Glutathione S-transferase a major allergen of the house dust mite, Dermatophagoides pteronyssinus

Cutaneous electrogastrography (EGG) allows the measurement of gastric electrical activity. An association of EGG with gastrointestinal motility disorders has been shown. Abnormalities of electrical rate or rhythm are accepted as the most important parameters in EGG. However, the reliability of the magnitude of electrical amplitude in the assessment of motility is discussed controversially. Therefore in a prospective study we investigated the relation between amplitude and antral contractions by means of ultrasonography. 8 healthy volunteers (4 men, 4 women, 24-31 years) ingested 400 ml carbonated mineral water after an overnight fast at two separate study days. Over a period of 10 min preprandial and 10 min postprandial small and intense antral contractions were measured employing sagittal antral planimetry. Simultaneous amplitudes were determined during contractions and at 1 min intervals (average amplitude) by cutaneous electrogastrography. Data were analyzed by Wilcoxon's rank sum test and Spearman rank correlation test. The coefficient of variation of the postprandial/preprandial amplitude ratio was nearly two times greater between subjects than between recordings in the same subject, which reflects a moderate intraindividual reproducibility. We found a significant increase in the average amplitude postprandially (p < 0.05). Although postprandial contractions (n = 243) predominated preprandial contractions (n = 127) significantly (p = 0.02), no significant correlation between the number of contractions and the average amplitude existed (R = 0.1; p = 0.7). Moreover the average amplitude did not differ from amplitudes during intense and small contractions significantly (p = 0.7; p = 0.1). The magnitude of the amplitude measured by EGG does not correlate with the mechanical gastral activity significantly.(ABSTRACT TRUNCATED AT 250 WORDS)     

House dust mite allergen accumulation on sheepskins

AIM: Sheepskins, which are often used as infant bedding in New Zealand, are known to harbour large quantities of house dust mites and their allergens. In this study we determined the rate of accumulation of the house dust mite allergen, Der p 1 on new sheepskins, and the effects of washing and dry cleaning on its removal. METHODS: New sheepskins were placed on living-room floors (n = 6) and mattresses (n = 6) in six domestic dwellings in Wellington. Sheepskin dust samples were collected by vacuuming before placement, and at two, four and six weeks, after which they were warm-washed. They were replaced for a further six weeks, then dry-cleaned. Dust samples were collected before and after washing and dry-cleaning, and from mattresses and living-room floors. Dust samples were analysed for Der p 1 by double monoclonal antibody ELISA. RESULTS: Der p 1 levels rapidly increased in sheepskins placed on living-room floors and mattresses to a geometric mean level (range) of 9.0 micrograms/g (1.1-102.2) and 29.4 micrograms/g (5.3-131.1) at six weeks, respectively. After warm-water washing and replacement for a further six weeks, these levels were higher at 75.5 micrograms/g (50.1-260.4) and 31.9 micrograms/g (11.1-75.2), respectively. Sheepskin Der p 1 accumulation correlated with mattress (r = 0.78), and living-room floor Der p 1 levels (r = 0.94). Warm water washing and dry-cleaning reduced sheepskins Der p 1 levels by a mean of 79.2% and 95.3%, respectively. CONCLUSIONS: Sheepskins rapidly accumulate house dust mite allergens from the domestic environment. Due to very high levels and rapid accumulation of Der p 1, sheepskins as infant bedding should be discouraged for infants at risk of sensitisation to house dust mites. If they are to be used then it is important that they are washed or dry-cleaned regularly.     

