Typical levels of airborne fungal spores in houses without obvious moisture problems during a rainy season in Florida, USA.

OBJECTIVE: The aim of this study was to determine types and levels of airborne fungal spores in air-conditioned homes built after 1980 without obvious moisture problems during the 2004 summer (rainy season) in central Florida, USA. METHODS: Eighteen single-family homes were selected based on protocol questionnaire and cursory inspection, which revealed no obvious moisture or visible fungal growth. Non-cultured spores were collected with Air-O-Cell cassettes. Three indoor air samples and 2 outdoor air samples were collected from each home. One indoor and 2 outdoor samples were not interpretable. Fifty-three indoor and 34 outdoor air samples were analyzed by optical microscopy. RESULTS: Several spore types were detected in the indoor samples, at levels generally lower than those detected in the outdoor samples. Spores from the Penicillium/Aspergillus group were the most prevalent types indoors, exceeding the absolute levels and relative percentages of these spores outdoors. Ascospores and basidiospores were the most prevalent spore types outdoors. The percentages of other spore types (Cladosporium and Curvularia) were similar in the indoor and outdoor samples. Moisture-indicator fungi (Chaetomium, Stachybotrys, and Ulocladium species) were nearly absent in both indoor and outdoor samples. CONCLUSION: Airborne fungal spores are present in average central Florida homes without obvious moisture problems during the summer, at levels that are lower than those found outdoors. Spores from the Penicillium/Aspergillus group are prevalent in these homes, and moisture-indicator fungi (Chaetomium, Stachybotrys, and Ulocladium species) are nearly absent. Despite climatic differences, airborne fungal spore types and levels in central Florida houses are similar to those found in other geographical locations.

Allergenicity of varieties of soybean.

BACKGROUND: Soybean hulls (SHs) cause respiratory allergies. This study investigates the allergenicity of soybean varieties (SVs) by in vivo and in vitro tests. METHODS: Ten SVs were studied: (a) five with a proved clinical relevance (SVs 1, 2, 3, 4, 5), the last four with a 'dull' phenotype; (b) five of undetermined relevance, three of them (SVs 6, 7, 8) with a 'shiny' phenotype, and two (SVs 9 and 10) with a 'dull' phenotype. Extracts from all 10 SVs were used to skin prick test (SPT) 21 subjects sensitized to SHs. Positive and negative sera pools prepared from sera of subjects sensitized or not to SHs, respectively, were utilized to perform in vitro experiments (specific IgE and IgG4 determinations, SDS-PAGE/IgE-Western blot, and IgE-inhibition). RESULTS: In this study, it was found that 52.4, 52.4, 57.1, 71.4, 80.9, 42.9, 57.1, 71.4, 52.4, and 38.1% subjects had a positive SPT with SVs 1-10, respectively (P NS). Specific IgE values to SVs 1-10 obtained with the positive pool are 28.3, 26.4, 29.9, 28.3, 26.8, 4.8, 13.4, 6.7, 24.7, and 17.5% total counts bound, respectively; and specific IgG4 values 0.851, 0.818, 0.721, 1.609, 0.789, 0.617, 0.662, 0.0, 1.127, and 0.934 OD units, respectively; the microgram of protein required to produce 50% inhibition are 2.5, 3.7, 4.5, 2.4, 5, 39.8, 25.2, 25.1, 4.5, and 8.9, respectively. A 7-kDa band is present in all SVs except in those with a 'shiny' phenotype. CONCLUSIONS: The SVs with a 'shiny' phenotype contain less allergens than the other SVs studied, as determined by in vitro tests. However, SPT results with the SVs do not differ. Genetic screens should be devised to select plants with reduced, preferably absent, allergenicity, but with a high nutritional value, and this allergenicity should be studied utilizing in vivo and in vitro tests.

Disodium octaborate tetrahydrate (DOT) application and vacuum cleaning, a combined strategy to control house dust mites.

