Resistance of Commercial Bed Covers against Faecal Pellets of House Dust Mites:Behaviour of Seams and Zippers

This study evaluated the allergen impermeability against airborne allergens of dust mite droppings through all parts of commercial bed covers, including surface seams and zippers. Specimens were taken from places with and without seams and zipper. A novel penetration cell was developed to expose the specimens to an inoculum of purified mite droppings that was assessed for its allergen content Der p1 prior to the penetration tests. Using covers of different construction and material, the penetration level increased significantly in the presence of seams and zippers and could reach up to 6% depending on the seam’s/zipper’s characteristics and quality. Therefore, zippers and seams have to be considered as access points for the penetration of mite droppings. As for the penetration of airborne mite particles through the zipper, the penetration level was greatly attenuated by the presence of a cover strip. Depending on the respective quality and the construction type, the mite allergen Der p1 penetrated most likely through the zipper and seams of the specimens, already after a single laundry cycle. Hence, laundry may compromise the barrier performance and proves to be an important quality feature. In all samples, the textile surface showed sufficient allergen impermeability. Our conclusions provide recommendations to both manufacturers and users.

HOUSE DUST MITE ALLERGEN (Derp1 AND Blot5) LEVELS IN ASTHMATICS' HOME IN HONGKONG

To measure Derp1 and Blot5 allergen levels in asthmatics’ homes in Hongkong. Methods Seventy houses were enrolled for a mite indoor environment study. Dust samples were obtained from two sites of each patients’ house: bed and floor. Derp1 and Blot5 levels were quantified by a two-site monoclonal antibody-based ELISA technique. Results The levels of Derp1 allergens found in bed (geometric mean (GM) 3.43 μg/g of dust; 95%CI, 1.89-4.96 μg/g) and on the floor (GM 1.12 μg/g of dust; 95%CI, 0.71-1.53 μg/g) indicated significant differences (P = 0.005). However, the levels of Blot5 allergens found in bed (GM 19.00 μg/g of dust; 95%CI, 0.89-38.90 μg/g) and on the floor (GM 6.14 μg/g of dust; 95%CI, 0.40-11.90 μg/g) showed no statistically significant difference. In addition, in regards to the exposure index for Derp1 and Blot5 allergens found in bed and on the floor, 17.6% in bed and 8.6% on the floor had levels of Blot5 ≥ 10 μg/g of dust, higher than those obtained for Derp1 (7.2% and 0% in bed and on the floor respectively, P < 0.05); higher percentages in bed and on the floor (25.0% and 35.7%) were observed for levels of Blot5 = 0 μg/g of dust as compared with Derp1 in bed and on the floor (4.3% and 14.5% respectively, P < 0.05). Conclusions Derp1 and Blot5 are the major allergens found in this regional study, Blot5 is a more potent allergen in Hongkong, probably reflecting the high level of exposure to Blomia tropicalis (Bt). Bt and Dermatophagoides pteronyssinus(Dp) allergens should be included for precise diagnosis and effective immuno-therapeutic treatment of mite allergy in Hong-kong.

Acaricidal activities of some essential oils and their monoterpenoidal constituents against house dust mite,Dermatophagoides pteronyssinus (Acari:Pyroglyphidae)

The acaricidal activities of fourteen essential oils and fourteen of their major monoterpenoids were tested against house dust mites Dermatophagoides pteronyssinus. Five concentrations were used over two different time intervals 24 and 48 h under laboratory conditions. In general, it was noticed that the acaricidal effect based on LC50 of either essential oils or monoterpenoids against the mite was time dependant. The LCso values were decreased by increasing of exposure time. Clove, matrecary, chenopodium, rosemary, eucalyptus and caraway oils were shown to have high activity. As for the monoterpenoids, cinnamaldehyde and chlorothymol were found to be the most effective followed by citronellol. This study suggests the use of the essential oils and their major constituents as ecofriendly biodegradable agents for the control of house dust mite, D. pteronyssinus.

House dust mite allergen Der f enhanced bronchial epithelial cell cytokine expression

The house dust mites (Dermatophagoides farinae, Derf) are the major source of aeroaller gens implicated in the expression of atopic disorders, including asthma, allergic rhinitis and atopic dermatitis. In particular, strong circumstantial evidence suggests that house dust mite antigens are important precipitating factors of asthma. Many house dust mite allergens are proteases that can elicit airway inflammation by stimulating the release of cytokines from bronchial epithelial cells. To investigate whether Der f allergen proteases induced cytokine production from the epithelial cell line BEAS-2B, BEAS-2B cells were cultured with 4 different concentrations of Derf (0.02, 0.2, 2, 20μg/ml) for 24-96 h, after which supernatants were assayed for interleukin (IL)-6 and IL-8 with ELISA. Reverse transcription-PCR was also performed. The cell sheets were intact throughout the observation in control group without any exposure to Der f antigen. In the experimental groups cells treated with Der f allergen showed changes in the anchorage status of the monolayer. There was a significant increase in the level of cytokine production compared with the untreated sample. The release of IL-6 and IL-8 increased in a concentration-dependent manner (P <0.05, respectively) with the addition of increasing dosage of Der f to the cell sheets. Levels of IL-6 and IL-8 began to rise at 24 h and 48 h after allergen exposure, and they increased significantly in the supematants at 72 h and 96 h. At the same time the concentration dependence of induction of IL-6 and IL-8 expression as well as an increase in the expression of IL-6 and IL-8 mRNA manifested evidently. HDM-induced airway inflammation may include Der f-mediated release of inflammatory mediators, and the proteolytic activity of an allergen may stimulate the release of proinflammatory cytokines from human bronchial epithelium. It is suggested that IL-6 and IL-8 production by bronchial epithelial cells may play a role in the pathogenesis of allergic asthma.