Tertiary structure of the major house dust mite allergen Der p 2: sequential and structural homologies

Sensitization to indoor allergens, especially those of the house dust mite, is strongly correlated with the development of asthma. We report the tertiary structure of the major house dust mite allergen, Der p 2, determined by NMR methods. The structure of Der p 2 is a beta-barrel and is composed of two three-stranded antiparallel beta-pleated sheets. This arrangement of beta-strands is similar to the immunoglobulin fold with respect to the orientation of the two sheets and the interactions of the strands. However, the three-dimensional structure of Der p 2 aligns equivalently with a number of proteins from different families within the immunoglobulin superfamily. The structural homology with the highest significance score from analysis by DALI is to Der f 2. Although Der p 2 and Der f 2 are 87% identical in amino acid sequence, they align in three dimensions rather poorly (4.85 A RMSD; Z-score, 8.58). This unexpected finding is likely due to the different solution conditions used during structure determination by NMR for both proteins. While the structural comparisons did not elucidate a clear homologue for the function of Der p 2 in mites, we report that Der p 2 is sequentially homologous to esr16. This is a protein from moths that is expressed coincident with molting. Thus, this homology has important ramifications for the study of mite allergy. The structure of Der p 2 provides a useful tool in the design of recombinant immunotherapeutics for the group 2 allergens.     

Cloning and characterization of a major allergen of the house dust mite, Dermatophagoides pteronyssinus, homologous with glutathione S-transferase

A major allergen of the house dust mite, Dermatophagoides pteronyssinus, has been identified and characterized from a lambda gt11 cDNA library of the mite. IgE antibodies from the sera of allergic patients that recognise the cloned polypeptide bind to an approximately 26 kDa polypeptide on a Western blot of reduced mite polypeptides. Nucleotides sequencing of the clone revealed a 219 amino acid open reading frame encoding a protein with a derived molecular mass of 25,589 Da and a pI of 6.3. Comparison of the deduced amino acid sequence with amino acid sequence databanks revealed a strong homology with glutathione S-transferases. The nucleotide sequence of the clone displayed a strong homology with the active glutathione binding site of glutathione transferases and contained all but one of the 19 positionally conserved amino acid residues found in glutathione transferases. The cloned polypeptide was expressed in Escherichia coli and affinity-purified on glutathione agarose.     

Induction of atopic dermatitis by inhalation of house dust mite

BACKGROUND: The pathogenetic role of house dust mite in atopic dermatitis remains controversial. Recent studies have shown that intensive epicutaneous contact of house dust mite allergen with premanipulated skin may induce dermatitis. It is, however, uncertain whether such conditions are met during natural contact with house dust mite. In the past, allergen inhalation has been suggested to induce exacerbation of atopic dermatitis. The aim of this study was to investigate whether dermatitis could be induced in patients with atopic dermatitis by inhalation of house dust mite. METHODS: Twenty patients with atopic dermatitis underwent bronchial provocations with house dust mite. Challenge tests were performed with four concentrations of a standardized house dust mite extract in a double-blind, randomized, placebo-controlled fashion. Spirometry was performed, and FEV1 was measured before and after each challenge dose. Changes in severity or localization of itching or erythema were recorded. RESULTS: In nine of 20 patients with atopic dermatitis bronchial challenge with house dust mite induced unequivocal skin symptoms after 1.5 to 17 hours. Pruritic erythematous lesions on noninvolved sites together with exacerbations of existing lesions were seen in three patients. Three patients had an exacerbation only, and three other patients had new lesions only. In eight of nine patients with house dust mite inhalation-induced dermatitis, skin symptoms were preceded by an early bronchial reaction. All patients with house dust mite-induced dermatitis had a history of asthma, and as a group they had a higher mean blood total IgE level compared with the "negative skin responders." One patient had pruritic erythema on the placebo challenge day, without a preceding bronchoconstrictive reaction. The number of patients who had a skin response on the house dust mite challenge day was significantly higher than the number of patients who had a skin response on the placebo day (p = 0.011 [Prescott's test]). CONCLUSIONS: The respiratory route may be relevant in the induction and exacerbation of dermatitis in a subset of patients with atopic dermatitis who have early bronchial reactions after house dust mite inhalation, a history of asthma, and an elevated blood total IgE level. Furthermore, these findings suggest a possible causal relationship between bronchial reactions and skin reactions.     

