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House dust mites in Bristol.

A survey of bedding in Bristol revealed reducing mite concentrations on children's toys, mattresses, overblankets and pillows. Dermatophagoides pteronyssinus was the commonest mite found. A vacuuming schedule was devised for removing mites so that a double blind trial could be carried out. Investigation suggests that even with expert utilization of efficient vacuum cleaners the clinical efficacy would be limited as the mites appear to rapidly migrate back into the bedding.

Allergens↗

Sensitivity to house dust mite and grass pollen in adults. Influence of the month of birth.

A comparison was made between the birth month of a control population and a sample of 240 adult patients with bronchial asthma or rhinitis and positive skin test towards house dust mite (Dermatophagoides pteronyssinus). The patients were born more frequently (P less than 0.005) in the summer and autumn months than in the other seasons. A similar comparison of 336 allergic patients with a positive skin test towards grass pollen (Phleum pratense) but no consistent seasonal preference in the birth months was revealed. This was possibly explained by the age of the patient group studied. The increased incidence of house-dust mite allergy in patients born in the months of May to September inclusive when house dust mites are supposed to be most abundant corresponds to a relative risk of 1.44. It is important to diminish the exposure to house dust mites in early childhood because exposure to allergens may influence the development of allergic disease in later life.

Adult↗

Circulating immune complexes in patients with house-dust-mite-sensitive bronchial asthma.

Sera from forty patients with house-dust-mite-sensitive bronchial asthma were examined for the presence of circulating immune complexes (CIC) by the sensitive and quantitative Clq solid-phase radioimmunoassay (Clq-SP) and a monoclonal rheumatoid factor solid-phase radioimmunoassay (mRF-SP). Compared to fifteen normal individuals, the asthmatic patients showed significantly higher mean values of Clq-reactive materials; however, there was no difference between the results from the patients treated by immunotherapy using Dermatophagoides farinae extract and those not so treated. Moreover, immune complexes in eight patients before and after immunotherapy showed that the amount of the complexes tend to decrease during immunotherapy. Furthermore, the presence of complexes had no relationship with the amounts of mite-specific IgG antibody. Similar results were also obtained in tests using mRF-SP. These data suggest that complexes in the sera of the house-dust-mite-sensitive asthmatic patients are not necessarily associated either with immunotherapy or with the mite-specific IgG antibodies.

Antigen-Antibody Complex↗

Reduction of house dust mite allergen levels in the home: use of the acaricide, pirimiphos methyl.

House dust mite sensitivity is very common in patients with bronchial asthma, yet dust mite avoidance frequently receives little attention in clinical management. It is likely that any reduction in allergen levels associated with routine cleaning is insufficient to allow clinical improvement. In the present study the acaricide pirimiphos methyl is shown to reduce the levels of Dermatophagoides pteronyssinus, antigen P1 in homes. Following a single application the level of antigen P1 in dust from carpets was reduced by up to 73% and by more than 50% in soft furnishings. Serial sampling showed a reduction for 6 weeks under conditions where carpets and chairs treated with solvent showed a progressive rise in allergen level. Furthermore the survival of mites in cultures or infested carpet segments was markedly inhibited, with antigen P1 accumulation reduced by greater than 90%. These results suggest major reductions in house dust mite allergen levels in the home can be achieved.

Allergens↗

Use of liquid nitrogen in the control of house dust mite populations.

Laboratory and field studies on the use of liquid nitrogen as an acaricide, combined with vacuum cleaning, demonstrated this inert freezing agent to be extremely efficient in reducing the number of live mites in mattresses when compared to vacuum cleaning only. Preliminary observations suggest this treatment does no physical damage to mattresses. Any strategy in the control of house dust mites should incorporate procedures both for the reduction of the mite population and the removal of the allergen pool of faecal pellets and dead mites.

Beds↗

Guanine and mite allergenicity in house dust.

