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A double-immunostaining halogen immunoassay was developed to identify aerosolized conidia, hyphae, and fragments of Alternaria alternata by using an anti-Alternaria polyclonal antiserum, while, simultaneously, allergy to these components was concurrently determined by using human immunoglobulin E antibodies.

Background: More than 100 genera of fungal conidia are currently recognized as sources of allergens. The contribution of other fungal genera plus airborne fungal hyphae and fragmented conidia to allergic diseases is poorly understood., Objective: We sought to investigate the expression of allergens from airborne wild-type fungi using the Halogen immunoassay, which uses allergic serum IgE to immunostain immobilized allergens extracted from individual fungal particles., Methods: Airborne fungi were collected onto mixed cellulose ester protein-binding membranes for 2.5 hours with volumetric air pumps. Collected fungi were incubated overnight in a humid chamber to promote the germination of conidia. The membranes were laminated with an adhesive cover slip and immunostained with an Alternaria species-sensitive serum IgE pool. The samples were examined by means of light microscopy, and positively immunostained fungal particles were classified and counted., Results: All air samples contained fungal hyphae that expressed soluble allergens and were significantly higher in concentration than counts of conidia of individual well-characterized allergenic genera (P < .05). Resultant immunostaining of fungal hyphae was heterogeneous, and approximately 25% of all hyphae expressed detectable allergen compared with nonstained hyphae (P < .05). Fungal conidia of 10 genera that were previously uncharacterized as allergen sources were shown to demonstrate IgE binding to expressed antigens and accounted for 8% of the total airborne conidia count., Conclusions: Our analysis of wild-type fungi collected indoors presents a new paradigm of natural fungal exposure, which, in addition to commonly recognized species, implicates airborne hyphae, fragmented conidia, and the conidia of a much more diverse range of genera as airborne allergens.

Airborne fungi are ubiquitous in the environment and human exposure is inevitable. Such fungi differ greatly in their taxonomic, physical, ecological, behavioral, and pathogenic characteristics. Many strategies have evolved to sample, identify and interpret fungal exposure and their choice is determined by the hypotheses involved. While fungi can be sampled directly from surfaces, results do not generally reflect human exposure. For this reason, airborne spores are commonly sampled, by either filtration or impaction, using volumetric air samplers. Identification is commonly performed by either culture on nutrient medium or light microscopy using morphological criteria, although new techniques using DNA probes or characteristic antigens or toxins continue to be developed. Interpretation of such exposure data is both complex and contentious, but while there are numerous recommendations there is no consensus on exposure thresholds. A better understanding of the complex pathogenic roles of fungi and susceptibilities of their hosts will enable refinement of techniques for sampling and interpretation.

Background: Simple, inexpensive methods of sampling from allergen reservoirs are necessary for large-scale studies or low-cost householder-operated allergen measurement., Methods: We tested two commercial devices: the Indoor Biotechnologies Mitest Dust Collector and the Drager Bio-Check Allergen Control; two devices of our own design: the Electrostatic Cloth Sampler (ECS) and the Press Tape Sampler (PTS); and a Vacuum Sampler as used in many allergen studies (our Reference Method). Devices were used to collect dust mite allergen samples from 16 domestic carpets. Results were examined for correlations between the sampling methods., Results: With mite allergen concentration expressed as microg/g, the Mitest, the ECS and the PTS correlated with the Reference Method but not with each other. When mite allergen concentration was expressed as microg/m2 the Mitest and the ECS correlated with the Reference Method but the PTS did not. In the high allergen conditions of this study, the Drager Bio-Check did not relate to any methods., Conclusions: The Mitest Dust Collector, the ECS and the PTS show performance consistent with the Reference Method. Many techniques can be used to collect dust mite allergen samples. More investigation is needed to prove any method as superior for estimating allergen exposure.

Background: Prototype nasal filters were developed to collect inhaled pollen. This study evaluated the efficacy of the filters for prevention of rhinitis symptoms during acute outdoor pollen exposure., Methods: A randomized double-blind design was used. Subjects (n=46) with a history of autumn exacerbation of rhinitis and positive skin test to ragweed, Bermuda and/or Bahia grass wore either active or placebo nasal filters for 2 h in autumn in a park containing these species. Major and Total Symptoms scores were recorded at 0, 30, 60, 90 and 120 min., Results: Subjects wearing active nasal filters had significantly reduced scores, at all time-points compared with placebo group (all P <0.05). Of 14 individual symptoms measured, seven were significantly reduced (number of sneezes, runny nose, itchy nose, sniffles, itchy throat; itchy eyes and watery eyes) and another three showed a trend towards lower severity. The nasal filters also enabled the resolution of existing symptoms. Maximal difference in symptoms was seen immediately after subjects had spent 20 min sitting beside a large patch of ragweed., Conclusion: This is the first clinical trial of a nasal filter. The results suggest it has potential for enhancing rhinitis management during acute allergen exposure.

Background: Two factors thought to influence the risk of asthma are the promoting effect of sensitization to house dust mites and the preventive effect of increased omega-3 fatty acids. Although house dust mite allergen avoidance has been used as a preventive strategy in several trials, the effect of omega-3 fatty acid supplementation in the primary prevention of asthma and allergic disease is not known., Objective: To measure the effects of dietary supplementation with omega-3 fatty acids and house dust mite allergen avoidance in children with a family history of asthma., Methods: A total of 616 children at high risk of asthma were enrolled antenatally in a randomized controlled trial, and 526 children remained in the trial at age 3 years. The outcomes were symptoms of allergic disease and allergen sensitization., Results: There was a significant 10.0% (95% CI, 3.7-16.4) reduction in the prevalence of cough in atopic children in the active diet group ( P=.003; number needed to treat, 10) but a negligible 1.1% (95% CI, -7.1 to 9.5) reduction cough among nonatopic children. There was a 7.2% (95% CI, 10.11-14.3) reduction in sensitization to house dust mite in the active allergen avoidance group ( P=.05; number needed to treat, 14). No significant differences in wheeze were found with either intervention., Conclusion: These results suggest that our interventions, designed to be used in simple public health campaigns, may have a role in preventing the development of allergic sensitization and airways disease in early childhood. This offers the prospect of reducing allergic disease in later life.