Phospholipid metabolism in European house dust mite (Acari: Pyroglyphidae)

Phospholipid metabolism was studied in European house dust mite, Dermatophagoides pteronyssinus (Trouessart). Analysis of phospholipid metabolites in mite extracts suggested the presence of PLaseA1, PLaseA2, and PLaseC, which have important physiological roles in phospholipid metabolism. The optimum pH of PLaseA1 and PLaseA2 was 8 and 9, and the optimum temperature of PLaseA1 was 45 degrees C; the activity of PLaseA2 was not affected by temperature. The activity of PLaseA1 at pH 8 and 37 degrees C was 5 times higher than that of PLaseA2 in D. pteronyssinus. The optimum temperatures of diacylglycerol production were 25 and 45 degrees C.     

Specific IgA and IgG antibodies to house dust mite Dermatophagoides farinae in nasal secretions

As far as we know, IgA and IgG antibodies to purified house dust mite allergens Der fI and Der fII in nasal secretions have never been documented. Therefore, we determined specific IgA, SIgA and IgG antibodies (abs) to crude extract of D. farinae and its purified allergens Der fI and der f II in nasal secretions collected by aspiration from 34 normal subjects, 25 untreated nasal allergic patients and 28 treated nasal allergic patients on parenteral immunotherapy by means of an avidin-biotin ELISA. The following results were obtained. (1) The specific IgA, SIgA and IgG abs to each of the three kinds of allergens correlated with each other. The groups of patients with nasal allergy (both treated and untreated) showed higher levels of specific IgA, SIgA and IgG abs to the allergens than the normal group. (2) In the group of treated patients, the levels of specific abs were not correlated with the clinical improvement of symptoms or the degree of response to nasal challenge. (3) The treated patients failed to show significantly higher levels of abs in nasal secretions than the untreated patients. (4) The specific IgA and SIgA abs in nasal secretions seemed to be predominantly produced locally, and IgG abs might be transudated from the circulation.     

Adhesion molecules in atopic dermatitis: patch tests elicited by house dust mite

Different T-helper subsets, which are characterized by the secretion of distinct cytokines (Th1, Th2), have been found in house dust mite-exposed skin of sensitized individuals and in nickel-specific T lymphocytes from nickel contact allergic and non-allergic individuals. In order to evaluate the role which adhesion molecules may play in the homing of different T-cell subsets into allergen-exposed skin of atopic and normal individuals, we compared the expression pattern of adhesion molecules in patch test reactions to house dust mite antigen (D.pt.), nickel sulfate (Ni) and the irritant anthralin. Biopsies were taken at various time points after application of these agents and studied by immuncytochemistry. To exclude an endogenous difference in adhesion molecule expression in atopic and non-atopic skin, sequential biopsies from Ni patch tests of 2 normal individuals were also included in this study. The expression of E-selectin, P-selectin, CD31, VCAM-1 and ICAM-1 on endothelial cells and other cells in the skin was quantified by microscopic evaluation. Skin homing T cells were also quantified using antibodies to CD3, CD4, CD8, UCHL-1, L-selectin and the cutaneous lymphocyte antigen (CLA). Independent of the eliciting substance, all lesions showed an upregulation of all adhesion molecules tested, with the exception of CD62. The appearance of E-selectin and an increase in ICAM-1 and VCAM-1 expression were first observed at 12 h after application of the various agents. In parallel, the number of CLA+ and L-selectin+ lymphocytes increased steadily. No principle differences could be established between the various types of skin reactions in atopic individuals, nor did the skin of patients with AD differ from normal controls. Our results provide evidence that differential expression of adhesion molecules does not play a major part in observed differential homing of Th1 and Th2-cell subsets into patch test sites provoked by house dust mite and nickel sulfate in atopic and non-atopic individuals.     

