Patterns of IgE sensitization in house dust mite-allergic patients: implications for allergen immunotherapy.

BACKGROUND: Understanding patterns of IgE sensitization in Dermatophagoides-allergic patients living in various geographical areas is necessary to design a product suitable for worldwide allergen immunotherapy (AIT). METHODS: Using a HIFI Allergy customized microarray assay, IgEs specific for 12 purified allergens from Dermatophagoides pteronyssinus or D. farinae were assessed in sera from 1302 house dust mite (HDM)-allergic patients living in various areas. Comprehensive mass spectrometric (MS) analyses were conducted to characterize HDM extracts, as well as purified bodies and feces. RESULTS: Patterns of IgE reactivity to HDM allergens are comparable in all cohorts of patients analyzed, encompassing adults and 5- to 17-year-old children, as well as American, Canadian, European, and Japanese patients. Overall, >70% and >80% of HDM-allergic patients are sensitized to group 1 and group 2 allergens, respectively, from D. pteronyssinus and/or D. farinae species. Furthermore, 20-47% of patients also have IgEs to allergens from groups 4, 5, 7, 13, 15, 21, and 23. All patients have IgEs to allergens present in mite bodies and feces. MS-based analyses confirmed the presence of mite allergens recorded by IUIS in D. pteronyssinus and D. farinae extracts, with groups 2, 8, 10, 11, 14, and 20 prominent in bodies and groups 1, 6, 18, and 23 well represented in feces. CONCLUSIONS: Mite-specific AIT should rely upon a mixture of D. pteronyssinus and D. farinae extracts, manufactured from both feces and bodies. Such a combination is appropriate to treat children and adult Dermatophagoides-allergic patients from Asia, Europe, and North America.

Allergic airway diseases in a tropical urban environment are driven by dominant mono-specific sensitization against house dust mites.

BACKGROUND: Southeast Asian populations are increasingly affected by allergic airway diseases. Etiology and specific causes, however, are still unknown. The aim of this study is therefore to identify allergens and risk factors for the high prevalence of allergic airway disease in the tropical urban environment. METHODS: Symptoms of allergic rhinitis (AR), asthma, and allergic dermatitis were recorded in two independent cohorts of 576 and 7373 ethnic Chinese individuals living in Singapore. Reactivity against common allergens was determined by skin prick tests (SPT); specific immunoglobulin E (sIgE) titers against 12 common allergens, as well as total serum IgE (tIgE), were measured in the smaller cohort. RESULTS: Immunoglobulin E sensitization was almost exclusively directed against house dust mite (HDM) allergens. More than 80% of individuals were HDM-sIgE positive. Of these, less than 30% also had sIgE for other allergens, and similarly, few of the HDM-sIgE-negative individuals reacted to other allergens. Titers for HDM-sIgE were 8-30 times higher than other non-HDM allergen titers and correlated directly with total serum tIgE levels. Migrants from nontropical countries typically arrived with low or undetectable HDM-sIgE but developed substantial titers in a time-dependent fashion. Importantly, prolonged stay in Singapore also resulted in the manifestation of AR and asthma symptoms, contributing to some of the highest national prevalence rates worldwide. CONCLUSION: In a tropical urban environment, the allergic response is dominated by a single allergen class. The mono-specific IgE sensitization against HDM translates into increased prevalence of allergic airway diseases, which now impact a large proportion of the population in Singapore.

Multiple wheat flour allergens and cross-reactive carbohydrate determinants bind IgE in baker's asthma.