Background: Measurement of exposure to the dust mite allergen Der p 1 is important in asthma research and is potentially useful in managing asthma. As no single measure can capture all characteristics of an exposure, it is important to recognize differences in the available methods of measuring exposure to Der p 1., Methods: Fourteen bedrooms and living rooms were sampled using four methods for 1 week. Airborne allergen was sampled by static Institute of Occupational Medicine samplers. Settling dust was collected on Petri dishes and an adhesive-membrane system (A-book). Vacuumed reservoir dust samples were collected from floors at the end of 1 week. Der p 1 was measured in all samples by enzyme-linked immunosorbent assay, except A-books, in which it was measured by Halogen immunoassay., Results: All four methods intercorrelated moderately (r range = 0.40-0.64, P = 0.04), except between allergen in reservoir dust (as microg/m2 and microg/g dust) and settling dust by Petri dishes (P = 0.2). Reservoir allergen, expressed as microg/m2, did not correlate with any measure, except reservoir allergen expressed as microg/g (r = 0.39, P = 0.04). No differences in these associations occurred between bedrooms and living rooms., Conclusions: While the four methods examined correlated moderately, all have practical advantages and difficulties. No method can be considered as ideal for measuring individual exposure. For practicality, use of vacuum cleaner and Petri dish methods are recommended.

Background: In temperate climates, exposure to house dust mite (HDM) allergens is the strongest environmental risk factor for childhood asthma. Environmental modifications to limit exposure have the potential to reduce the prevalence of asthma. The aim of this study was to reduce allergen exposure for children at high risk of developing asthma., Methods: A total of 616 pregnant women were randomized to HDM intervention and control groups. The control group had no special recommendations whereas the intervention group was given allergen impermeable mattress covers and an acaricidal washing detergent for bedding. Children were visited regularly until 18 months of age to have dust collected from their bed., Results: Der p 1 concentrations in the control group increased from 5.20 microg/g at 1 month to 22.18 microg/g at 18 months but remained low in the intervention group, ranging from 3.27 microg/g at 1 month to 6.12 microg/g at 18 months., Conclusions: In a high HDM allergen environment, a combined approach using physical barriers and an acaricidal wash, is effective in reducing HDM allergen concentrations in bedding. However, even with these control measures in place, HDM allergen levels remained high by international standards.

Background: Allergens dispersed by airborne fungal spores play an important but poorly understood role in the underlying cause and exacerbation of asthma. Previous studies suggest that spores of Alternaria and Aspergillus release greater quantities of allergen after germination than before germination. It is unknown whether this is true of other allergenic fungi., Objective: Our purpose was to investigate the release of allergen from a range of individual fungal spores before and after germination., Methods: Allergen expression from spores of Alternaria alternata, Cladosporium herbarum, Aspergillus fumigatus, Botrytis cinerea, Epicoccum nigrum, Exserohilum rostratum, Penicillium chrysogenum, Stemphylium botryosum, Curvularia lunata, Trichoderma viride, and Bipolaris spicifera was examined by halogen immunoassays through the use of pooled serum IgE from patients allergic to fungus. Spores were deposited onto protein-binding membranes direct from culture. To germinate spores, samples were incubated in high humidity at room temperature for 48 hours. Ungerminated and germinated samples were then laminated with an adhesive film and immunostained by the halogen assay. The samples were examined by light microscopy, and positive counts (haloed particles) were expressed as percentages of total spores., Results: For 9 of 11 species, between 5.7% and 92% of spores released allergen before germination. Spores of Penicillium and Trichoderma did not release detectable allergen. After germination, all spores that germinated had allergen elution from their hyphae. Eight of 11 species showed a significant increase (P <.05) in the percentage of spores eluting detectable allergen. Localization of allergen along the hyphae varied with species, such that some eluted allergen mainly from hyphal tips and septal junctions whereas others eluted allergen along the entire length., Conclusions: Increased elution of allergen after germination might be a common feature of many species of allergenic fungi. Although allergens from both spores and hyphae were recognized by human IgE, the extent to which human exposure occurs to allergens eluted from inhaled spores or from hyphae that germinate after deposition in the respiratory tract remains to be explored. The patterns of allergen expression might affect the clinical response to such exposure.

Background: Observational studies have linked house dust mite (HDM) exposure and dietary fatty acid intake with asthma in childhood. However, definitive evidence of their role in the etiology of asthma requires a randomized controlled trial., Objective: We hypothesized that the incidence of asthma and allergy in high-risk children would be reduced by avoidance of HDM allergens, supplementation with omega-3 fatty acids, or the combination of these strategies. We present the results of an interim analysis reporting outcomes assessed at 18 months., Methods: A total of 616 pregnant women were randomized to an HDM avoidance intervention, comprising the use of impermeable mattress covers and an acaricide or control and the use of an oil supplement, margarines, and cooking oils containing high levels of omega-3 fatty acids or control. Atopic status was measured by skin prick testing. Symptoms, diagnoses, and medication histories were elicited by means of parental interviews., Results: The diet intervention resulted in a 9.8% absolute reduction (95% CI, 1.5-18.1; P =.02) in the prevalence of any wheeze and a 7.8% absolute reduction (95% CI, 0.5-15.1, P =.04) in prevalence of wheeze of >1 week, but it had no effect on serum IgE, atopy, or doctors' diagnosis of asthma. The HDM avoidance intervention did not affect these outcomes but was associated with a lower use of oral steroids., Conclusion: Increasing dietary omega-3 fatty acids might have a beneficial effect on the prevalence of wheeze during the first 18 months of life. Follow-up to age 5 years, when the effect of the interventions on asthma risk will be assessed, is underway.

Background: The fungus Alternaria alternata contains potent allergens, and sensitization to these allergens is associated with a high risk of respiratory disease. The influence of genetic regulation on sensitization to Alternaria is unknown., Objective: To determine the influence of genetic factors on IgE responses to specific allergens of Alternaria., Methods: The concordance of skin prick test (SPT), radioallergosorbent test (RAST) and IgE-binding profiles of sera were examined from a large cohort of monozygotic and dizygotic twins., Results: Casewise concordance for a positive SPT response was monozygous (MZ) 66%: dizygous (DZ) 40% (P = 0.002). Logistic regression confirmed that casewise concordance was significantly stronger between MZ than DZ pairs. Immunoblotting against an Alternaria extract revealed 19 allergenic bands. The differences in concordance between the different bands were not significant for either the MZ (P = 0.97) or DZ (P = 0.84) groups. The pooled MZ : DZ difference in concordance was just significant (P = 0.049), suggesting an overall genetic effect on the response to Alternaria. This was reinforced by the comparison of the MZ and DZ correlations for total number of bands recognized (MZ r = 0.65; DZ r = 0.37, P = 0.015). Overall, there was a moderate correlation between the individual SPT weal size and RAST score (r(2) = 0.41) and a substantial correlation between the number of immunoblotted bands and RAST scores (r(2) = 0.79)., Conclusion: There is a strong genetic influence on IgE response to the mixture of Alternaria allergens and a lesser effect on IgE response to individual allergens.