BACKGROUND: The effectiveness of acaricides in homes is controversial. OBJECTIVE: To determine whether disodium octaborate tetrahydrate (DOT) combined with vacuuming lowers dust mite numbers and their allergens in carpets and sofas. METHODS: A 6-month study was carried out with 93 homes, which were randomized into three groups: (i). active, received DOT; (ii). placebo, received water; and (iii). control, received no application. Active and placebo homes were vacuumed weekly. Dust was collected from carpets and sofas at the start of the study and every 2 months thereafter and quantified for live, total mites, and mite allergen levels. RESULTS: At 2 months, live mite numbers in active carpets were 3 +/- 1, in placebo carpets 129 +/- 48, and in control carpets 177 +/- 39 mites/g. The corresponding numbers in sofas were 3 +/- 2, 81 +/- 31, and 134 +/- 45 mites/g, respectively (P < 0.001 active vs placebo and vs. control). Live mites in carpets and sofas remained lower in the active group at 6 months (P < 0.001). Total mites in active carpets decreased from 555 +/- 69 at baseline to 223 +/- 32 mites/g at 6 months (P < 0.001) and mite allergen levels from 1.36 +/- 0.13 to 0.85 +/- 0.16 microg/g (P < 0.001). Total mites in active sofas remained unchanged, but mite allergen levels decreased from 1.48 +/- 0.25 at baseline to 0.7 +/- 0.15 microg/g at month 6 (P < 0.05). CONCLUSION: DOT kills mites in carpets and sofas, and, combined with vacuuming, effectively reduces total mites in carpets and mite allergen levels in carpets and sofas.

Identification of the soybean hull allergens involved in sensitization to soybean dust in a rural population from Argentina and N-terminal sequence of a major 50 KD allergen.

BACKGROUND: Sensitization to soybean hull (SH) allergens occurs in subjects from Argentina, a soybean producer country. However, the causative allergens have not been identified. The purposes of this study are to: (i) identify the SH allergens using sera of 29 subjects with asthma and/or allergic rhinitis from Argentina exposed to soybean dust who have a positive (weal with SH/weal with histamine > or = 0.5) skin prick test to SH; and (ii) determine the N-terminal amino acid sequence of a major 50 K SH allergen that sensitizes this population. METHODS: All sera were assayed for specific IgE (RIA), IgG4 (ELISA), and IgE and IgG4-Western blots. A sera pool from 10 healthy subjects was a negative control. N-terminal amino acid sequencing was performed by the Edman degradation method. RESULTS: Positive specific IgE only was found in 12/29 (41.4%), IgG4 in 3/29 (10.3%), and both IgE and IgG4 in 14/29 (48.3%) sera. IgE-Western blot demonstrates: (i) an allergen, MW 50 K (51.7% binding); (ii) one or two distinct allergens, MW < 20.2 K (72.4% binding), depending on the sera; and (iii) 1-5 additional IgE binding proteins, MW > 20.2 to < 46.9 K (41.4% binding), depending on the sera. IgG4-Western blot demonstrates: (i) a band, MW 70K (31% binding); (ii) a band, MW 50 K (17.2% binding); (iii) one or two additional bands, MW < 20.2 K (51.7% binding), depending on the sera; and (iv) a band, MW > 20.2 to < 28.5 K (20.7% binding). The 50 K allergen N-terminal amino acid sequence of the first 17 amino acids indicates a significant homology with chlorophyll A-B binding protein precursors from tomato, spinach, and petunia. CONCLUSIONS: Specific IgE and IgG4 to SH are common in sera from allergic individuals living in rural areas in Argentina. SH contain an IgE binding protein, MW about 50 K, not previously described. Sensitization to this allergen is common in subjects who are repeatedly exposed to soybean dust inhalation.

House dust mite, cat, and cockroach allergen concentrations in daycare centers in Tampa, Florida.