BACKGROUND: Several wheat flour allergens relevant to baker's asthma have been identified in the last 25 years. The aim of this study was to determine the frequency of sensitization to these allergens in German bakers. METHODS: Using recombinant DNA technology, the following wheat flour allergens were cloned, expressed in Escherichia coli and purified: five subunits of the wheat α-amylase inhibitors (WTAI-CM1, WTAI-CM2, WTAI-CM3, WDAI-0.19 and WMAI-0.28), thioredoxin, thiol reductase or 1-cys-peroxiredoxin homologues, triosephosphate-isomerase, αß-gliadin, serpin, glyceraldehyde-3-phosphate-dehydrogenase, a nonspecific lipid transfer protein (nsLTP), dehydrin, profilin and peroxidase. In addition, ImmunoCAPs with the recombinant allergen ω-5-gliadin and two cross-reactive carbohydrate determinants (CCDs), horse radish peroxidase (HRP) and the N-glycan of bromelain (MUXF), were used. Specific IgE was measured in wheat flour-positive sera from 40 German bakers with work-related asthma/rhinitis and 10 controls with pollinosis. RESULTS: Thirty bakers (75%) had IgE to at least one of the 19 single allergens. Most frequent was IgE to WDAI-0.19, HRP and MUXF (25% each), followed by WTAI-CM1 (20%), thiol reductase (16%), WTAI-CM3 (15%), WTAI-CM2 and thioredoxin (12.5%), WMAI-28, triosephosphate-isomerase, αß-gliadin (10%), 1-cys-peroxiredoxin (7.5%), dehydrin, serpin, glyceraldehyde-3-phosphate-dehydrogenase (5%), ω-5-gliadin, nsLTP and profilin (2.5%). Fifteen bakers (38%) had IgE to any α-amylase inhibitor and 12 (30%) to at least one CCD. The controls reacted exclusively to CCDs (80%), profilin (60%), thioredoxin (30%), triosephosphate isomerase and nsLTP (10%). CONCLUSIONS: The single allergen sensitization profiles obtained with 17 recombinant wheat flour allergens and two CCDs revealed no major allergen for German bakers. The highest frequencies were found for α-amylase inhibitors and CCDs.

Collembola are unlikely to cause human dermatitis.

Abstract There have been several unconfirmed case reports of dermatitis caused by Collembola (springtails). We recently investigated two nurses with dermatitis suspected to be caused by Drepanura Schött (Collembola: Entomobryidae). IgE antibodies to Collembola proteins were not detected in sera from the nurses and skin tests with the Collembola extract and crushed whole Collembola were negative in both the nurses and volunteers. This study suggests that the springtail Drepanura may not cause human dermatitis and that other organisms and organic matter that are also found in the moist environment inhabited by Collembola might instead be responsible.

Mite sensitization among Latina women in New York, where dust-mite allergen levels are typically low.

UNLABELLED: In New York (NY), Latinos often have greater asthma morbidity than other ethnicities, and dust-mite sensitization is common despite low allergen levels. We investigated mite allergen exposure and sensitization in atopic and/or asthmatic women, the majority being Puerto Rican. Women (n = 274) recruited for a birth cohort study were visited postnatally. Dust from their homes was analyzed for mite allergens (Der f 1, Der p 1, and Blo t 5). Serum was analyzed for total and allergen-specific IgE. Thirty-seven percent were sensitized to Dermatophagoides pteronyssinus, 34% to Dermatophagoides farinae, and 21% to Blomia tropicalis. Only 5% of NY homes had levels of Der f 1 >2 microg/g; none had Blo t 5 or Der p 1 above this level. Caribbean or Latin American birthplace (a proxy for childhood exposure) was not associated with mite sensitization. Sensitization to D. pteronyssinus and D. farinae was associated with a report of doctor-diagnosed asthma [Odds ratio (OR) = 3.27, P = 0.003; OR = 2.81, P = 0.010, respectively]; sensitization to any mite was associated with asthma medication use in the past 12 months (OR = 3.12, P = 0.004). These associations held even after adjustment for cockroach, mouse, and cat sensitization. PRACTICAL IMPLICATIONS: Despite the low concentrations of mite allergen in our community, many of the women in the atopically enriched cohort were sensitized to mites, even Blomia tropicalis which is typically found only in tropical environments.

Identification and characterization of microsatellite loci in Intsia palembanica (Leguminosae), a valuable tropical timber species.