Guanine is the major nitrogenous waste product in arachnids. It may serve as an indicator for allergenic mite faecal pellets. The present study assesses the correlation between the mit allergenicity of different house dust (HD) samples and their guanine content. Guanine content in HD samples was evaluated either by the Acarex-Test or determined quantitatively, especially for in-vitro methods. Using intradermal tests in selectively sensitized Dermatophagoides pteronyssinus (Dpt) patients, we evaluated the mite allergenicity of eight extracts obtained from HD samples with varying guanine contents. The weal area appeared to be proportional to the HD samples' guanine content. A three-fold increase in skin reactivity was observed when the guanine content varied from 0.06% to more than 1%. Patients allergic to HD but not to Dpt had no significant reactions with HD extract prepared from a guanine-rich HD sample. For eighteen different HD samples coupled to paper discs, the percentage Dpt RAST binding values obtained with a Dpt-selectively sensitized patient serum pool was correlated with the guanine concentrations of HD samples (P less than 0.01). The potencies of different HD extracts obtained from HD samples that varied in guanine content were also clearly distinguished on histamine-release titration curves. House dust obtained from homes at high altitude or from hospitals was generally guanine free. The HD sample guanine contents in homes where twelve Dpt-sensitized patients experienced symptoms were significantly higher than in homes where these patients were symptom free. The guanine colour test, a quick and easy technique, represents an important new development in indoor environmental investigations. It makes it possible to demonstrate mite faecal exposure and can be used to monitor the effectiveness of mite allergen avoidance measures.

Allergens↗

The subclass of IgG antibody in allergic disease: II. The IgG subclass of antibodies produced following natural exposure to dust mite and grass pollen in atopic and non-atopic individuals.

In an attempt to understand the role of the different IgG subclasses in allergic disease, we have studied the subclass of IgG antibody to dust mite (HDM) and grass pollen (GP) produced as a result of natural exposure. Studies were carried out on 40 atopic children and 100 atopic adults who had never received immunotherapy. Thirty-two non-atopic adult controls were also studied. The specificity of the assay for IgG antibodies to dust mite was confirmed by inhibition with the homologous extract but not mite culture medium or fetal calf serum. IgG1 antibodies to HDM could be detected in most atopics (94%) and non-atopics (97%), and similar results were obtained for GP (81% and 100%, respectively). IgG4 antibodies to HDM were detected in more atopics (66%) than non-atopics (53%) and the difference was more marked for GP (72% vs. 19%). While the levels of IgG1 antibodies were not significantly different in the two groups, the levels of IgG4 antibodies were much lower in the non-atopics (P less than 0.001, Mann-Whitney U-test). These data show that all subjects were capable of recognizing and mounting an IgG1 antibody response to these inhaled antigens. Atopic individuals differed from normal subjects in the frequency with which they made IgG4 antibodies in response to natural exposure to both dust mites and pollen.

Adult↗

Sequence analysis of cDNA coding for a major house dust mite allergen, Der f I.

A cDNA clone coding for Der f I, a major allergen from the house dust mite Dermatophagoides farinae has been isolated and sequenced. It codes for a putative 18-residue signal peptide, an 80-residue proenzyme region, and a 223-residue mature protein with a derived molecular weight of 25,191. The deduced amino-acid sequence shows significant homology to other cysteine proteases in the proregion as well as in the mature protein. Sequence alignment of the mature Der f I protein with the homologous allergen Der p I from the related mite D. pteronyssinus revealed a high degree of homology (81%) between the two proteins, as predicted by previous sequencing at the protein level. In particular, the residues comprising the active site of these enzymes and the cysteine residues were conserved. A potential N-glycosylation site was present at an equivalent position in both mite allergens. It is anticipated that the availability of recombinant Der f I will facilitate epitope mapping studies and studies of T-cell function in mite allergy by providing high levels of pure allergen.

Allergens↗

Immunoblot multi-allergen inhibition studies of allergenic cross-reactivity of the dust mites Lepidoglyphus destructor and Dermatophagoides pteronyssinus.