BACKGROUND: Allergen exposure in early childhood is a risk factor for sensitization and the development of asthma. Studies performed in Europe, New Zealand, and Singapore indicated the presence of indoor allergens in childcare centers and schools. However, the importance of indoor allergens in daycare centers in humid and warm regions of the world is not known. OBJECTIVE: To measure total mite counts, Der p 1, Der f 1, Fel d 1, and Per a 1 allergens in dust samples and mite allergen airborne concentrations in daycare centers in Tampa, Florida, United States. METHODS: Twenty daycare centers were surveyed for mite, cat, and cockroach allergens in Tampa, FL. One dust and two air samples (one during the day and one during the night) were collected in each center. Dust samples were extracted and analyzed for mite (Der p 1 and Der f 1), cat (Fel d 1), and cockroach (Per a 1) allergens. Mite airborne allergen concentrations were analyzed by RAST inhibition and expressed in standardized mite allergen units per m3 of air (AU/m3). RESULTS: Mites were identified in 15 samples, and concentrations ranged from 10 to 1,200 mites/g (298 +/- 355.2). The most prevalent mite species was Dermatophagoides pteronyssinus ( Der p 1). Der p 1 and/or Der f 1 were detected in 10 daycare centers. Der p 1 was detected in eight centers and ranged from I to 21.8 microg/g of dust (5.4 +/- 6.9); Der f 1 was detected in 3 centers and ranged from 0.2 to 2.1 microg/g of dust (1.3 +/- 0.9). Per a 1 and Fel d 1 were detected in all centers in small quantities; Per a 1 ranged from 8 to 1,806 ng/g (263.1 +/- 449.7) and Fel d 1 from 0.2 to 120 U/g of dust (16.6 +/- 31.7), respectively. Airborne mite allergen was detected in 18 centers and ranged from 0.01 to 2.7 AU/m3 during the day (0.2 +/- 0.6) and from 0.01 to 0.12 AU/m3 during the night (0.06 +/- 0.03), P = 0.001. CONCLUSIONS: Mite, cat, and cockroach allergens are present in daycare centers in Tampa, FL. Mite allergen concentrations exceeded levels that have been associated with sensitization and symptoms in allergic subjects in 40% of these centers.

[Association between soybean dust exposure, allergic sensitivity and profile of respiratory symptoms]. / Asociación entre exposición a polvo de soja, sensibilidad alérgica y perfil de síntomas respiratorios.

The purpose of this study was to correlate soybean dust (SD) exposure, skin reactivity to soybean hull (SH) allergens, and symptoms of asthma and/or allergic rhinitis. A group of 365 subjects with asthma and/or allergic rhinitis and a control group of 50 individuals without respiratory symptoms were studied. The level of exposure to SD is defined as follows: 1) direct (DE); 2) indirect (ID), and 3) urban (UE). All subjects completed standard questionnaires. Skin tests with a SH extract and with common allergens were performed by the prick technique (SPT). Fifty-six (15.3%) patients and no subjects from control group had positive SPT (histamine index > or = 0.5) with a SH allergen extract. The percentages of positive SPT to SH extract were 38.7%, 20.3% and 8.4% in subjects with DE, IE and UE, respectively (p < 0.001). Monosensitization to SH was absent in all subjects. The percent of subjects with positive SPTs to mites (p < 0.01), pollen (p < 0.01) and molds (p < 0.05) were higher in subjects with a positive SPT to SH versus those with a negative SPT to SH. Sixty-six percent of subjects with DE and 13.6% of subjects with IE or UE reported respiratory symptoms after SD inhalation (Odds Ratio: 12.67 [2.4-74.9], p < 0.001). Compared to subjects exclusively sensitized to mites, patients sensitized to SH presented significantly different clinical characteristics. Soybean production has been increasing in Argentina during the last 20 years, determining an increase in the population exposed to chronic SD inhalation. This fact determines a high risk of sensitization and triggering of respiratory symptoms in atopic subjects. This study demonstrates that there is: 1) a high prevalence of skin reactivity to SH in subjects with asthma and/or allergic rhinitis from Argentina and that this prevalence is associated with the level of exposure to SD, and 2) an association between sensitivity to SH and severity of asthma. Measures to avoid release and inhalation of SD in rural areas from Argentina are needed.