Background: Although allergy to latex is a well-characterized phenomenon, some hospitals continue to provide staff with powdered latex gloves as an option to low- or non-powdered gloves., Objective: We aimed to measure the extent to which inhalation of latex particles could be reduced by the use of protective masks or by replacing powdered latex gloves with non-powdered latex gloves., Methods: Twenty healthcare workers in a hospital setting wore nasal air samplers (NAS) and Institute of Occupational Medicine (IOM) samplers for four 20-min periods. Subjects wore powdered gloves, non-powdered gloves and no gloves during three sampling periods, and in the fourth, subjects applied an aerosol barrier face-mask or a particulate face-mask (N95) while wearing powdered gloves. All samples were stained for particles bearing Hev b 5 allergen by the Halogen assay., Results: All subjects inhaled Hev b 5 bearing particles in all sampling periods. IOM samplers collected particles at 70% of the rate of NAS. The number of particles inhaled while wearing powdered gloves was 23.8-fold higher than when not wearing gloves and 9.7-fold higher than when wearing non-powdered latex gloves (P < 0.0001). Wearing an aerosol barrier mask did not significantly reduce the number of particles inhaled (P = 0.108), while use of particulate masks significantly reduced the number of particles inhaled by 17.4-fold (P = 0.003)., Conclusions: Use of non-powdered gloves is the most effective method of reducing occupational aeroallergen exposure to latex arising from gloves. However, secondary protection using particulate masks is a valid alternative, and may be helpful for preventing respiratory sensitization.

Background: Assessment of personal exposure to dust mite allergen has relied on proxy measures. Only recently has a means to directly measure inhaled allergen particle number become available (the intra-nasal air sampler)., Objective: To quantify inspired dust mite group 1 and group 2 allergen-bearing particles in bed in undisturbed conditions prior to sleep by nasal air sampling and to investigate the relationship between inhaled particles and reservoir allergen levels., Methods: Twelve volunteers wore nasal samplers in bed for 6 evenings, nose-breathing in undisturbed conditions. Allergen-bearing particles ('halos') were detected by immunostaining for Der p 1, Der p 2, or Der p 1 and Der p 2 together, and counted by light microscopy. Count data were square root transformed for analysis of variance. Mattress dust samples were assayed for Der p 1 and Der p 2 concentrations., Results: Square root detransformed mean particle counts per 30-min sample were: Der p 1, 4.22; Der p 2, 5.9; Der p 1 + Der p 2, 4.87; and for all samples, 5.01, with no difference between the groups. With replicate samples, halo number correlated significantly with mattress allergen concentrations (Der p 1 r = 0.80, P < 0.01; Der p 2 r = 0.68, P < 0.02)., Conclusion: Nasal air sampling can be used to quantify nocturnal Der p exposure in undisturbed conditions in an area with moderate exposure to mite allergen and can provide a direct measure of inhaled mite allergen. The choice of either Der p 1 or Der p 2 is appropriate for this purpose.

Background: IgE-mediated responses to natural rubber latex allergens have become a major health problem among recognized risk groups., Objective: The purpose of this investigation was to measure the amounts of Hevea brasiliensis latex allergen (Hev b 1) inhaled and deposited on surfaces when latex or vinyl gloves were worn and compare the results with the conventional measures (breathing zone samplers) of occupational exposure., Methods: Hev b 1 exposure was measured by nasal sampling and breathing zone sampling. Latex allergen exposure was generated by having each subject don a pair of powdered latex examination gloves and continuing his or her normal daily activity for 30 minutes. By means of adhesive tape, surface dust samples were collected from the surfaces of gloves, the subject's hands, and work areas. Sampling was performed with subjects wearing no gloves, subjects wearing powdered vinyl gloves, subjects wearing powdered latex gloves, and nearby colleagues wearing latex gloves. All samples were assayed through use of the HALOgen assay (Inhalix, Sydney, Australia) with a Hev b 1-specific mAb. Particles transporting latex allergen were identified by a surrounding immunostain halo, and these were quantified and reported as total numbers of particles inhaled, airborne, or found on surface areas evaluated., Results: Study subjects inhaled 26 times more allergen when powdered latex gloves were worn than under the "no glove" and powdered vinyl glove conditions. During the same period, Hev b 1 particle levels measured in the ambient air through use of the breathing zone sampler increased by 24-fold. The median numbers of particles carrying Hev b 1 allergen per square centimeter on the surface of the hands after the wearing of latex and vinyl gloves were 1964 and 5, respectively. Latex allergen was physically associated both with cornstarch granules and with larger dust particles having a darker, more irregular appearance., Conclusion: In a laboratory where gloves are worn for protection, the use of latex gloves resulted in a 26-fold increase in inhaled latex allergen over background levels measured while vinyl gloves were worn as controls. Low levels of latex exposure also occurred when vinyl gloves or no gloves were worn; the reasons for this are under investigation.

Allergen concentration in dust collected from a single square meter of carpet has been widely used as a proxy for aeroallergen exposure in a room. However, little is known about the horizontal distribution of allergens in carpeting or whether sampling several smaller sites might better approximate the average level. Eight bedrooms and 4 lounge rooms were divided into squares of 0.25 m(2), and the vacuumed dust was assayed for mite allergen (Der p 1). The proximity of each square to windows, doors, and beds was determined. In each room, the total allergen for each of 3 randomly chosen sets of 4 adjacent squares (forming a 1-m(2) area) was compared with that for each of 3 randomly chosen sets of 4 nonadjacent squares. The reliability of measuring allergen concentration from a single 0.25-m(2) area was modest (intraclass correlation coefficient [ICC] = 0.52), indicating wide-ranging allergen concentrations across short distances within each room. The reliability of the measurements based on 4 nonadjacent squares (ICC = 0.782) was higher than that of the measurements based on 4 adjacent squares (ICC = 0.67). Der p 1 concentrations decreased with distance from doors (paired partial correlation coefficient, -0.1217; P = 0.03). We recommend sampling 4 nonadjacent 0.25-m(2) areas that are not equidistant from the door when reservoir concentrations of Der p 1 are being measured.