We present 25 polymorphic microsatellite loci for Intsia palembanica, a highly valued timber species in the Indo-Malayan region, Australia and western Pacific islands. Microsatellite loci were tested for polymorphism across a total of 76 individuals from three natural populations from Irian Jaya, Indonesia. The average number of alleles for these microsatellites was 12.1 per locus, ranging from four to 19. The observed heterozygosity within the natural populations ranged from 0.01 to 0.96. The markers will enable us to evaluate the spatial-temporal population genetic structure and gene flow dynamics among populations. They can also be used for tracking and tracing wood from legally logged concessions.

Home exposures to environmental tobacco smoke and allergic symptoms among young children in Singapore.

BACKGROUND: Research relating environmental tobacco smoke (ETS) exposures have focused on childhood asthma. There have been fewer studies with conflicting results performed on associations of ETS exposures with allergic symptoms. We are interested to see if ETS exposures in the homes are associated with allergic symptoms among preschool children in Singapore where public smoking is banned. METHODS: A cross-sectional study adopting an expanded and modified ISAAC (International Study on Asthma and Allergies in Childhood) questionnaire for the evaluation of asthma and allergies was conducted on 6,794 children attending 120 randomly selected child care centers. Specific information on demographics and ETS exposures was obtained. Adjusted prevalence ratios (PRs) and 95% confidence intervals (CIs) were determined using Poisson multivariate regression with a log-link function and robust variance estimates as recommended for cross-sectional studies. RESULTS: The response proportion was 70.0%, and 4,759 children from 97 centers participated. After adjusting for covariates, it was found that home ETS exposure was associated with increased risks of current symptoms of rhinitis (PR 1.23; 95% CI 1.01-1.50) and rhinoconjunctivitis (PR 1.79; 95% CI 1.26-2.54). These associations followed dose-response trends with respect to number of cigarettes smoked or smokers in the homes. Home ETS exposures were also associated with higher PRs of wheeze, nocturnal cough and doctor-diagnosed asthma. Compared with paternal smoking, higher risks of the above outcomes were found for maternal smoking. CONCLUSION: Home ETS exposure is a risk factor associated with rhinitis and asthma among preschool children.

The effect of ventilation strategies of child care centers on indoor air quality and respiratory health of children in Singapore.

UNLABELLED: This paper reports the effects of ventilation strategies on indoor air quality (IAQ) and respiratory health of children within 104 child care centers (CCCs) in a hot and humid climate. The CCCs were categorized by ventilation strategies: natural (NV), air-conditioned and mechanically ventilated (ACMV), air-conditioned using split units (AC), and hybrid (NV and AC operated intermittently). The concentration levels of IAQ parameters in NV CCCs are characterized by the influence of the outdoors and good dilution of indoor pollutants. The lower ventilation rates in air-conditioned CCCs result in higher concentrations of occupant-related pollutants but lower outdoor pollutant ingress. This study also revealed lower prevalence for most asthma and allergy, and respiratory symptoms in children attending NV CCCs. In multivariate analyses controlled for the effects of confounders, the risk of current rhinitis among children is significantly higher if they attend mechanically ventilated CCCs compared to NV CCCs. Air-conditioned CCCs were also associated with higher adjusted prevalence ratio of severe phlegm and cough symptoms and lower respiratory illness. Finally, children attending CCCs with hybrid ventilation are at high risk for almost all the respiratory symptoms studied. PRACTICAL IMPLICATIONS: This large field study indicates that different ventilation strategies employed by child care centers can cause significant variations in the indoor air quality and prevalence of asthma, allergies and respiratory symptoms of attending children. The higher prevalence rates of allergic and respiratory symptoms among young children, whose immune system is still under-developed, in child care centers, whether fully or partially air-conditioned, suggest that ventilation and plausible growth and propagation mechanisms of allergens and infectious agents be further investigated.

Viability and adipogenic potential of human adipose tissue processed cell population obtained from pump-assisted and syringe-assisted liposuction.