The allergenic similarity of the pyroglyphid mite D. pteronyssinus and the glycyphagid mite L. destructor was investigated with a new immunoblotting inhibition technique allowing simultaneous comparison of several allergens. Extracts of D. pteronyssinus and L. destructor were separated by SDS-PAGE and electroblotted to nitrocellulose (NC). A serum pool containing IgE specific to the major allergens in both mites was mixed with serially diluted extracts of D. pteronyssinus and L. destructor and incubated with the mite allergens of NC. The inhibition of the IgE binding to NC was evaluated by densitometric scanning and percentage inhibition was calculated. The IgE antibodies to the 25-kD component in D. pteronyssinus, were inhibited to the same degree by extracts of D. pteronyssinus and L. destructor. Another major allergen component in D. pteronyssinus (16 kD) was also inhibited by L. destructor extract but to a lesser degree: 400 times more of the heterologous than of the homologous extract was needed for 50% inhibition. To produce 50% of heterologous inhibition of the two major allergen components at 15 and 53 kD of L. destructor, 2000 and 10,000 times more respectively, of D. pteronyssinus than of L. destructor extract were needed. Two minor allergen components of L. destructor showed some cross-reactivity with D. pteronyssinus. However, L. destructor was a stronger inhibitor of D. pteronyssinus than vice versa, probably because the sera were obtained from persons more sensitized to L. destructor than to D. pteronyssinus.

Allergens↗

Comparison of allergic responses to dust mites in U.K. bakery workers and Swedish farmers.

The IgE RAST response to Dermatophagoides pteronyssinus and four storage mites (Lepidoglyphus destructor, Tyrophagus putrescentiae, Glycyphaghus domesticus, and Acarus siro) was examined in 251 U.K. bakery workers and compared with that previously found in 440 Swedish farmers. Storage mites are found commonly in stored hay and grain so both these groups potentially encounter them in their work. In neither group of workers was a positive RAST (greater than or equal to 0.35 PRU) to D. pteronyssinus correlated with a positive RAST to a single storage mite. As in the Swedish farmers, significant though not strong correlations were found in the U.K. bakers between positive RAST responses to G. domesticus and L. destructor and to T. putrescentiae and L. destructor (P less than 0.05). Homologous and heterologous RAST inhibition studies showed there was low cross-reactivity between storage mites and D. pteronyssinus. L. destructor showed the least inhibition by the other antigens, suggesting it possessed the fewest common allergens. The most important difference in the IgE responses between the two groups was the much higher response to D. pteronyssinus in the U.K. bakers, which was not found in the Swedish farmers whose highest IgE response was to L. destructor.

Agriculture↗

Localization of allergens in the domestic mite Lepidoglyphus destructor.

The non-pyroglyphid domestic mite Lepidoglyphus destructor is a major source of allergen causing respiratory symptoms in farming environments. This study is the first to focus on the localization of the allergens in the non-pyroglyphid domestic mite Lepidoglyphus destructor. Cryostat-cut sections of L. destructor mite bodies and faecal pellets were probed with one of three mouse monoclonal antibodies (MoAbs) raised against L. destructor or with patient sera, and stained with immunoperoxidase. Eight sera were RAST-positive to L. destructor and L. destructor faecal pellets. These eight RAST-positive patient sera labelled the wall of the L. destructor gut and two of them also the faecal pellets. The MoAbs additionally labelled parts of the exoskeleton and reacted with a majority of the faecal pellets. Cryostat-cut sections of Dermatophagoides pteronyssinus mite bodies and faecal pellets were probed with L. destructor MoAbs, which resulted in only slight staining of a few faecal pellets. The results suggest that at least one L. destructor allergen is associated with digestion.

Adult↗

Mite antigen in house dust: relationship with different housing characteristics in The Netherlands.

As part of a case-controlled study on the relationship between home dampness and respiratory symptoms of children, the concentration of the major allergen of Dermatophagoides pteronyssinus (Der p I) in floor dust and mattress dust in 516 dwellings in the Netherlands was measured. A checklist, completed by the investigators, was used to obtain information on home and occupant characteristics, which may have an impact on the Der p I concentration in house dust. The geometric mean mite antigen concentrations were 2370 ng Der p I/g floor dust for the living room, 2201 ng Der p I/g floor dust for the bedroom and 5075 ng Der p I/g mattress dust. In 86% of the houses the maximum concentration was higher than 2000 ng Der p I/g dust, that is regarded as representing a risk for genetically predisposed individuals for the development of specific IgE to house dust mite allergen. In 55% of the houses the maximum concentration exceeded 10,000 ng Der p I/g dust, regarded as a risk factor for acute attacks of asthma for mite allergic patients. The Der p I concentrations in dust from carpeted floors were six to 14 times higher than in dust from floors with a smooth floor covering. Higher Der p I concentrations in floor dust were also significantly associated with increasing age of the dwelling and of the floor covering, with an increasing number of occupants, and with the absence of floor insulation. For mattress dust, the age of the mattress, the presence of an outer cavity wall and mechanical ventilation were important factors. Older mattresses had higher levels, and mattress dust from bedrooms with solid brick outer walls had higher levels than that from bedrooms with outer cavity walls. Mattresses in homes with continuous mechanical ventilation had almost twice lower levels than mattresses in homes with natural ventilation. There was a tendency towards higher Der p I concentrations in dust in homes with reported or observed signs of dampness. The Der p I concentrations in dust from carpeted bedroom floors and mattresses were positively associated with the average relative humidity in the bedroom over a period of 3-6 weeks in a subset of the homes where relative humidity was measured. Similar results were obtained using the concentrations of Der p I in ng/m2 instead of ng/g dust. The results obtained in this study are of importance for planning and evaluating allergen avoidance measures advised to mite allergic patients.