Background: There is limited information about the removal of allergens by laundry washing., Objective: The purpose of this investigation was to determine the dynamics of the removal of mite allergen (Der p 1) and cat allergen (Fel d 1) from bed dust during simulated laundry processes., Methods: Three studies were performed. The first compared combinations of 4 laundry agents (water alone, soap, detergent with enzymes, and detergent without enzymes), 4 temperatures (15 degrees, 25 degrees, 45 degrees, and 60 degrees C), and 3 extraction times (5, 20, and 60 minutes). The second study examined allergen extraction by 11 common brands of detergents at 25 degrees and 45 degrees C for 5 minutes. The third study compared 4 detergents containing enzymes before and after the denaturation of their enzymes. To measure the quantity of allergens extracted, each study used an ELISA assay as well as a more sensitive but semiquantitative Halogen immunoassay to detect any allergens remaining after the simulated laundry extraction., Results: Study 1 showed that detergents extracted more of both Fel d 1 and Der p 1 than either soap or water alone and that almost all allergens were extracted within 5 minutes at 25 degrees. However, washing at 60 degrees C extracted slightly more Fel d 1 and denatured Der p 1, resulting in lower residual amounts of both allergens. Study 2 showed that all of the commercial detergents performed similarly. Study 3 showed that the presence of enzymes in detergent formulations did not produce a significant effect on the extraction of allergens., Conclusion: Using detergent solutions at 25 degrees for at least 5 minutes was sufficient to extract most mite and cat allergen from dust of bedding.

The Childhood Asthma Prevention Study is a randomized controlled trial to measure whether the incidence of atopy and asthma can be reduced by house dust mite allergen reduction, a diet supplemented with omega-3 fatty acids, or a combination of both interventions. Six hundred and sixteen pregnant women whose unborn children were at high risk of developing asthma because of a family history were randomized prenatally. Study groups are as follows: Group A (placebo diet intervention, no house dust mite reduction), Group B (placebo diet intervention, active house dust mite reduction), Group C (active diet intervention, no house dust mite reduction), and Group D (active diet intervention, active house dust mite reduction). The house dust mite reduction intervention comprises use of physical and chemical methods to reduce allergen contact. The dietary intervention comprises use of a daily oil supplement from 6 months or at onset of bottle-feeding, and use of margarine and cooking oils based on sunflower or canola oils to increase omega-3 dietary intake. Data is collected quarterly until the infant is 1 year old and then half yearly until age 5 years. Questionnaires are used to collect respiratory illness history and information about diet and home environment. Dust is collected from the child's bed and bedroom and playroom floors. Blinded assessments are conducted at 18 months, 3 years, and 5 years. Skin prick tests to common allergens, blood tests, and detailed illness, medication use, and vaccination histories are collected. Primary outcomes will be the development of allergic sensitization and the presence and severity of asthma. This study is designed to measure the effectiveness of allergen reduction and dietary supplementation, both separately and in combination, for the primary prevention of atopy and asthma. The results of this study may have important implications for public health policies to reduce the incidence of childhood asthma. Control Clin Trials 2001;22:333-354

Allergen released from individual spores of the fungus Alternaria has not been investigated. Germination of spores has been suggested to increase allergen release. This study examined allergen released from individual spores and the effect of germination on allergen availability. Allergen release was determined with the Halogen (Inhalix, Sydney, Australia) immunoassay, by use of serum IgE from Alternaria -sensitized subjects and 3 Alt a 1-specific antibodies. Not all spores released allergen. Germination of the spores significantly increased the proportion that released allergen (P < .0001 for all antibodies). Alt a 1 may be a minor contributor to the total allergen released from spores except when spores have germinated. How these results reflect the allergen content of spores in the air that we breathe requires investigation.

Background: In inland NSW, Australia, allergic sensitization to the fungi Alternaria and Cladosporium and to pollen is common and an important risk factor for asthma., Objective: We report the results of a series of experiments designed to assess the nature of personal exposure to these airborne allergenic particles. We have tested the effect of exposure conditions and level of activity on measurements of the personal exposure., Method: Personal Air Samplers (PAS) and Nasal Air Samplers (NAS) were employed. NAS are fitted just inside the nose and collect inhaled particles by impaction, while the PAS use a pump-operated filter with constant air flow (2 L/min). Thirty-three subjects (adults and children) used both NAS and PAS simultaneously for four one hour periods during which they performed activities or rested, both inside and outside their homes. Samples were analysed by light microscopy. Alternaria spores, Cladosporium spores, grass pollen and nongrass pollen were counted., Results: Both samplers detected substantial variation in exposure between subjects. Between members of the same household, the intrahouse correlation coefficient ranged from < 0 - 0.38. Levels of pollen grains and fungal spores inhaled were higher during periods of activity than during rest, and higher while subjects were outdoors than indoors. During the active outdoor period, the number of Alternaria spores inhaled ranged from 4 to 794 (median 11) spores/hr, Cladosporium from 0 to 396 (median 4) spores/hr, grass pollen from 0 to 81 (median 1) grains/hr and nongrass pollen from 0 to 72 (median 5) grains/hr., Conclusion: This is the first study to quantify individual inhaled levels of allergenic fungal spores and pollen under normal domestic circumstances. Exposure can be substantial and highly variable between individuals. The amount of particles inhaled relates both to location of the individual and activity being performed, independent of age group.

Background: Clothing has been proposed as an additional source of exposure to mite and cat allergens. Dispersal of allergen into public places has also been attributed to clothing., Objectives: We sought to study the contribution of various types of clothing on mite and cat exposure in a domestic environment. Also, we studied the ability of clothing to transfer allergen in a workplace., Methods: Personal exposure to mite and cat allergen from a range of clothing was measured by using intranasal air samplers in 11 homes. Five categories of clothing were tested. Wearing no upper clothing was the sixth category tested to distinguish the contribution of clothing over ambient background exposure. An adhesive tape was used to sample allergen from the surface of clothing, and reservoir dust samples were also collected. The above techniques were also used in the workplace to examine the amount of cat allergen transferred from cat owners to non-cat owners., Results: The amount of mite and cat allergen inhaled differed among the clothing types worn and whether they had been washed recently. Wearing a woolen sweater increased personal allergen exposure to cat and mite allergen by a mean of 11 and 10 times, respectively. Clothing items that were less frequently washed carried more allergen whether assessed by vacuuming or sampled with adhesive tape. This corresponded to the amount of allergen inhaled. We also found that cat levels on non-cat owners' clothing increased significantly at the end of a working day, which lead to the increase in their personal allergen exposure to cat., Conclusions: These studies strongly support the emerging model that personal clothing is an important source of both mite and cat allergen exposure. This article also demonstrates the importance of clothing as a means of distributing cat allergen into cat-free environments.

