Personal exposure to house dust mite allergen in bed: nasal air sampling and reservoir allergen levels

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.     

House dust mite allergen exposure in infancy

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.     

Sequence polymorphisms of the Der p 3 house dust mite allergen

Background The trypsin-like protein Der p 3 is a major allergen of Dermatophagoides pteronyssinus. Like other vertebrate and invertebrate trypsin-like molecules, isoelectricfocusing studies with the natural Der p 3 protein have indicated that several isoforms exist. Objective To determine the extent of the sequence variation of the Der p 3 allergen and distinguish at the molecular level, whether the sequence isoforms represent allelic variants or multiple genes of the allergen. Methods Five cDNA clones of Der p 3 have been isolated from a λ gt 10 D. pteronyssinus library, using a radiolabelled polymerase chain reaction (PCR) Der p 3 P3WS1 probe and sequenced. Southern blot and inverse PCR analysis of Eco RI digested genomic DNA was performed. Results Southern blot analysis of Eco RI digested genomic DNA showed that the DNA encoding Der p 3 was located on a single 3.5 kb fragment and inverse polymerase chain reaction analysis (PCR) of this DNA showed that there was only a single Der p 3 gene on this 3.5 kb fragment. The nucleotide sequence of one of the clones was identical to the original Der p 3 P3WS1 clone and two clones ditfered only in their 3′untranslated sequences. The other two contained nucleotide changes which lead to several substitutions at the amino acid level, both conservative and non conservative. Clone 3 had 98.7% identity with Der p 3 P3WS1. One clone for which the full sequence was not available (clone 4) had only 84.4% identity with the original clone and is therefore consistent with an isoallergen. Conclusions These data along with our previous genomic sequence shows that for the most part, the Der p 3 allergen has only minor sequence variations (variants) although the isoallergen indicated by clone 4 needs further investigation. It is now evident that Der p 3 is encoded by a single gene and that most cDNA clones constructed from commercial mites show only minor sequence variation similar to that observed for the group I and group 2 house dust mite allergens.     

Influence of mattress characteristics on house dust mite allergen concentration

BACKGROUND: Exposure to a high level of house dust mite allergens (HDMAs) is considered as a risk factor for HDM sensitization and development of asthma in genetically disposed people. Mattresses are one of the most important sources of HDMA in people's living environment. OBJECTIVE: The aim of this study was to evaluate the association between mattress characteristics and HDMA concentrations on mattresses. METHODS: Dust samples of mattress surfaces were taken to evaluate the level of Der p 1 allergen. All participants filled in a questionnaire about the type of mattress, the type of covering (upper layer) of the mattress, dwelling characteristics and cleaning habits. Humidity and temperature of the bedroom were measured at the time of dust sampling. RESULTS: One hundred and sixty-eight questionnaires were filled in. Synthetic upper layer of the mattress was associated with a higher level of Der p 1 compared with cotton upper layer (2.6 vs. 0.8 microg/g Der p 1). Moreover, higher relative humidity (RH) was associated with significant higher concentrations and density of Der p 1. CONCLUSIONS: Two factors were associated with lower levels of Der p 1 found on mattresses, namely: a cotton upper layer of the mattress compared with a layer of synthetic material and lower RH at the time of sampling. As far as we know, the association between type of upper layer and concentration of Der p 1 has not been described before and could lead to the formulation of practical advices in order to reduce HDMA concentrations on mattresses.     

Nonlinear relations between house dust mite allergen levels and mite sensitization in farm and nonfarm children

BACKGROUND: Low sensitization rates to common allergens have been observed in farm children, which might be due to high exposure to microbial agents. It is not known how microbial agents modify the association between specific allergen exposure and sensitization. OBJECTIVE: To examine the relations between house dust mite allergen exposure and mite sensitization in farm and nonfarm children and to assess the effects of microbial agents levels on this association. METHODS: Major mite allergens of Dermatophagoides pteronyssinus (Der p 1) and Dermatophagoides farinae (Der f 1), endotoxin, beta(1,3)-glucans and fungal extracellular polysaccharides were measured in mattress dust of 402 children participating in a cross-sectional study in five European countries. Mite allergen (Der p 1 + Der f 1) levels were divided into tertiles with cut-offs 1.4 and 10.4 microg/g. Sensitization was assessed by measurement of allergen-specific immunoglobulin E against house dust mite. RESULTS: Prevalence ratios of mite sensitization for medium and high when compared with low mite allergen levels were 3.1 [1.7-5.7] and 1.4 [0.7-2.8] respectively. Highest mite sensitization rates at intermediate exposure levels were consistently observed across country (except for Sweden) and in both farm and nonfarm children. The shape of the dose-response curve was similar for above and below median mattress microbial agent levels, but the 'sensitization peak' appeared to be lower for above median levels. CONCLUSIONS: Our data suggest a bell-shaped dose-response relationship between mite allergen exposure and sensitization to mite allergens. In populations with high microbial agent levels and low sensitization rates, the curve is shifted down.     