Effect of a bed covering system in children with asthma and house dust mite hypersensitivity

Allergen avoidance is regarded as an important approach to management of atopic asthma. The effect of Intervent bed covering systems on house dust mite (HDM) allergen exposure, asthma symptoms and markers of inflammation was investigated in 31 HDM sensitive asthmatic children. Dust concentrations of Dermatophagoides pteronyssinus allergen 1 (Der p 1) were monitored before and after covering the mattress, duvet and pillow with active and placebo covers for 3 months, in a single-blind, cross-over trial. Twice daily peak expiratory flow rate (PEFR), daily symptom scores and treatment schedule were recorded. Bronchial hyperresponsiveness was monitored by histamine challenge (provocative concentration of histamine causing a 20% fall in forced expiratory volume in one second (PC20)), and inflammation by measuring eosinophil cationic protein (ECP), eosinophil protein X (EPX), eosinophil peroxidase (EPO), and soluble interleukin-2 receptor (sIL-2R) in serum. There was a significant reduction in Der p 1 when the mattress, duvet and pillow were covered with the active bedding. There was no significant improvement in symptoms of asthma, PEFR, bronchodilator usage of PC20. Also, ECP, EPX, sIL-2R concentrations did not change for either treatment. EPO concentrations were significantly lower in the active compared to the placebo period. The active bed covers reduced retrievable Dermatophagoides pteronyssinus allergen 1 (Der p 1) from the bedding, with short term clinical benefit.     

Prediction of murine MHC class I epitopes in a major house dust mite allergen and induction of T1-type CD8+ T cell responses

The group I (Der p 1) allergen of Dermatophagoides pteronyssinus (house dust mite, HDM) contains several T helper (Th) epitopes recognized by C57BL/6 mice, with the peptide (111-139) containing a dominant MHC class II-restricted epitope (113-127). Since CD8+ T cells are thought to play a role in the regulation of allergic disease, we examined the Der p 1 sequence for potential MHC class I-binding motifs and observed that residues 111-119 (FGISNYCQI) contain motifs for H-2Db and Kb. Furthermore, immunization of C57BL/6 mice with unadjuvanted Ty virus-like particles (VLP) carrying Der p 1 (111-139), a method known to induce murine cytotoxic T lymphocyte (CTL) responses, primed Der p 1 (111-119)-specific Db-restricted CTL which produce high levels of IFN-gamma and low levels of IL-5 and IL-6 in vitro (T1-type CTL). VLP carrying the minimal epitope (FGISNYCQI) also induced a CTL response following immunization without adjuvant by various routes. Der p 1 (111-139)-VLP adjuvanted with alum did not prime CTL in C57BL/6 mice but were found to prime Th1-type CD4+ T cells that recognize the overlapping peptide (113-127) and native protein. The ability to successfully predict allergen-specific CD8+ T cell epitopes and prime CD8+ and/or CD4+ T cell responses provides an opportunity to dissect the relative roles of these T cells in the regulation of allergic responses and may offer therapeutic potential for reprogramming Th2-type allergic responses.     

B cell mitogenic activity of house dust mite, Dermatophagoides farinae, antigens

Hong Kong, Taiwan, Singapore, and Malaysia are initiating health care reform to meet the changing demands of populations with improved socioeconomic status and access to modern technologies and who are living longer than in previous generations. Hong Kong, in particular, is facing a unique set of circumstances as its people prepare for the transition in 1997 from a British colony to a Special Administrative Region of China. While spending only 4% of its gross domestic product on health care, it has a large and regulated public hospital system for most inpatient medical care and a separate, loosely regulated private health care system for most outpatient medical care. In 1993 the Secretary for Health and Welfare of Hong Kong initiated a year-long process to debate the pros and cons of 5 fundamental programs for health care reform. After a year of open consultation, options were chosen. We describe the Hong Kong health care system, the fundamental changes that have been adopted, and lessons for reformers in the United States.     