Background: House dust mite allergen levels in humid coastal regions of Australia are high, particularly in beds. Because high allergen levels in beds are associated with more severe asthma, reduction of levels may be important for asthma control., Objective: We tested the effectiveness of an acaricidal treatment of bedding in combination with occlusive mattress and pillow encasings in reducing allergen levels in children's beds in a community setting., Methods: A total of 14 beds of children were selected for the active intervention. In each home the bed of a sibling of nearest age was selected as the control. Dust was vacuumed from beds by using a standard protocol, and Der p 1 levels were measured by using ELISA. Adjacent settling dust was collected by using opened Petri dishes. The intervention consisted of encasing mattresses and pillows in occlusive covers and washing all bedding with Acaril, an acaricidal additive. The acaricidal wash was repeated twice in 7 households at 2-month intervals. Control beds were not treated., Results: The mean Der p 1 concentration at baseline was 27.9 microg/g in the active beds and 18.1 microg/g in the control beds. At 4 days after intervention, Der p 1 decreased to 3.2 microg/g and 15.7 microg/g in active and control beds, respectively. The average difference (active minus control) over the first 8-week cycle was 78.5% (P <.0001), and the difference over 3 washing cycles was 125.1% (P <.05). The mean rate of settling Der p 1 adjacent to the actively treated beds decreased from 24.4 ng.m(-2).d(-1) at baseline to 10.0 ng.m(-2).d(-1) after intervention (P <.01)., Conclusion: A substantial reduction in Der p 1 levels in beds and in airborne dust in a humid region with naturally high house dust mite allergen levels can be achieved and sustained in a community setting with use of occlusive covers and a rigorous washing routine.

Background: Exposure to outdoor allergens has commonly been estimated by collecting airborne particles with a Hirst-type spore trap and then using morphologic criteria to identify the intact pollen grains and fungal spores that are recognized as allergen sources. Several antibody-based blotting or fixation methods have also been developed that enable the counting of amorphous airborne particles carrying allergen, but none of these methods allow the ready association of the released allergen with the morphologically identifiable particle of origin. A method has been developed that uses pressure-sensitive adhesive tape to sample the airborne particles and then allows the immunoidentification of the specific particles that are the allergen sources., Objective: Our purpose was to visualize and immunostain the particles carrying pollen allergen that are collected with a volumetric spore trap., Methods: A Burkard sampler was used to collect airborne particles onto pressure-sensitive adhesive tapes. The particles were permanently fixed between the tape and a protein-binding membrane when the tape was laminated with the membrane. Allergens that elute from the particles onto the membrane were detected with a range of antibodies. Both the particle and associated immunostained allergen were viewed through the transparent tape for final microscopic identification., Results: Polyclonal and monoclonal antibodies and IgE from allergic patients stained allergens in the periphery of particles collected on the tapes. Individual pollen grains and paucimicronic particles were seen with halos of immunostained allergen surrounding them. When IgE was used, the density of immunostaining in the halo surrounding Lolium perenne pollen grains was found to be proportional to the level of Lolium-specific IgE. The method is highly sensitive and can be used to detect different airborne particles that carry allergen. Both the particle and the immunostaining can be subjected to a range of simple measurement techniques., Conclusion: Individual particles carrying allergens and antigens were visualized. These particles included intact pollen grains, paucimicronic particles, and fungal spores.

Background: Cockroach allergen is recognized as a causal factor for asthma. However, airborne cockroach allergen has not been detected in undisturbed conditions, and therefore the behavior and properties of airborne cockroach allergen have been poorly characterized. A new aeroallergen sampling method and sensitive system of immunoassay have been used to examine cockroach allergen exposure., Objective: Our purpose was to measure and characterize airborne cockroach allergens during normal domestic exposure in the homes of Sydney, Australia., Methods: Air sampling with Institute of Occupational Medicine, Edinburgh (IOM) samplers was performed in the living rooms of 10 houses during low- and no-disturbance environments. In addition, inhaled particles were collected by each home occupant during low domestic exposure with use of intra-nasal samplers that impact particles onto an adhesive surface. The particles collected on the IOMs and the intra-nasal samplers were immunostained with Bla g 1 monoclonal antibodies. Particle size, morphologic characteristics, and the relative Bla g 1 content of particles were estimated. Reservoir dust samples from the kitchen, living room, and bedroom were assayed by an ELISA. Two forms of repeatability of IOM air sampling were examined. The first measure tested the repeatability of 2 IOM samples collected simultaneously in the same room during low- and no-disturbance activities. The second measure examined the repeatability of IOM sampling over time on 10 consecutive days., Results: Bla g 1 was detected in reservoir dust samples taken from all homes (geometric mean 1.5 U/g, range 0.2-9.4 U/g). Inhaled particles containing Bla g 1 were detected during 1 hour of intra-nasal sampling in 8 of 10 homes during low disturbance. Cockroach particles were detected on all of the IOM samples collected for both 4-hour low-disturbance and overnight no-disturbance sampling environments. Particles containing Bla g 1 collected with the IOM samplers during low disturbance ranged in size from 3 to 350 microm. These particles are amorphous and irregular in shape, and a majority of the large particles were described as flakes (flat, transparent particles) and fibers (threadlike). A relationship was demonstrated between the allergen content of cockroach particles and their particle size. The larger particles elute more Bla g 1. The coefficient of repeatability for measurements made during low and no disturbance was 3.62 and 2.09, respectively. For measurements repeated over time at the same site, the coefficient of repeatability was 2.61. This represents the fold range within which 95% of pairs of measurements made at an interval of 1 day would be expected to lie., Conclusions: Airborne cockroach allergen is present in both undisturbed and low-disturbance environments in homes with relatively low reservoir levels of Bla g 1. In agreement with previous reports, airborne particles containing cockroach allergen (Bla g 1) are mainly associated with particles >10 microm. These particles are amorphous and irregular in shape and can be described as flakes and fibers.