Asociación entre exposición a polvo de soja, sensibilidad alérgica y perfil de síntomas respiratorios. / [Association between soybean dust exposure, allergic sensitivity and profile of respiratory symptoms]

The purpose of this study was to correlate soybean dust (SD) exposure, skin reactivity to soybean hull (SH) allergens, and symptoms of asthma and/or allergic rhinitis. A group of 365 subjects with asthma and/or allergic rhinitis and a control group of 50 individuals without respiratory symptoms were studied. The level of exposure to SD is defined as follows: 1) direct (DE); 2) indirect (ID), and 3) urban (UE). All subjects completed standard questionnaires. Skin tests with a SH extract and with common allergens were performed by the prick technique (SPT). Fifty-six (15.3

) patients and no subjects from control group had positive SPT (histamine index > or = 0.5) with a SH allergen extract. The percentages of positive SPT to SH extract were 38.7

, 20.3

and 8.4

in subjects with DE, IE and UE, respectively (p < 0.001). Monosensitization to SH was absent in all subjects. The percent of subjects with positive SPTs to mites (p < 0.01), pollen (p < 0.01) and molds (p < 0.05) were higher in subjects with a positive SPT to SH versus those with a negative SPT to SH. Sixty-six percent of subjects with DE and 13.6

of subjects with IE or UE reported respiratory symptoms after SD inhalation (Odds Ratio: 12.67 [2.4-74.9], p < 0.001). Compared to subjects exclusively sensitized to mites, patients sensitized to SH presented significantly different clinical characteristics. Soybean production has been increasing in Argentina during the last 20 years, determining an increase in the population exposed to chronic SD inhalation. This fact determines a high risk of sensitization and triggering of respiratory symptoms in atopic subjects. This study demonstrates that there is: 1) a high prevalence of skin reactivity to SH in subjects with asthma and/or allergic rhinitis from Argentina and that this prevalence is associated with the level of exposure to SD, and 2) an association between sensitivity to SH and severity of asthma. Measures to avoid release and inhalation of SD in rural areas from Argentina are needed.

Asociación entre exposición a polvo de soja, sensibilidad alérgica y perfil de síntomas respiratorios / [Association between soybean dust exposure, allergic sensitivity and profile of respiratory symptoms]

The purpose of this study was to correlate soybean dust (SD) exposure, skin reactivity to soybean hull (SH) allergens, and symptoms of asthma and/or allergic rhinitis. A group of 365 subjects with asthma and/or allergic rhinitis and a control group of 50 individuals without respiratory symptoms were studied. The level of exposure to SD is defined as follows: 1) direct (DE); 2) indirect (ID), and 3) urban (UE). All subjects completed standard questionnaires. Skin tests with a SH extract and with common allergens were performed by the prick technique (SPT). Fifty-six (15.3

) patients and no subjects from control group had positive SPT (histamine index > or = 0.5) with a SH allergen extract. The percentages of positive SPT to SH extract were 38.7

in subjects with DE, IE and UE, respectively (p < 0.001). Monosensitization to SH was absent in all subjects. The percent of subjects with positive SPTs to mites (p < 0.01), pollen (p < 0.01) and molds (p < 0.05) were higher in subjects with a positive SPT to SH versus those with a negative SPT to SH. Sixty-six percent of subjects with DE and 13.6

of subjects with IE or UE reported respiratory symptoms after SD inhalation (Odds Ratio: 12.67 [2.4-74.9], p < 0.001). Compared to subjects exclusively sensitized to mites, patients sensitized to SH presented significantly different clinical characteristics. Soybean production has been increasing in Argentina during the last 20 years, determining an increase in the population exposed to chronic SD inhalation. This fact determines a high risk of sensitization and triggering of respiratory symptoms in atopic subjects. This study demonstrates that there is: 1) a high prevalence of skin reactivity to SH in subjects with asthma and/or allergic rhinitis from Argentina and that this prevalence is associated with the level of exposure to SD, and 2) an association between sensitivity to SH and severity of asthma. Measures to avoid release and inhalation of SD in rural areas from Argentina are needed.

Sensitization to soybean hull allergens in subjects exposed to different levels of soybean dust inhalation in Argentina.