Silverfish protein in house dust in relation to mite and total arthropod level

Two assays have been developed to measure arthropod levels in house dust. The first assay measures silverfish antigens. The second assay measures invertebrate tropo-myosin and gives a global assessment of the level of atthropod-derived material. These assays and a Der p 1 and Der p 2 assay were used to analyse 53 dust samples. In most dust samples the ratio of tropomyosin/Der p 2 was higher than in mite body extract, indicating that the assay measures other arthropods besides mites. Silverfish antigen was detectable in most of the dust samples. In many homes in which the inhabitants were unaware of the presence of silverfish, silverfish antigen was detectable. Therefore for information on exposure an immunochemical analysis is superior to a questionnaire.     

哮喘患儿家庭内尘螨过敏原分布特征及其临床意义

目的了解北京地区尘螨过敏性哮喘患儿家庭环境内尘螨过敏原含量分布特征,初步探讨尘螨过敏原暴露水平的临床意义。方法选取54例尘螨过敏性哮喘患儿,其中男性37例,女性17例;年龄3～16岁,平均年龄8岁2个月。采集患儿家庭中床垫、枕头、卧室地板、客厅地板及沙发的灰尘,采用酶联免疫吸附分析（ELISA）测定以上灰尘样本中户尘螨1组过敏原（Der p1）和粉尘螨1组过敏原（Der f1）的含量;应用荧光ELISA测定患儿血清尘螨特异性IgE浓度;评估患儿哮喘临床控制情况,应用化学发光法测定患儿呼出气一氧化氮浓度（FeNO）。结果采集灰尘样本255份,以中位数（最小值～最大值）表示尘螨过敏原含量,床垫、枕头和沙发灰尘样本中Derf1和Derp1的含量显著高于卧室地板和客厅地板灰尘样本中尘螨过敏原含量。Derf1平均含量为0.13μg/g,显著高于Derp1平均含量0.02μg/g（P〈0.05）。Derp1和Derf1联合暴露的最高含量平均为2.18（0.07～54.59）μg/g。Der p1和Der f1联合最高暴露含量≥10.00μg/g、2.00～10.00μg/g、0.05～2.00μg/g的例数分别为4例（7.4%）、24例（44.4%）、26例（48.1%）。其中未控制组患儿家庭内尘螨过敏原最高暴露水平为27.41（0.23～54.59）μg/g,均显著高于部分控制组和控制组哮喘患儿尘螨过敏原最高暴露水平1.66（0.07～26.27）μg/g、2.90（0.37～33.75）μg/g（P〈0.05）。不同sIgE浓度分级组间尘螨过敏原最高暴露水平的差异、不同FeNO浓度范围组间尘螨过敏原最高暴露水平差异均无统计学意义。结论北京地区尘螨过敏性哮喘患儿家庭尘螨以Der f1为主,床垫、枕头及沙发灰尘样本是Der p1和Der f1的主要来源;哮喘未控制者的尘螨过敏原最高暴露水平明显增高。     

Distribution of house dust mite allergen: comparing house dust mite allergen levels in dust samples collected from different sites on living room floors with smooth coverings

BACKGROUND: The distribution of house dust mite allergen (Der p1) in living rooms with smooth floor coverings, as measured in the middle compared with the border of the floor was investigated. It was hypothesized that activity causes displacement of Der p1, from the middle towards the border. METHODS: Dust samples from the middle and border of 50 floors with smooth coverings were collected and analysed on Der p1 content in a standardized way. RESULTS: The Der p1 exposure expressed as per unit area (ng/m2) showed that border samples contained significantly more Der p1 compared with middle samples (median: 2.57 vs 0.27, respectively, P = 0.023). Presence of pets and presence of more than two inhabitants increased the difference. When expressed as per unit weight of dust (ng/g), significant differences were only detected when comparing Der p1 content of samples collected in households with three or more inhabitants [median: 2 (border) vs 53 (middle), respectively; P = 0.035]. CONCLUSIONS: The Der p1 is unequally distributed on living room floors with smooth coverings, most likely because of displacement of dust from the middle towards the border due to activity. Expression as ng/g of dust and ng/m2 could not obviously be interchangeable.     