Background: Measurement of personal exposure to Der p 1 aeroallergen has previously been limited by the low quantity of material collected by sampling systems and the assay sensitivity. This has meant that exposure could only be detected if long sampling periods were used or reservoir dust was artificially disturbed. We have developed a sampling method to sample true personal exposure and combined it with a novel method which is sensitive enough to measure allergen exposure over shorter time frames., Objective: To describe normal domestic exposure to dust mite allergen during a range of activities in houses in Sydney, Australia., Methods: Inhaled particles containing mite allergen Der p 1 were collected using a nasal air sampler which impacts particles (> approximately 5 microm) onto a protein-binding membrane coated with a thin, porous, adhesive film. The allergen is bound to the membrane in the immediate vicinity of the particle and detected by immunostaining with monoclonal antibodies specific for Der p 1. In addition, samples were collected using a standard Institute of Occupational Medicine (IOM) personal air sampler and the amount of eluted Der p 1 was assayed by ELISA., Results: The median number (range) of inhaled particles containing Der p 1 collected in each 10-min sampling period was: dust raising 5 (2-10); lying in bed, 0 (0-2); sitting on the bed, 1 (0-2); walking around the bedroom, 0 (0-2). This represented 0-5.1% of all particles captured. The Der p 1 concentration of floor and bed dust was 19.4 and 55.1 microg/g, respectively. The standard IOM personal sampler and ELISA were unable to detect Der p 1 for any of the activities performed., Conclusions: We were able to count individual allergen-carrying particles inhaled over short time periods, during different domestic exposure situations. This will offer new insight into several aspects of personal allergen exposure.

Background and Objectives: The ability to measure personal exposure to airborne grass pollen is important in the understanding of allergic diseases. Visual identification is time consuming and it is difficult to distinguish between many grass pollens morphologically. Although grass pollens share common allergenic determinants, we attempted to produce monoclonal antibodies that would distinguish between species, tribes and subfamilies of grasses which would allow immunodetection of pollens., Methods: Monoclonal antibodies raised against Lolium perenne were screened for specificity against an extended panel of grass pollen extracts using standard ELISA techniques and a novel particle blotting assay using whole pollen grains., Results: Antibodies showing specificity ranging from subfamily to part-tribe specificity were raised. The most specific monoclonal antibodies (numbers 4, 13 and 17) had reactivity to Lolium perenne and Festuca elatior but displayed little cross-reactivity to Phalaris arundinaceae and the rest of the Poeae tribe when tested by ELISA and no detectable cross-reaction when tested with particle blotting., Conclusion: Monoclonal antibodies that are functionally specific to only two grasses can be produced and used to discriminate between related grass species.

House dust mite (HDM) allergens are important factors in the increasing prevalence of asthma. The lung epithelium forms a barrier that allergens must cross before they can cause sensitization. However, the mechanisms involved are unknown. Here we show that the cysteine proteinase allergen Der p 1 from fecal pellets of the HDM Dermatophagoides pteronyssinus causes disruption of intercellular tight junctions (TJs), which are the principal components of the epithelial paracellular permeability barrier. In confluent airway epithelial cells, Der p 1 led to cleavage of the TJ adhesion protein occludin. Cleavage was attenuated by antipain, but not by inhibitors of serine, aspartic, or matrix metalloproteinases. Putative Der p 1 cleavage sites were found in peptides from an extracellular domain of occludin and in the TJ adhesion protein claudin-1. TJ breakdown nonspecifically increased epithelial permeability, allowing Der p 1 to cross the epithelial barrier. Thus, transepithelial movement of Der p 1 to dendritic antigen-presenting cells via the paracellular pathway may be promoted by the allergen's own proteolytic activity. These results suggest that opening of TJs by environmental proteinases may be the initial step in the development of asthma to a variety of allergens.

Background: Infancy may be a critical time for exposure to house dust mite allergens, when exposure to high levels can increase the risk of allergic sensitization and the development of asthma in later life., Objective: To measure house dust mite allergen (Der p 1) concentration in the infants' environment and examine lifestyle factors which may influence mite allergen exposure., Methods: Infants aged between 4 and 12 months (n = 134) from the western region of Sydney, Australia. participated. Reservoir dust samples were collected from four sites within each home: infant's bed, second bed (adult or second child's bed), lounge floor and sheepskins (where available). Settling aeroallergen was collected for 10-14 d in Petri dishes in the infant's room. Der p 1 was measured by ELISA. A questionnaire on types of bedding, sleeping and playing patterns of the infant was completed by the parents at the time of dust collection., Results: All infants were exposed to at least one site with Der p 1 concentrations greater than 10 microg/g fine dust. The mean settling aeroallergen level in the infants' room was 24 ng De p l/m2 day and this was weakly related to bed allergen levels (r=0.21, P<0.05). Mattress type had a weak effect on Der p 1 levels as measured in the whole bed (P = 0.07), while bed cover and bed type had no effect (P>0.6). The mean product of time spent at a site and its allergen concentration was highest for beds in 69% of infants., Conclusion: The high level of allergen exposure in the environment of this group of infants places them at an increased risk of early sensitization and development of asthma. Any strategy to reduce asthma prevalence should address these high and avoidable levels.

Background: In coastal Australia, mean house-dust-mite allergen concentration is 20-40 times higher in homes than in public buildings. Allergen concentrations in university colleges, which share some characteristics of both homes and public buildings, are not known. The study aimed to compare bed mite-allergen concentration in colleges with local homes., Methods: Mattress dust was collected from three colleges (n = 60 in each) and local homes (n = 68) during summer. Der p 1 was measured by ELISA. Information was collected on the floor plan of the colleges, cleaning practices, age of building, and orientation of room., Results: Most college mattresses (94%) had Der p 1 concentrations less than the mean of homes in the same climate. The geometric means of Der p 1 in the mattresses of the colleges were as follows: A, 8.9 micrograms Der p 1/g fine dust (95% CI 6.9, 11.5); B, 1.9 (1.5, 2.3); and C, 1.5 (1.2, 2.0), compared to homes, 22.5 (17.6, 28.7). The percentages of college mattresses with less than 2 micrograms/g were 7%, 48%, and 58%, respectively, compared to 4% for homes. Higher Der p 1 concentrations were weakly associated with age of building in college A, and orientation in college B. Der p 1 concentrations were independent of floor level and age of mattress., Conclusions: These findings indicate that low allergen concentrations are achievable without extreme hygiene and cleaning measures in a climate which supports mite proliferation in homes.