Adult↗

Estimation of Der p and Der f I quantities in the reference preparations of Dermatophagoides mite extracts.

A monoclonal antibody-based enzyme-linked immunosorbent assay (MoAb-ELISA) was developed to measure the major Dermatophagoides mite allergens, Der p I and Der f I. The assay was highly species-specific and sensitive. Using this assay system, the absolute mass unit of Der p I and Der f I in the reference preparations of the extracts was estimated. The primary standards used were the purified Der p I and Der f I preparations. The reference preparations of the D. pteronyssinus and D. farinae extracts (92-Dp and 92-Df), which had been prepared from the same amount of mite bodies of both species, were found to contain the same levels of the Der I allergens, 10.1 micrograms/ml of Der p I and 10.0 micrograms/ml of Der f I, respectively. A histamine release assay with leucocytes from mite-allergic donors showed that the total allergenic potency of 92-Dp and 92-Df was comparable. This results indicates that the estimated Der I levels in these extracts seem to be valid, at least, in the balance between the two species, although further comparisons of the absolute quantities by several different laboratories are needed. The Der I levels in the WHO/IUIS international reference preparation of D. pteronyssinus and the CBER standard mite extracts, E4-Dp and E5-Df, were also estimated using this assay system. They were found to contain 4.4 micrograms/vial and 13.3 micrograms/ml of Der p I and 9.5 micrograms/ml of Der f I, respectively.

Allergens↗

Identification of allergens from the mite Blomia tropicalis.

In some tropical areas the mite Blomia tropicalis is a clinically important allergenic component of house dust, inducing specific IgE immune response in patients with allergic respiratory diseases such as asthma and rhinitis. The identification of allergens of this mite is necessary to obtain appropriate reagents for diagnostic and treatment procedures. We carried out this study using immunoblotting to detect the allergens of B. tropicalis. Our results demonstrate that this mite has one major allergen (11-13 kDa) and three other important allergens with about 50% binding (64, 36 and 33 kDa). Therefore, B. tropicalis should be regarded as an important source of allergens in the house dust in tropical areas, besides those derived from other mites.

Adolescent↗

The decay of house dust mite allergens, Der p I and Der p II, under natural conditions.

BACKGROUND: Fluctuations in the level of mite allergens in domestic house dust are the result of changes in the balance between synthesis, removal and decay. Purely physical forces as well as enzymatic degradation, mediated by house dust inhabiting microbes, may contribute to the decay of allergens in domestic dust. Knowledge about the speed of decay is essential for an understanding of the dynamics of allergen levels. OBJECTIVE: The present study is a quantitative assessment of the speed of decay at nine combinations of temperature (15 degrees C, 20 degrees C and 25 degrees C) and relative humidity (33%, 55% and 75%). METHODS: Samples of mite infested material of an old rug were stored at these temperature/relative humidity-combinations for 6, 12 or 18 months, after the mites were killed by either a freezing treatment or an acaricide (lindane). The microbes living in the rug presumably survive these treatments. Concentrations of Der p I and Der p II + Der f II, in extracts of the rug material, were measured by radio immunoassay. RESULTS: No significant changes in the levels of der p I and Der p II + Der f II, could be detected even after 1 1/2 year at a high temperature and humidity. CONCLUSION: These findings indicate that mite allergens can be extremely stable under normal domestic circumstances.

Allergens↗

Localization of major allergens in the dust mite Lepidoglyphus destructor with confocal laser scanning microscopy.