Effect of improved home ventilation on asthma control and house dust mite allergen levels

Background:  The warm, humid environment in modern homes favours the dust mite population, but the effect of improved home ventilation on asthma control has not been established. We tested the hypothesis that a domestic mechanical heat recovery ventilation system (MHRV), in addition to allergen avoidance measures, can improve asthma control by attenuating re-colonization rates.
Methods:  We conducted a randomized double-blind placebo-controlled parallel group trial of the installation of MHRV activated in half the homes of 120 adults with asthma, allergic to Dermatophagoides pteronyssinus . All homes had carpets steam cleaned and new bedding and mattress covers at baseline. The primary outcome was morning peak expiratory flow (PEF) at 12 months.
Results:  At 12 months, the primary end-point; change in mean morning PEF as compared with baseline, did not differ between the MHRV group and the control group (mean difference 13.5 l/min, 95% CI: −2.6 to 29.8, P  = 0.10). However, a secondary end-point; evening mean PEF, was significantly improved in the MHRV group (mean difference 24.5 l/min, 95% CI: 8.9–40.1, P  = 0.002). Indoor relative humidity was reduced in MHRV homes, but there was no difference between the groups in Der p 1 levels, compared with baseline.
Conclusions:  The addition of MHRV to house dust mite eradication strategies did not achieve a reduction in mite allergen levels, but did improve evening PEF.     

Molecular characterization of Der p 10: a diagnostic marker for broad sensitization in house dust mite allergy

Background Tropomyosins represent clinically relevant seafood allergens but the role of mite tropomyosin, Der p 10, in house dust mite (HDM) allergy has not been studied in detail. Objective To express and purify a recombinant Der p 10 with equivalent IgE reactivity as natural Der p 10 and to evaluate its IgE reactivity and allergenic activity in HDM‐allergic patients. Methods rDer p 10 was expressed in Escherichia coli, purified and characterized by mass spectrometry and circular dichroism. It was tested for IgE reactivity in 1322 HDM‐allergic patients. Detailed IgE‐reactivity profiles to six HDM allergens (Der p 1, 2, 5, 7, 10, 21) were established for subgroups of Der p 10‐positive and ‐negative patients. The allergenic activity of rDer p 10 was evaluated in basophil degranulation experiments. Results rDer p 10 is an α‐helical protein sharing IgE epitopes with nDer p 10. It is recognized by 15.2% of HDM‐allergic patients. Der p 10‐negative patients were primarily sensitized to Der p 1 and/or Der p 2, whereas Der p 10‐positive patients reacted to several other HDM allergens besides the major allergens (Der p 1, Der p 2) or showed a rather selective Der p 10 reactivity. The allergenic activity of Der p 10 was generally low but patients could be identified who suffered from clinically relevant HDM allergy due to Der p 10 sensitization. Conclusion and Clinical Relevance Der p 10 may be a diagnostic marker for HDM‐allergic patients with additional sensitization to allergens other than Der p 1 and Der p 2. Such patients may require attention when allergen‐specific immunotherapy is considered. Cite this as: Y. Resch, M. Weghofer, S. Seiberler, F. Horak, S. Scheiblhofer, B. Linhart, I. Swoboda, W. R. Thomas, J. Thalhamer, R. Valenta and S. Vrtala, Clinical & Experimental Allergy, 2011 (41) 1468–1477.     