Background: Although some studies have shown genetic control of specific IgE responses to purified grass allergens, studies with other allergens have not supported this. The extent of such control for house dust mite (HDM) (Dermatophagoides pteronyssinus) allergens is unclear., Objective: We sought to determine the extent to which genetic factors control the specificity of IgE responses to individual HDM allergens by comparing the immunoblot patterns of IgE binding of serum from monozygotic and dizygotic members of a large cohort of Australian twins., Methods: HDM proteins separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis were immunoblotted with sera from 317 twin pairs in which at least one twin had at least a weak HDM skin test response. Concordance levels for IgE binding to the individual HDM components were compared in the subset of 142 pairs of twins in which both twins were allergic to HDMs (skin prick test wheal diameter, > 3 mm)., Results: Over all 36 blotted bands, the mean case-wise concordance was 41% for monozygotic twins and 17% for dizygotic twins. Of the components detected, only those of molecular weights 23 kd and 16 kd were significantly different between the groups (p < 0.01). Differences observed between the monozygotic and dizygotic twins could be partly explained by overall IgE hyperresponsiveness., Conclusion: Evidence for genetic control of IgE responses to 36 IgE-binding HDM components from a large sample of twins showed significant differences in concordance for two components and nonsignificant differences for several others. In the monozygotic twins, concordance never exceeded 67% for any band, and most monozygotic individuals recognized components their co-twin did not. Genetic control of overall atopy in monozygotic twins is far stronger than that controlling specific sensitization to HDM allergens.

Background: Both genetic and environmental factors are thought to contribute to specific IgE responses, however, the relative contribution of each in the responses to individual ryegrass pollen allergens is largely unknown even though some responses to allergens have been linked to certain HLA complexes., Objective: Using a large group of monozygotic and dizygotic twins, this study was designed to investigate the IgE binding profiles of individual ryegrass pollen (Lolium perenne) components to assess the relative contribution of genetic and environmental factors in determining IgE responses to specific allergens., Methods: Ryegrass pollen proteins were separated by electrophoresis and immunoblotted with sera from 191 pairs of twins where at least one sibling had a SPT > 2 mm to perennial ryegrass. Concordance levels for individual ryegrass pollen components were compared between monozygotic and dizygotic twins in a subset group where both twins had SPT > 3 mm to perennial ryegrass., Results: Immunoblotting revealed 23 individual IgE-binding components from ryegrass pollen. Although there was a significantly greater proportion of monozygotic twins with SPT wheals greater than 3 mm when compared with the dizygotic twins, the mean case-wise concordance for the immunoblot components was similar for both groups of twins. The mean case-wise concordance when at least four pairs of sera were involved was 44% for the MZ twins (n=11 components) and 45% for the DZ twins (n=12 components). We found no significant differences in concordance levels for any of the 23 individual components including allergens previously associated with HLA., Conclusion: Evidence for genetic control of allergen-specific IgE responses in a large population sample of twins to individual ryegrass allergens is limited, indicating that the IgE responses to specific ryegrass pollen allergens are determined largely by environmental factors.

Ribosomal DNA internal transcribed spacer (ITS) sequences were investigated in the genera Alternaria, Aspergillus, Cladosporium and Penicillium, to identify regions suitable for the design of genus- and species-specific oligonucleotide probes. The ITS regions of all four genera were quite distinct. However, we discovered that the genera Aspergillus and Penicillium displayed high similarity in most of the ITS2 region. Whilst interspecies similarity within Alternaria and Cladosporium was high in both ITS regions 1 and 2, ITS1 similarity within Aspergillus and Penicillium was lower by comparison. We conclude that: (1) the ITS2 region is highly conserved between Aspergillus and Penicillium, and (2) the ITS1 region will allow for the development of specific probes to distinguish between Aspergillus and Penicillium.

Objective: To determine the prevalence and nature of asthma in four rural Australian Aboriginal communities., Design: Cross-sectional population study., Setting: Four Aboriginal communities in Queensland, the Northern Territory and South Australia, Australia., Subjects: Data were collected from 1252 subjects aged 5-84 years in August 1990 and August/September 1991., Main Outcome Measures: Respiratory symptoms, measured by interview-administered questionnaire; airway hyperresponsiveness (AHR), measured by histamine challenge; and allergy, measured by skin-pick tests. AHR was a PD20FEV1 of histamine of less than or equal to 3.9 mumol., Results: The prevalence of AHR in the four communities ranged from 2.2% to 7.5% and significantly increased with age (chi 2 trend test: P < 0.05). The prevalence of current asthma was 0.5% among 8-12 year old children and 3.3% among adults. The overall prevalence of atopy in the four communities ranged from 21% to 34%. Allergy to cats, house dust mites or cigarette smoking was a risk factor for AHR, and cat allergy was a risk factor for current asthma., Conclusions: The prevalence of asthma in rural Aboriginal adults is low in comparison with the prevalence among non-Aboriginal Australians, and asthma in Aboriginal children is almost non-existent. The low prevalence of asthma is possibly due to environmental factors that influence the acquisition of atopy and AHR.

Background: To interpret individual measurements of house dust mite (HDM) allergen and to design and analyse HDM studies it is necessary to quantify the variability which is inherent in the measurement of this exposure., Objective: To estimate the repeatability of one method of HDM allergen measurement., Methods: We analysed data from one or more HDM allergen measurements in 215 houses included in four previous studies conducted in Sydney (a high allergen environment) and Busselton, Western Australia (a moderate allergen environment). Samples were collected from the bed by vacuuming above and below the sheets and inside the pillow case and from the bedroom and living room floors by vacuuming a 1 m2 area for 1 min. Extracts from aliquots of fine dust from each sample were assayed for HDM allergen Der p I using a monoclonal antibody enzyme linked immunosorbent assay (ELISA). The values for HDM allergen were positively skewed and the suitability of a log transformation was established by the resulting normal distribution and stable within-site variance., Results: The range of single determination (within which the true value lies with 95% certainty) was 3.1-fold for samples from the bed and 3.5-fold for samples from the floor. The coefficient of repeatability (the ratio beyond which a change between two estimates is established with 95% certainty) was 4.9 for the bed and 5.8 for the floor., Conclusion: We estimate that, to detect a twofold difference or change in allergen levels, 35 houses per group will be required in cross-sectional studies and 30 houses per group in parallel-group, randomized controlled trials. We recommend that beds be sampled by collecting dust from the layer of bedding below the bottom sheet. A single site within the bedroom floor may be taken as representative of this site but this is not true for the living-room floor.