Inactivation of heamolytic complement by house dust allergen in the serum of children with atopic diseases

Two experiments are presented in which lateralization of monaural, auditory input affected the evaluation of verbal passages. In Experiment 1, subjects listened to taped passages that consisted of three levels (positive, neutral, and negative) of tone of voice crossed with three similar levels of content or meaning, making a total of nine different tone/content combinations. Subjects rated these passages as positive, neutral, or negative, and although they were not instructed about which cues (tone or content) to use, 29 of 36 subjects who listened on the left ear primarily used the tone-of-voice cues to rate the passages, whereas 21 of 36 subjects who listened on the right ear used the content cues (p less than .01). In Experiment 2, subjects were asked to make objective ratings of both tone of voice and content. Subjects who listened on the right ear were more accurate in rating both cues. The lateral differences in evaluating the auditory stimuli are discussed in the framework of the left hemisphere's specialization for analytic processing of objective information and the right hemisphere's specialization for the holistic processing of subjective or emotional information. However, each hemisphere's specialization for tone and content cues can be greatly affected by the demands of the experimental task.     

Inhibition of human T-cell responses to house dust mite allergens by a T-cell receptor peptide

Recent analysis of the usage of T-cell receptor (TcR) beta chain variable region (V beta) gene elements by house dust mite (HDM)-reactive T cells from an atopic donor suggested that TcR-V beta 3 gene products may form a major component of the human T-cell repertoire reactive to this common allergen. In this study a peptide analog of the TcR-V beta 3 complementarity determining region 2 (CDR2) is shown to inhibit the polyclonal human T-cell response to HDM; this effect is specific because inhibition is dependent on the presence of V beta 3 + T cells. This experimental approach has been used to determine whether the pattern seen in T-cell clones derived from one atopic donor reflects TcR-V beta usage in the polyclonal response to allergen in the general population. Inhibition of more than 50% of the polyclonal response to allergen by V beta 3-CDR2 peptide was observed in 16 of 21 donors tested, suggesting that TcR-V beta 3 gene usage may form a major component of the human HDM repertoire and as such offer a suitable target for T cell-directed specific immunotherapy in HDM-allergic individuals. Depletion of CD8+ T cells abolishes peptide-mediated inhibition of CD4+ T-cell proliferation to HDM, suggesting that induction of a CD8+ regulatory T-cell subset by the CDR2 peptide may modulate HDM-specific allergic T-cell responses.     

Production of a mouse/human chimeric IgE monoclonal antibody to the house dust mite allergen Der p 2 and its use for the absolute quantification of allergen-specific IgE

1. We evaluated the antianginal effects of YM430 in several experimental models in vitro and in vivo. 2. In isolated dog coronary artery, YM430 (10(-8)-10(-6) M) inhibited 3,4-diaminopyridine-induced rhythmic contractions with an IC50 value of 59.2 nM. 3. In anesthetized rats, YM430 (10-100 mg/kg PO) inhibited arginine vasopressin-induced ST-segment depression with an IC50 value of 36.6 mg/kg PO. 4. In anesthetized dogs, YM430 (0.3 mg/kg IV) significantly inhibited ST-segment elevation induced by coronary artery occlusion. 5. These findings suggest that YM430 may be of value in the treatment of various types of angina pectoris such as variant and stable angina.     

Higher asthma occurrence in an urban than a suburban area: role of house dust mite skin allergy

Understanding of geographical differences in asthma prevalence may be helpful in explaining recent increases in the occurrence of asthma. We wondered whether differences in allergic sensitization or other factors could explain differences in reported occurrence of asthma between an urban centre and a neighbouring suburban area. From the European Community Respiratory Health Survey (ECRHS) questionnaire, responses on asthma symptoms and risk factors and results of 11 skin allergy tests were available from 656 young adults living in urban or south suburban Antwerp, Belgium. Answers to five asthma questions were selected as dependent variables, and eight personal or environmental risk factors, as well as house dust mite (HDM) allergy, as independent variables. The effect of each independent variable on the association of asthma variables with area was assessed. Prior asthma diagnosis, present asthma symptoms, the selected risk factors and HDM allergy were all more frequently recorded in urban Antwerp. Difference in HDM allergy accounted for most of the difference in prior (mostly childhood) asthma diagnosis, since correction for it decreased the odds ratio from 2.10 to 1.65. On the contrary, the regional differences in recent asthma symptoms were not explained by HDM allergy differences nor by any other factor under study. This urban-suburban comparison indicated that house dust mite allergy is a major determinant of prior (childhood) asthma, whereas factors contributing to higher urban prevalence of present asthma symptoms could not be identified. Furthermore, our results indicate that it may be inappropriate to combine data from neighbouring areas, when their similarity has not been verified.     