The dust mite Lepidoglyphus destructor is the dominating source of allergens giving rise to asthma and rhinitis among farmers. In a previous study of the localization of allergens in L. destructor we demonstrated that the 39 kDa allergen is associated with digestion. Here we describe the localization of the principal 15 kDa allergen and the high molecular weight allergen complex (79 and 93 kDa) in L. destructor with confocal laser scanning microscopy (CLSM). Cryostat-cut sections of mite bodies and faecal pellets were probed with mouse monoclonal antibodies (MoAbs) raised against the allergens. The 15 kDa allergen disclosed labelling of the mite body and most of the faecal pellets but left the exoskeleton unlabelled. The binding was widespread, and most intense in the mouth region. However, some staining was also observed around the gastrointestinal tract. In contrast, the 79 and 93 kDa allergen complex stained the exoskeleton and the front part of the mite. Interestingly, we detected no labelling of the faecal pellets with the MoAb against the 79/93 kDa allergen. The study indicates that the 15 kDa allergen is associated with the digestive tract whereas the function of the 79 and 93 kDa allergen complex remains to be elucidated.

Allergens↗

A combined approach to reduce mite allergen in the bedroom.

BACKGROUND: Asthma is a common chronic disease of childhood. House dust mite (HDM) are known to be a major source of allergen affecting atopic asthmatics. No single control method has been demonstrated to consistently reduce asthma. OBJECTIVE: We investigated the effect of a combination of two methods of HDM allergen control on HDM allergen content in the bedding and carpets of asthmatic children. METHODS: This was a double-blind placebo-controlled trial treating the bedrooms of 56 mite-sensitive asthmatic children. The carpet and the mattress, duvet and pillows (bedding) in the bedroom of children of the active group were treated with the acaricide Acarosan (benzyl benzoate). The bedding was then encased in vapour permeable waterproof fabric (Intervent--cotton coated with polyurethane) for 24 weeks. The carpet and bedding of the control group were treated with placebo and the bedding encased in cotton covers for 24 weeks. Dust samples were collected from all these items in a standard manner at regular intervals and Der p I content analysed. RESULTS: The group with active treatment had a median reduction of 480 ng (100%) in mite allergen from the mattress vs 215 ng (53%) reduction in placebo-treated group by 6 weeks. The Der p I content of the active group's bedding was always less than the placebo group after treatment (P < 0.01). The acaricide applied to the carpets or inside the mattress covers was ineffective in reducing allergen content. CONCLUSION: This study confirms the effectiveness of encasing covers in reducing the mite allergen exposure but indicates there is no further advantage in applying acaricide simultaneously.

Allergens↗

Absence of continuous epitopes in the house dust mite major allergens Der p I from Dermatophagoides pteronyssinus and Der f I from Dermatophagoides farinae.

BACKGROUND: The house dust mite has been shown to be an important source of domestic allergens associated with immediate hypersensitivities. The Group I mite allergens Der p I from Dermatophagoides pteronyssinus and Der f I from D. farinae display extensive amino acid sequence homology and have similarities with cysteine protease enzymes. OBJECTIVE: The availability of the complete amino acid sequences for these allergens allowed us to search for the allergic determinants within these molecules. The aim of the present investigation was to identify any continuous IgE-binding epitopes within these amino acid sequences. We also sought to test the validity of previously reported Der p I peptide epitope sequences. METHODS: In order to identify any continuous IgE epitopes, the amino acid sequences of Der p I and Der f I were synthesized as decapeptides overlapping in sequence and coupled to plastic pins. The specific IgE-binding capacity of these peptides was assayed using an enzyme-linked biotin-streptavidin procedure and sera from patients known to be sensitive to these allergens. Previously reported Der p I peptide epitopes were synthesized as free peptides and tested for their ability to inhibit specific IgE binding to allergen extract discs. RESULTS: None of the pin-coupled Der p I or Der f I peptides was found by the continuous epitope mapping procedure to bind significantly to specific IgE in the sera of hypersensitive patients. The previously reported Der p I peptide epitopes did not inhibit specific IgE binding to mite extract discs. CONCLUSION: The specific IgE binding epitopes of the house dust mite allergens Der p I and Der f I are discontinuous in nature.

Allergens↗