House dust mite (HDM) allergen exposure and its relation to HDM allergy and asthma was assessed in a case-control study conducted over three seasons in 74 Sydney schoolchildren, 33 of whom were allergic to HDM and 12 of whom had current asthma. In each season histamine inhalation tests and skin prick tests were performed, symptom questionnaires were administered, and dust samples were collected. The mean concentrations of HDM allergen (in micrograms of Der p 1 per gram of fine dust) were: bed, 38.9 (95% confidence interval [CI], 31.8 to 47.5); bedroom floor, 22.4 (95% CI, 18.3 to 27.5); and lounge room floor, 13.7 (95% CI, 10.7 to 17.6). The mean of the highest allergen concentration in each house was 51.0 (95% CI, 43.2 to 60.1). All but two subjects had at least one site in all seasons with an HDM allergen concentration greater than 10 micrograms/gm, the proposed threshold for asthma symptoms. Subjects with allergy to HDM, symptoms of asthma, or airway hyperresponsiveness did not have higher HDM allergen concentrations in their house. In this study we were unable to test hypotheses concerning proposed thresholds for risk of sensitization and for risk of asthma symptoms because virtually all subjects were exposed to HDM allergen levels above the proposed thresholds.

To measure the association between changes in house dust mite (HDM) allergen Der p I exposure and changes in the severity of asthma, we re-analysed data from a clinical trial in which 34 HDM-allergic subjects with asthma (18 women, mean age 35 years) were followed for between 3 and 12 months. The concentration of Der p I in fine dust from the bed, the bedroom floor and the living room floor was measured at 3-monthly intervals along with assessment of subjects' spirometric function and airway hyperresponsiveness (AHR, measured by histamine inhalation test). Daily symptom scores, morning peak expiratory flow rate and peak flow variability were measured throughout the study period. The mean Der p I concentration in the bed at baseline was 25.4 micrograms/g (95% CI: 15.8-40.6). During the course of the study large within-subject fluctuations were observed in allergen concentrations and in the measurements of the severity of asthma. Changes in allergen concentration in the bed were significantly correlated with changes in AHR (P = 0.003) and symptom score (P = 0.04). Changes in allergen concentration in the living room floor were correlated with changes in symptom scores (P = 0.01). Although these correlations were significant the magnitude of the effect was relatively modest. We conclude that a large reduction in HDM allergen concentration, particularly in the bed, results in a modest reduction in AHR and improvement in symptoms in HDM-allergic subjects with asthma.

To test the effectiveness of a house dust mite (HDM) allergen avoidance strategy we conducted a randomized controlled trial in 35 atopic subjects with asthma, aged 13 to 60 living in Sydney - a high HDM allergen environment. After a 3 month run-in period, subjects were randomized to active allergen avoidance treatment (n = 17) or placebo (n = 18) groups and followed for 6 months. The active treatment involved placing impermeable covers over the mattress, pillows and duvet and spraying the remaining bedding, as well as the carpets and furniture, with a tannic acid/acaricidal spray. Subjects kept a daily record of symptoms and peak expiratory flow rates and had 3 monthly assessments of lung function and airway hyperresponsiveness (AHR). Dust samples were collected from the bed, the bedroom floor and the living room floor at 3 monthly intervals and 2 weeks after the treatment. Mean HDM allergen levels at baseline at these sites were, in the active group, 15.5, 9.6 and 10.2 micrograms Der p I/g of fine dust, and, in the placebo group 25.7, 11.8 and 6.3 micrograms/g. Two weeks after the allergen avoidance treatment the HDM allergen level in the beds was reduced to 29% of baseline (95% CI 16-50%, P = 0.038 compared with placebo), but was not significantly different at 3 or 6 months. There was also no significant effect of the allergen avoidance treatment on symptom scores, peak flow variability, lung function or AHR P > 0.1).(ABSTRACT TRUNCATED AT 250 WORDS)

Allergens produced by the house dust mites (family Pyroglyphidae) are probably the single most important allergens associated with asthma world wide. If exposure to these allergens in houses could be sufficiently reduced, then asthma symptoms may be markedly reduced and even prevented from being initiated. Only about half of the many attempts to reduce mite allergens in houses have shown any clinical benefit. One reason may be that exposure was not reduced enough--however exposure to mite allergens has never been measured in any trial. This review summarises previous allergen control trials and then provides an outline of allergen exposure, including the nature of exposure, the analytical methods available and the recognised risks of allergen exposure. This provides a perspective to evaluate the individual methods used to kill mites and to reduce exposure to the allergens. The object is to provide a framework to improve and develop allergen avoidance as an effective component of asthma management.

The beds, carpets and furnishings in 15 houses were sprayed with a solution containing tannic acid and an acaricide in an attempt to reduce allergen concentrations. Dust was collected from these sites for 4 weeks following spraying and the mite allergen Der p I concentration was measured and compared with baseline concentrations. In a subgroup of houses, counts of live mites and estimates of aeroallergen were also made. Four untreated houses were monitored over the same period. In dust samples collected 3 days after spraying, the mean concentrations of Der p I in beds, carpets and furniture were 23%, 28% and 26% of the pretreatment levels. All these reductions were significant compared to untreated controls. Samples collected 4 weeks after treatment were not significantly different to baseline for each group. After the initial reduction, the rate of increase in allergen concentration was significantly greater in the spray-only beds than in the beds which had been both sprayed and fitted with occlusive covers. Both aeroallergen and live mites continued to be detected in houses after treatment with the spray. These studies suggest that such sprays are only temporarily effective when applied at the manufacturer's recommended volumes and additional approaches are required to control the bulk of allergens in houses.

Cryostat sections of the house dust mite, Dermatophagoides pteronyssinus, were probed in fluorescent microscopy studies with rabbit polyclonal and mouse monoclonal antibodies specific for mite allergens including the major allergen, Der p I. Sections also were probed for allergens with sera from human mite-allergic subjects containing IgE antibodies to mite allergens and with lectins. Antibody binding was mainly to the gut lining and gut contents of the mite, although some specific labeling also was associated with the head region and cuticle. This is the first detailed localization of mite allergens in situ. The morphology of the mite was investigated using plastic embedded thin sections and was found to be similar to that previously described for D. farinae.

Analysis by protein blotting of sera from 96 different house dust mite-allergic subjects revealed previously unrecognized complexity of low molecular weight (MW) (less than 20 kD) IgE-binding proteins in extracts of whole bodies of Dermatophagoides pteronyssinus. Of 11 different IgE-binding components of MW less than 20 kD identified, two (MW approximately 16 kD and approximately 15 kD), showed both a high frequency (88% and 49% respectively) and a high intensity of IgE-binding. The approximately 16 kD component, identified as allergen Der p II, showed the highest frequency of IgE antibody reactivity of any of the major D. pteronyssinus allergens including Der p I and Der p III.