BACKGROUND: Soybean hulls (SHs) have been identified as the source of aeroallergens responsible for soybean asthma outbreaks. However, the prevalence of sensitization to SH allergens in subjects from Argentina, a country where soybeans are produced, is unknown. OBJECTIVE: The purpose of this study was to determine the prevalence of sensitization to SH by in vivo and in vitro tests in subjects with asthma or allergic rhinitis and in control subjects from Argentina who have been exposed to different levels of soybean dust inhalation (SDI). METHODS: Exposure to SDI is defined as follows: (1) direct = occupational, (2) indirect = proximity to soybean fields or grain elevators, and (3) urban = urbanized areas without a known source of SDI. Two groups were studied. Group 1 consisted of 365 subjects with asthma or allergic rhinitis and group 2 (control group) of 50 healthy individuals. Subjects from both groups were classified according to their exposure to SDI. All subjects completed standard questionnaires. Prick skin tests (STs) with an SH extract and with common allergens were performed on all subjects. Specific IgE and IgG4 to SH were measured in sera of 51 of 56 subjects from group 1 who had a positive ST to SH and in all sera from group 2. RESULTS: Fifty-six (15.3%) subjects from group 1 and no subjects from group 2 had a positive ST to SH (wheal SH/wheal histamine >/=0.5). In group 1, positive STs to SH were 38.7%, 20.3%, and 8.2% in subjects with direct, indirect, and urban exposures, respectively (P <.001). Monosensitization to SH is absent in all subjects from group 1. The percent of subjects with positive STs to mites, pollen, and molds was highest in those with a positive ST to SH versus those with a negative ST to SH (P <.01). Asthmatic patients with a positive ST to SH, compared with those exclusively sensitized to mites, had a higher frequency of daily or weekly symptoms (59.4% vs 25.7%, respectively, P <.001) and a higher percent of glucocorticoid dependence (52.8% vs 34%, respectively, P <.01). Percent positive IgE in group 1 and group 2 were 39.2% and 10% (P <.001) and percents positive IgG4 are 27.4% and 12%, respectively (not significant). In subjects from group 1 and group 2 with direct exposure percents positive IgE are 58.3% and 13.3% (P < .001) and percents positive IgG4 were 75% and 20%, respectively (P < .02). IgG4 in group 1 was significantly higher in subjects with direct exposure compared with subjects with indirect or urban exposure. CONCLUSION: This study demonstrated that there was (1) a high prevalence of sensitivity to SH in subjects with asthma or allergic rhinitis from Argentina and (2) an association between sensitivity to SH and severity of asthma and level of exposure to SDI.

Identification of the soybean hull allergens responsible for the Barcelona asthma outbreaks.

BACKGROUND: Soybean hulls were identified as the etiologic agent responsible for the asthma outbreaks that occurred in Barcelona and Cartagena, Spain. OBJECTIVE: To identify the main soybean hull allergens using the sera of 18 asthmatic epidemic patients from Barcelona and to compare the results to those previously reported. METHODS: Specific IgE to a soybean hull allergen extract was determined by radioimmunoassay and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in a 10-20% Tris-tricine gradient gel followed by Western blot. RESULTS: Specific IgE, which was detected in 15 of the 18 (83.3%) sera, indicated two nonoverlapping band patterns on the SDS-PAGE/Western blot: (1) pattern 1, present in 11 sera, consisted of three bands with estimated molecular weights (MWs) of 8, 7.5 and 7 kD, which are the MWs described for the allergens Gly m 2, Gly m 1A and Gly m 1B, respectively, and (2) pattern 2, present in 3 sera, consisted of a band with an estimated MW of 8.2-8.3 kD and four additional bands in a MW range of 25-36 kD. The remaining positive sera indicated very faint bands. CONCLUSIONS: This study confirms that soybean hulls contain three main allergens with MWs of 8, 7.5 and 7 kD, and indicates additional higher MW allergens, which selectively bind specific IgE of the sera that do not react with the three low MW components. This dichotomous and nonoverlapping pattern of allergen recognition has not been previously described.

Increased positivity of skin test and allergenic stability of glycerinated soybean hull extracts.