BACKGROUND: Correcting soft tissue defects by autologous fat grafting has been a routine procedure in plastic surgery. Liposuction material for autologous fat grafting is normally obtained using a hand-held syringe. The pump-assisted liposuction technique is not used because of the belief that cells in the harvested fat tissue are necrotic from the physical forces created by the vacuum pump liposuction machine. OBJECTIVE: The metabolism and adipogenic potential of cultured mesenchymal precursor cells processed from pump and syringe lipoaspirates were studied. METHODS: Metabolic estimates were determined with alamarBlue assay. These cells were also induced along the adipogenic lineage with adipogenic induction factors. The extent of adipogenic differentiation was measured using a digital photo counting software. RESULTS: The cells obtained by pump-assisted liposuction are viable, have comparable metabolic activity and adipogenic potential which are comparable to cells using the syringe for aspiration. CONCLUSION: The implications of this study are that pump-assisted liposuction might be a possible alternative to aspirating adipose tissue for reimplantation during reconstruction procedures.

Proteomic analysis of human tears: defensin expression after ocular surface surgery.

Human tear protein profiles were monitored by surface enhanced laser desorption/ionization-time-of-flight mass spectrometry ProteinChip technology (SELDI-TOF ProteinChip) and liquid chromatography-mass spectrometry (LC-MS). Tears were collected from 21 patients scheduled for surgery to remove an ocular surface neoplasm prior to surgery (day 0) and on days 1, 3, and 30 postoperatively. Using this proteomic approach, we verified that three human alpha-defensins (HNP-1, HNP-2, and HNP-3) were significantly up-regulated in their expression after surgery and that their levels decreased to approximately normal by day 30 by which time healing was complete. Further confirmation of the identity of the alpha-defensins in human tears was made by LC purification, trypsin digestion, and ESI-MS/MS analysis of their tryptic digests. The concentrations of HNP-1 and HNP-2 were determined and shown to be markedly increased after ocular surface surgery. The results of the study suggest that human alpha-defensins HNP-1, HNP-2, and HNP-3 are up-regulated after surgery, and may in addition to their antimicrobial properties have an important role in wound healing.

Economic burden of varicella in Singapore--a cost benefit estimate of implementation of a routine varicella vaccination.

Varicella is a common childhood illness that can result in significant morbidity and mortality. As early as 1995, recommendations for routine varicella vaccination have been published, but have not been universally implemented, with cost of vaccination as a major reason. Though available from 1996, the vaccine has yet to be routinely implemented in Singapore. We set out to assess the economic burden of varicella and the cost-benefit of adding a varicella vaccine to the existing immunization schedule in Singapore. In this study, using data from 1994--1995 the direct cost estimates were based on all levels of medical care; inpatient care, emergency room visits, primary health care and medication. Indirect costs were estimated from the cost of time lost by patients and their families attending to medical needs, as well as loss of productivity due to absenteeism. The cost of a vaccination program targeted at 15-month old infants receiving concomitant measles-mumps-rubella immunization was also assessed. The cost-benefit ratio was then estimated. The total cost of varicella in Singapore was estimated to be US$11.8 million per annum. The loss of productivity accounted for a large proportion of the total cost as a lot of parents took leave when their children were ill. The estimates of total cost represent approximately US$188 per varicella case per year. In comparison, the cost of a vaccination program was found to be US$3.3 million per annum. The cost per case averted was US$104. From a societal point of view, for every dollar invested in a vaccination program, we would save about US$2 dollars.

Laboratory assessment of the efficiency of encasing materials against house dust mites and their allergens.

