealed two major protein bands at approximately 12 and 54 kd. During IgE immunoblotting, the pool of sera reacted with garlic proteins mainly at 54 kd. Preincubation with onion, Phleum, and Chenopodium partially abolished the IgE binding to several allergens of garlic. Conclusion: We report on seven patients in whom an occupational garlic allergy was demonstrated. Garlic allergy is relatively rare but seems to affect young subjects with pollen allergy, and sensitization to other members of the Liliaceae family is common. The results of this study confirm the presence of some structurally similar allergens in garlic, onion, and certain pollens. (J Allergy Clin Immunol 1997;100:734-8.) 相似文献