BACKGROUND: Results obtained from previous experiences with soybean hull antigens suggest that the addition of glycerine to the extract increases positivity of the skin test. OBJECTIVE: The purpose of this study was to investigate the effect of this glycerine addition and the influence of different storage temperatures on the potency of soybean hull extracts. METHODS: Twenty-two asthmatic patients admitted to emergency rooms during one of the soybean-related Barcelona asthma epidemics were evaluated 2 years after the last epidemic for: (1) sensitivity to prick test with glycerinated and non-glycerinated extracts, both fresh and stored at 4 degrees C, - 20 degrees C and - 70 degrees C for 30 days, and (2) specific IgE to a fresh hull extract. All extracts were also studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: No differences in positivities were detected for prick tests performed the same day of preparation between non-glycerinated (5/22; 22.7%) and glycerinated (4/22; 18.2%) extracts. After 30 days storage, positivities of prick test with non-glycerinated extracts stored at 4 degrees C did not significantly differ from the results obtained on the first day. However, prick tests performed with glycerinated extracts stored for 30 days showed increased positivity for the extracts stored at 4 degrees C (7/22; 31%) (P < 0.05) as well as for those stored at - 20 degrees C and - 70 degrees C (9/22; 40.9%) (P < 0.001). This latter percentage is similar to that of specific immunoglobulin E detected by radioimmunoassay (10/22; 45. 4%). Also, for the glycerinated extracts only, the quantitative response showed an increase in mean diameter of the induration at 30 days at the three temperatures, even though owing to the great standard deviation and limited number of patients, it was only significant for the extract stored at - 20 degrees C (P < 0.01). SDS-PAGE gel densitometry at 30 days demonstrated a loss of protein in the bands with a molecular weight higher than 66.2 kDa for non-glycerinated extracts stored at 4 degrees C. CONCLUSIONS: The results of this study confirm previous observations with soybean hull allergen extracts and indicate that (1) storage of soybean hull extracts with glycerine addition for 30 days at the concentration studied (1:100 w/v) determines a significant increase in the positivity of skin tests, and (2) glycerine addition stabilizes soybean hull extracts at any of the temperatures studied.

Neoallergens in heated soybean hull.

BACKGROUND: During the process of harvest, transport and storage, microbial and mold contamination can raise the temperature of soybeans to 75 degreesC or higher. The purposes of this study were (1) to evaluate the allergenicity of fresh and stored soybean hulls and (2) to ascertain whether heat alters the allergenicity of stored soybean hulls. METHODS: Allergen extracts were prepared from (1) stored soybean hulls, (2) fresh soybean hulls and (3) stored soybean hulls heated to 37 degreesC (E1), 55 degreesC (E2) and 80 degreesC (E3) or kept at room temperature (E4) for 16 h. Individual serum from 68 soybean asthmatic (SA) subjects, 30 nonallergic subjects and two serum pools made from 4 SA sera and 4 sera from asthmatics not sensitive to soybean were studied. All sera and serum pools were assayed for content of specific IgE (radioallergosorbent test) and IgG4 (ELISA). The following additional studies were done for extracts E1-E4: (1) SDS-PAGE, (2) SDS-PAGE/Western blot for specific IgE and IgG4 using both serum pools, and (3) study of the effects of heat on inhibiting activity of the extracts prepared from stored soybean hulls using the pool of SA sera. RESULTS: Test results demonstrated a reduced binding of specific IgE and IgG4 to fresh soybean hull extract compared to stored soybean hull extract, and an increased binding for heated extracts (E1-E3) compared to unheated ones (E4). Moreover, there was an increase in potency for IgE and IgG4 bindings for the heated (E1-E3) compared to unheated (E4) extract, as measured by the amount of protein to produce 50% inhibition. Several protein bands with a molecular weight (MW) higher than 20 kD were absent from the SDS-PAGE for E3 but were present in E1, E2 and E4, and a new protein band (MW 15.3 kD) appeared for E3 only. Two new protein bands, with MWs of 15.3 and 10 kD, which bind specific IgE, were present on Western blot and one of the 3 main soybean hull allergens, probably Gly m 2, disappeared in E3. IgG4 Western blot showed similar results, but only the 10 kD protein band was present. CONCLUSION: The results demonstrate that soybean hull allergenicity is affected by heat, and suggest that the heat generated during storage and transport of soybeans could generate 2 new allergen determinants or increases in epitope exposure as a result of conformational changes. The significance of these new IgE and IgG4 binding proteins has yet to be determined.