BACKGROUND: The current recommendation to reduce mite allergen exposure for mite-sensitive individuals is to use allergen-impermeable bed coverings. As these covers are made of various kinds of materials, they vary in quality. The objective of this study was to investigate the efficiency of different covering materials against house dust mites and their allergens in vitro. METHODS: Four types of materials including (1) plastic cover, (2) polyurethane-coated cover, (3) non-woven covers, (4) tightly woven microfiber covers and a regular cotton bed sheet (as a control) were evaluated using three methods: (i) heat escape method, (ii) Siriraj chamber method and stereomicroscopy, scanning electron microscopy and (iii) enzyme-linked immunosorbent assay (ELISA). RESULTS: We found that there was a statistically significant difference in allergen permeability among four types of coverings (P < 0.001). In terms of the impermeability to mites and their allergens, plastic- and polyurethane-coated covers were observed to be the best, followed by non-woven, woven and cotton-based bed sheets. A regular cotton-based bed sheet allows a significant amount of leakage of mite allergens. Both woven and non-woven material are efficient barriers against mite allergen in terms of impermeability. However, with regard to mite colonization, non-woven covers have the drawback of mites being able to penetrate and colonize within the fabric fibers. Woven covers are therefore recommended because of their major advantages of not allowing the colonization of mites within the fabric, being easy to clean, and comfortable. CONCLUSION: The three assessment methods used in this study could be useful as a primary approach to evaluate the quality of covering materials in vitro using both pore size and ability to be colonized by mites on the materials as the key factors.

The upper and lower airway responses to nasal challenge with house-dust mite Blomia tropicalis.

BACKGROUND: The house dust mite Blomia tropicalis (B. tropicalis) was found to be the most prevalent domestic mite in Singapore. However, its pathogenicity in allergic airway diseases remains to be investigated. METHODS: Twenty adults with persistent allergic rhinitis (PAR) were studied. Five had a history of asthma, and all were asymptomatic except one who was under treatment with low-dose inhaled corticosteroid. Nasal challenge was carried out by nasal spray with phosphate-buffered saline (PBS) and with increasing concentrations of crude B. tropicalis extracts (0.6, 6.0 and 60 micro g/ml) at 15 min intervals. Subjective symptom scores and absolute number of sneezes were recorded together with objective measurements of spirometry (forced expiratory volume in 1 s, FEV1) and acoustic rhinomanometry (volume of the nasal cavity). These were performed at baseline, 5 min after each incremental challenge, and 30 min, 1 h, 3 h, 5 h and 7 h after the last challenge. Meanwhile, concentrations of mediators in nasal secretions (tryptase, leukotriene C4 (LTC4) and eosinophil cationic protein (ECP)) were measured in nasal aspirate samples at similar time intervals. An identical (control) challenge procedure with PBS alone was repeated in seven patients after a washout period of at least 2 weeks. RESULTS: Significant increases in the subjective and objective nasal symptoms, together with a significant increase of tryptase and LTC4 concentrations in nasal secretion, were found 5 min after each challenge with B. tropicalis, but not with PBS. There was no definitive pattern of the late-phase nasal response in either subjective symptoms or objective measurements. Three patients (3/5) with a history of asthma showed a fall in FEV1 readings (33%, 22% and 11% from baseline, respectively) at 7 h post challenge with concomitant mild wheezing in the night. CONCLUSIONS: Our study demonstrates direct evidence of allergic nasal response to B. tropicalis in sensitized adults. It shows that nasal provocation may also provoke concomitant asthmatic symptoms during the late-phase reaction, especially in people with a history of asthma.

Seasonal variation in respiratory syncytial virus chest infection in the tropics.

Respiratory syncytial virus (RSV) is the most important cause of lower respiratory tract infection (LRTI) in young children. We determined if there was a seasonal variation in Malaysia in the incidence of RSV infection in young children admitted with LRTI, and possible associations of RSV infection with local meteorological parameters. A total of 5,691 children, aged less than 24 months and hospitalized with LRTI (i.e., bronchiolitis and pneumonia) between 1982-1997, were included in this study. Nasopharyngeal samples were collected and examined for RSV by immunofluorescence, viral culture, or both. Seasonal variations were determined by analyzing the monthly RSV-positive isolation rate via time series analysis. Possible correlations with local meteorological parameters were also evaluated.RSV was isolated in 1,047 (18.4%) children. Seasonal variations in RSV infection rate were evident and peaked during the months of November, December, and January (test statistics [T] = 53.7, P < 0.001). This seasonal variation was evident for both bronchiolitis and pneumonia categories (T = 42.8 and 56.9, respectively, P < 0.001). The rate of RSV infection appeared to correlate with the monthly number of rain days (r = 0.26, P < 0.01), and inversely with the monthly mean temperature (r = -0.38, P < 0.001). In the tropics, seasonal variations in the incidence of RSV infection are evident, with an annual peak in November, December, and January. This information provides a guide for healthcare provisions and implementation of RSV prevention.