Molecular cloning and immunological characterization of the house dust mite allergen Der f 7

Background The allergen Der p 7 from Dermatophagoides pteronyssinus has been defined by molecular cloning and shown to be an important specificity in 50% of miteallergic patients. Objective This study compares the cDNA sequence and serological reactivity of Der f 7 from D. farinae with Der p 7. Method cDNA encoding Der f 7 was amplified by polymerase chain reaction. sequenced and expressed as a fusion with glutathione-S-transferase for IgE and monoclonal antibody binding studies. Results Der f 7 cDNA encoded a 213 polypeptide containing a predicted 17 amino acid leader sequence, no cysteines and a single N-glycosylation site similar to Der p 7. The predicted 196 residue mature polypeptide had 86% identity lo Der p 7 and a calculated molecular weight of 22 34SDa. No homologues were found in searches of the data banks. The Der f 7 fusion protein showed a single band of 46 k Da by sodium dodccyl snlfute-polyaerylamide gel electrophoresis (SDS-PAGF) and reacted with IgE antibodies in 19/41 (46%) of sera from asthmatic children. The degree of binding was usually 30% of that to Der p 7 consistent with the exposure of the patients to D. Pteronyssinus. Monoclonal antibodies (WH9 and WH22) against Der p 7 reacted with Der f 7 but inhibition studies showed a 10-fold difference in reactivity. Conclusion Der f 7 has a predicted 213 residue polypeptide with 86% homology and serological cross reactivity to Der p 7.     

Cord blood mononuclear cell cytokine responses in relation to maternal house dust mite allergen exposure

BACKGROUND: Cord blood mononuclear cells have demonstrated specific immune responses to environmental allergens. OBJECTIVE: To establish whether the nature of this response is related to the level of maternal antenatal exposure to house dust mite (HDM) allergen and, hence, whether antenatal allergen avoidance may have a role in the prevention of allergic sensitization in children. METHODS: Children with a family history of asthma were recruited antenatally as subjects in a randomised controlled trial: the Childhood Asthma Prevention Study. HDM allergen (Der p 1) concentrations were measured in dust collected from the maternal bed at 36 weeks gestation. Cord blood mononuclear cells were stimulated in culture, separately, with phytohaemaglutinin (PHA) and HDM extract. Cytokine IL-4, IL-5, IL-10 and IFN-gamma concentrations in supernatant were measured by ELISA. mRNA signals for these cytokines were measured using RT-PCR. RESULTS: The median concentration of HDM allergen was 18.4 microg/g (interquartile range 7.3-35.3 microg/g). Median concentrations of IL-4, IL-5, IL-10 and IFN-gamma, after PHA stimulation were 4, 19, 401 and 1781 pg/mL, respectively. After HDM allergen stimulation the median concentrations were 0, 0, 20 and 14 pg/mL, respectively. The distribution of mRNA cytokine signals was similar. Neither cytokine protein concentrations nor cytokine mRNA signal levels were correlated with the concentration of HDM allergen in the mothers' beds at 36 weeks gestation. CONCLUSION: These findings do not support the view that the prevention of allergic disease in children requires the institution of HDM avoidance interventions during pregnancy.     

Predictors of high house dust mite allergen concentrations in residential homes in Sydney

BACKGROUND: In parts of coastal Australia, house dust mite allergen concentrations in homes are often very high with at least 80% of homes in Sydney exceeding concentrations of 10 microg of allergen per gram of fine dust. In this study, we report the relation between characteristics of the home environment and house dust mite allergen concentrations at three sites in Sydney homes. METHODS: A total of 616 families were recruited as part of the Childhood Asthma Prevention Study (CAPS). Information about the home environment and structural aspects of the home was collected using a questionnaire. Samples of dust were collected from the parents' bed, the bedroom floor and the living room floor and assayed for Der p 1. RESULTS: A total of 68% of participants' beds, 65% of bedroom floors and 56% of living room floors had Der p 1 concentrations above 10 microg/g, with the highest concentrations of allergen in the bed. The most significant predictor of high Der p 1 concentrations in the bed and floors was the age of the home. We also found that beds with mattresses over two years old and with woollen or synthetic blankets or synthetic quilts had higher Der p 1 concentrations. Carpeted floors had higher Der p 1 concentrations than hard floors. CONCLUSION: The finding that high Der p 1 allergen concentrations in homes with carpets and older mattresses indicates that control strategies directed at these sources are likely to be effective in reducing exposure. Alternatives such as the use of house dust mite impermeable mattress encasings on older mattresses may also be effective in reducing exposure.     