Purification and characterization of a soybean hull allergen responsible for the Barcelona asthma outbreaks. II. Purification and sequencing of the Gly m 2 allergen.

BACKGROUND: A low MW allergen from soybean hull, Gly m 1, with two isoallergens, Gly m 1 A and Gly m 1 B, was associated with the asthma outbreaks that occurred in Cartagena, Spain. Using sera of asthmatic epidemic patients (AEP) from Barcelona, three main soybean hull allergens, two of them with MWs and pIs identical to those reported for Gly m 1 A and Gly m 1 B, were identified. OBJECTIVE: The purpose of this study was to purify and to study the N-terminal amino acid sequence of the third allergen, which has a MW of 8 kDa. METHOD: The purification procedure combined the double dialysis method and preparative isoelectofocusing (IEF). Specific IgE determination to the fractions obtained demonstrated three peaks, one of them corresponding to the 8 kDa allergen. The pooled fractions containing this allergen were studied by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE), SDS-PAGE/Western blot and IEF/Western blot. Only a band with a MW of 8 kDa and a pI of 6 was obtained. Its allergenic activity was measured and it was demonstrated that the allergenicity of soybean hull correlates with the presence of the 8 kDa allergen. The N-terminal amino acid sequence of the first 20 amino acids, which was registered at the PIR Data Submission as the N-terminal partial sequence of Gly m 2, was determined according the Edman degradation method. RESULTS: Gly m 2 N-terminal amino acid sequence lacks homology with that reported for the allergen Gly m 1 but has a homology of 71% with a storage protein from cotyledon of Vigna radiata (cow pea) and 64% with a "disease response protein' from Pisum sativum (green pea). These results suggest that Gly m 2 in soybeans could protect against diseases which affect soybean plants. CONCLUSION: This study demonstrates the existence of another soybean hull allergen, Gly m 2, partially responsible for the soybean asthma outbreaks that occurred in Barcelona, Spain.

Specific immunoglobulins to soybean hull allergens in soybean asthma.

Soybean asthma, which occurred as an epidemic among patients in Barcelona, Spain, is associated with specific IgE to soybean hull allergens. The purpose of this study was to investigate the possible role of specific IgG, IgG subclasses, IgA, and IgM in the pathogenesis of soybean asthma. We studied 3 groups of subjects from Barcelona: group 1, 12 asthmatic epidemic patients; group 2, 23 asthmatic nonepidemic patients; and group 3, 32 nonallergic subjects. Specific IgE was determined by radioimmunoassay and specific IgG, IgG subclasses (1, 2, 3, and 4), IgA, and IgM by amplified enzyme-linked immunosorbent assay. Cross-inhibition studies were performed for specific IgE and IgG4. We partially characterized the soybean hull allergens that bind specific IgE, IgG, and IgG4 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis/Western blot. Percentage of positive results for the assays of the 8 Igs are as follows: for group 1, 100% (IgE), 75% (IgG), 16.6% (IgG1), 8.3% (IgG2), 0% (IgG3), 66.6% (IgG4), 25% (IgA), and 25% (IgM); for group 2, 4.3% were positive for specific IgE only; and for group 3, 0% (IgE), 0% (IgG), 6.2% (IgG1), 9.4% (IgG2), 9.4% (IgG3), 9.4% (IgG4), 6.2% (IgA), and 6.2% (IgM). The correlation between the specific IgE and the other specific Igs was significant between IgE and IgG4 in group 1 only (r = 0.752, p < 0.01). Cross-inhibition studies demonstrated a higher inhibitory capacity for IgG4 than for IgE. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/ Western blot demonstrates three low molecular weight protein bands that bind specific IgE, IgG, and IgG4. This study suggests that specific IgG4 to soybean hull allergens plays a role in the pathogenesis of soybean asthma and corroborates the role of specific IgE in the same disease.