Genetic susceptibility to asthma and atopy among Chinese in Singapore--linkage to markers on chromosome 5q31-33.

BACKGROUND: Asthma and atopy are complex genetic traits, influenced by the interaction of multiple genes and environmental factors. Linkage of these traits to chromosome 5q31-33 has been shown in other populations, but has not been well studied in the Chinese. We studied linkage between asthma and atopy with markers on chromosome 5q31-33 in the Singapore Chinese. This region contains many candidate genes, including the cytokine gene cluster. METHODS: We recruited 88 Chinese families with at least two affected offspring, totaling 373 subjects, with 125 and 119 sib-pairs for atopy and asthma, respectively. All individuals were genotyped with 19 polymorphic microsatellite markers spanning a distance of 41 cM along chromosome 5q31-33. Affected sib-pair and multipoint linkage analysis was performed. RESULTS: There was evidence for linkage of the asthma and atopy phenotypes with three markers, D5S2110, D5S2011, and D5S412 (P values of 0.001 to 0.00001). Multipoint analysis further substantiated this (nonparametric linkage scores of 1.8-2.9). These findings suggest that susceptibility genes for asthma and atopy are found in this region in the Chinese. CONCLUSION: This study has shown linkage of atopy and asthma to chromosome 5q31-33 in a heterogeneous Chinese population. These findings further substantiate the notion that chromosome 5q31-33 contains "universally" important susceptibility genes for these traits.

Immunochemical characterization of edible bird's nest allergens.

BACKGROUND: We have previously described anaphylaxis induced by edible bird's nest (BN) and demonstrated that this condition is IgE mediated. OBJECTIVES: This study aimed at describing the immunochemical properties of the BN allergens. Comparative studies between 3 commercially available sources (according to the country of origin) of BN were also made. METHODS: Crude extracts of commercially available processed BN from Sarawak (Malaysia), Thailand, and Indonesia and fresh unprocessed BN from the caves of Sarawak were obtained by means of aqueous extraction. Specific IgE toward these sources were determined by using fluorescence allergosorbent tests (FASTs). Cross-reactivity studies between the 3 sources of commercially available processed BN were carried out by means of FAST inhibition. Immunochemical characterization by means of IgE immunoblot, periodate treatment, and heat stability studies were carried out on fresh unprocessed BN from Sarawak. RESULTS: Serum from allergic patients showed differences in IgE binding to the 3 sources of commercially available BN, with the highest levels of specific IgE recorded with the Sarawak source (P <.0001). Of these, only the Sarawak and Thailand sources showed considerable cross-reactivity. Further work on the unprocessed fresh Sarawak source identified a putative 66-kd major allergen containing several isoforms. Periodate treatment resulted in loss of IgE binding. Despite a progressive decline in the molecular weights of allergens on SDS-PAGE with increasing periods of boiling, IgE binding, as assessed by means of FAST, was not affected. N-terminal sequence of the major putative allergen (66 kd) showed homology to a domain of an ovoinhibitor precursor in chicken (SWISS-PROT accession No. P10184). CONCLUSIONS: We have described the immunochemical properties of BN allergens. Edible BN from different sources are allergenically dissimilar. The putative major allergen is a 66-kd protein.

Edible "bird's nest"-induced anaphylaxis: An under-recognized entity?

The Chinese delicacy "bird's nest" is the most common cause of food-induced anaphylaxis requiring hospitalization among Chinese children in Singapore. This investigation has established an immunoglobulin E-mediated cause and has characterized the major putative allergens.