Genetic polymorphisms of major house dust mite allergens

BACKGROUND: Polymorphic sequence substitutions in the major mite allergens can markedly affect immunoglobulin E binding and T cell responses, but there are few studies on environmental isolates from Dermatophagoides pteronyssinus and none for D. farinae. OBJECTIVE: To determine the sequence variation of the group 1 and 2 allergens from environmental D. pteronyssinus and D. farinae. METHODS: RNA from each species was isolated from homes in Bangkok and the sequence of Der p 1, Der p 2, Der f 1, and Der f 2 determined from cDNA produced by high fidelity polymerase chain reactions. RESULTS: The enlarged data set revealed preferred amino acid substitutions in residues 19, 81, and 215 of Der p 1 as well as sporadic changes. Der p 2 showed frequent variations with clusters of amino acid substitutions, but the canonical Der p 2.0101 was not found in any of 17 sequences. Der f 2 showed variants with clusters of substitutions similar to Der p 2 but in different amino acid positions and without any structural concordance. Der f 1 in contrast to the other allergens had few amino acid sequence substitutions. CONCLUSIONS: The sequence information on variants provides data important for the optimal design of allergen formulations and useful for the genetic engineering and structure-function analyses of the major allergens.     

Hyposensitization in asthmatics with mPEG modified and unmodified house dust mite extract

Forty-six asthmatics with verified allergy to the house dust mite, D. pteronyssinus (Dp), participated in a double-blind study comparing the effect of 2 years' hyposensitization with two different Dp extracts. Two groups received either monomethoxypolyethylene glycol modified (mPEG) Dp extract or the corresponding non-modified extract, and a third group acted as controls receiving no injections. Medicine consumption, symptom scores, and peak expiratory flow (PEF) were recorded daily from September to December prior to and after 6 and 18 months of treatment. Changes were calculated choosing changes greater than or equal to 10% as relevant. In addition, patients were asked to give their direct assessment of the clinical effect at the end of the study. After 6 months, there was an improvement in symptoms + medication in 11/14 of Dp-treated, 6/17 of the mPEG-Dp group (P greater than 0.05) and 3/15 of openly treated controls. Few patients had changed in PEF. During the second year, several Dp-treated relapsed and some controls improved. At the end of the study the same improvement rate was seen in all groups. Similarly, the retrospective questionnaire data did not disclose any significant differences between groups after 2 years. In conclusion, hyposensitization with unmodified Dp extract seemed to have a favourable short-term effect on bronchial symptoms + medication in the majority of patients. When mainly on maintenance dose, the beneficial effect was reduced. The mPEG modification of the extract had reduced not only allergenicity but also the clinical effect of equal doses. Changes in medicine and symptom scores only partly correlated to retrospective assessment, thus stressing the problems in this kind of evaluation.     

Clinical improvement and immunological changes in atopic dermatitis patients undergoing subcutaneous immunotherapy with a house dust mite allergoid: a pilot study

BACKGROUND: House dust mites (HDMs) represent significant indoor allergen sources for patients with atopic dermatitis (AD). Subcutaneous allergen-specific immunotherapy (SCIT) has been shown to be successful in patients with allergic rhinitis and mild asthma and might represent an attractive therapeutic option for the long-term treatment of HDM sensitizations in AD patients. However, only a few studies have been conducted on the effectiveness of HDM SCIT in AD, resulting in controversial clinical results. Data on immunological changes induced by SCIT in AD patients are rare. OBJECTIVES: We performed an open pilot study to assess clinical changes and objective laboratory parameters and evaluate the benefit of HDM SCIT in 25 AD patients with IgE-mediated sensitization against HDM. METHODS: The severity of AD was evaluated by the severity scoring of atopic dermatitis system (SCORAD). Specific IgE and IgG4 against HDM and serum levels of TARC/CCL17, MDC/CCL22, IL-16, IL-4, IFN-gamma, IL-10 and TGF-beta1 were measured during SCIT. RESULTS: Subjective and objective SCORAD improved significantly within only 4 weeks of treatment. The level of the tolerogenic cytokine IL-10 increased, whereas CCL17 and IL-16 decreased in the sera of the patients during SCIT. Allergen specific IgE decreased, while IgG4 increased during SCIT. CONCLUSION: In this open-label pilot study, SCIT with an HDM extract in patients with AD led to a significant improvement of AD mirrored by a reduction of SCORAD as well as serological and immunological changes, which might serve as valuable parameters to estimate the therapeutic effect of SCIT.