Epithelial repair in asthma. Do the benefits of house dust mite avoidance result from proteinase avoidance?

OBJECTIVE: To describe the timing of recovery of lung function after severe acute hypoxemic respiratory failure (AHRF) in children. DESIGN: A serial observational follow-up study of clinical and lung function measurements up to 53 months after acute illness. SETTING: University pediatric intensive care unit in a national children's hospital. PATIENTS: Five critically ill children aged 5-14 years. INTERVENTIONS: None RESULTS: Clinical recovery: each patient required a 3-5 month convalescence before being able to attend full-time school because of lethargy and dyspnea. All patients developed wheeze 3-12 months after illness and four received long-term bronchodilator therapy. Lung function recovery: for both the forced vital capacity (FVC) and forced vital capacity in the first second (FEV1) four patients had abnormally low values, regaining only 60-70% of predicted values for their height and sex, and all of this improvement had occurred by 6-12 months after illness. Beyond this interval, patients remained on their same FVC and FEV1 centile. FEV1/FVC ratios were consistently within the normal range, indicating a predominantly restrictive defect. Changes in peak expiratory flow exhibited a time course of improvement similar to the other lung function tests. CONCLUSION: In children, pulmonary recovery after severe AHRF may occur for 6-12 months. A 1-year follow-up could offer a rational single point for assessment of outcome and long-term counselling of child and parents.     

Measurement of airborne mite allergen exposure in individual subjects

To evaluate the extent of personal exposure to airborne mite allergens, subjects were asked to carry a personal air sampler when in their houses. The level of Der 1 allergen trapped by the sampler was measured with a highly sensitive immunoassay. There were great variations in airborne Der 1 exposure in each subject. When used bedding was replaced with new allergen-free bedding, we detected a decrease in the allergen level. The use of new bedding seems to be an effective measure for reducing airborne mite allergen exposure.     

Hypersensitivity to mites and house dust

The paper deals with the current data on hypersensitivity to house dust allergens, particularly to house dust mite allergens. Hypersensitivity to house dust mites is more prevalent than hypersensitivity to any other allergen. Ecologic factors for mite survival are cited, particularly emphasizing the importance of air humidity. Over the last 50 years there have been several changes in construction and furnishing of houses in developed countries which fostered mite proliferation. Mite allergens are identified and cloned, monoclonal antibodies to these allergens are produced. There are methods for estimating mite allergen concentration in homes which made possible to investigate the relationship between mite allergen exposure and development of bronchial hyperreactivity. Various methods for mite allergen avoidance are developed and their efficacy is under investigation.     

Infant respiratory symptoms in relation to mite allergen exposure

The relationship between the Dermatophagoides pteronyssinus (Der p) I content of house dust and the respiratory symptoms reported for young infants was studied. One hundred and four infants, aged 3-15 months, were selected during July-September 1993 through the Dutch postnatal health care service, using a short screening questionnaire to identify mothers with respiratory allergy to house dust and/or pets. Forty-eight were selected from this group of mothers ("high risk" infants) and 56 infants were selected when neither of the parents reported allergy or chronic respiratory symptoms ("low risk" infants). All homes were visited in October 1993. Dust samples were collected from the infant's mattress and from other places in the home, and the Der p I content was measured in dust extracts. The results indicate that on more than half of the mattresses, the Der p I level was over 2,000 ng.g-1, the level suggested to be associated with an increased risk of sensitization. Information on respiratory symptoms ("wheeze" and "prolonged cough") experienced since birth was obtained by questionnaire from one of the parents on the dust sampling day. The occurrence of respiratory symptoms in the infants appeared to be positively related to the Der p I concentration of the dust. Although no objective measurements of respiratory symptoms were available, the results of this study suggest that exposure to mite allergen in early life may lead to respiratory symptoms that are suggestive of airway obstruction in the first year of life.