Indoor Allergen Levels and Household Distributions in Nine Cities Across China

Objective Chinese allergic subjects have high levels of sensitization to house dust mite （HDM） and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households. Methods Dust samples were collected from nine cities. Major allergens Der p 1 and Der f I from Dermatophagoides pteronyssinus and D. farinae, and specific antigens of Blomia tropicalis, Tyrophagus putrescentiae, Acarus siro, and cockroach species Blattella germanica and Periplaneta americana were measured by ELISA.Results HDM allergens were found in dust samples from bedding in 95% of the Chinese households. The median levels varied from 〈0.006 to 9.2 μg/g of dust, depending on the city. The percentages of households having HDM allergen levels associated with the risk of developing allergy sensitization and asthma were 65% and 25%, respectively. Specific antigens of the storage mite and cockroach were only found in samples from the southern and tropical regions of China. Levels of mite allergens were generally higher in samples from bedding compared to samples from the living room, even for storage mites, whereas levels of cockroach antigens were higher in the living room samples.Conclusion HDM allergens are present in bedding dust samples from most Chinese households. Cities in southern and central China have relatively high levels of HDM major allergens compared to cities in northern and western China. Antigens of storage mites and cockroaches are not as common as HDM allergens.

FGF2 is overexpressed in asthma and promotes airway in airway epithelial cells

Background: Airway inflammation is the core pathological process of asthma, with the key inflammatory regulators incompletely defined. Recently, fibroblast growth factor 2(FGF2) has been reported to be an inflammatory regulator;however, its role in asthma remains elusive. This study aimed to investigate the immunomodulatory role of FGF2 in asthma.Methods: First, FGF2 expression was characterised in clinical asthma samples and the house dust mite(HDM)-induced mouse chronic asthma model. Second, recombinant mouse FGF2(rm-FGF2) protein was intranasally delivered to determine the effect of FGF2 on airway inflammatory cell infiltration. Third, human airway epithelium-derived A549 cells were stimulated with either HDM or recombinant human interleukin-1β(IL-1β) protein combined with or without recombinant human FGF2. IL-1β-induced IL-6 or IL-8 release levels were determined using enzyme-linked immunosorbent assay, and the involved signalling transduction was explored via Western blotting.Results: Compared with the control groups, the FGF2 protein levels were significantly upregulated in the bronchial epithelium and alveolar areas of clinical asthma samples [(6.70±1.79) vs.(16.32±2.40), P=0.0184;(11.20±2.11) vs.(21.00±3.00), P=0.033, respectively] and HDM-induced asthmatic mouse lung lysates [(1.00±0.15) vs.(5.14±0.42),P<0.001]. Moreover, FGF2 protein abundance was positively correlated with serum total and anti-HDM IgE levels in the HDM-induced chronic asthma model(R^(2)=0.857 and 0.783, P=0.0008 and 0.0043, respectively). Elevated FGF2protein was mainly expressed in asthmatic bronchial epithelium and alveolar areas and partly co-localised with infiltrated inflammatory cell populations in HDM-induced asthmatic mice. More importantly, intranasal instillation of rm-FGF2 aggravated airway inflammatory cell infiltration [(2.45±0.09) vs.(2.88±0.14), P=0.0288] and recruited more subepithelial neutrophils after HDM challenge [(110.20±29.43) cells/mm^(2) vs.(238.10±42.77) cells/mm^(2), P=0.0392]without affecting serum IgE levels and Th2 cytokine transcription. In A549 cells, FGF2 was upregulated through HDM stimulation and promoted IL-1β-induced IL-6 or IL-8 release levels [up to(1.41±0.12)-or(1.44±0.14)-fold change vs.IL-1β alone groups, P=0.001 or 0.0344, respectively]. The pro-inflammatory effect of FGF2 is likely mediated through the fibroblast growth factor receptor(FGFR)/mitogen-activated protein kinase(MAPK)/nuclear factor kappa B(NF-κB)pathway.Conclusions: Our findings suggest that FGF2 is a potential inflammatory modulator in asthma, which can be induced by HDM and acts through the FGFR/MAPK/NF-κB pathway in the airway epithelial cells.

Effect of Sublingual Immunotherapy on Level of Cytokines in PBMCs of Patients with Allergic Asthma

This study examined the possible mechanism of sublingual immunotherapy（SLIT） in the treatment of allergic asthma.Forty asthma patients allergic to dust mite were enrolled.They received SLIT with dermatophagoides farinae（Der.f） drops for one year.Thirty healthy subjects served as controls.The levels of IL-4 and IFN-γ of peripheral blood mononuclear cells（PBMCs） were determined in allergic asthma patients before and after the SLIT as well as the healthy subjects.The results showed that the level of IL-4 was substantially increased and that of IFN-γ remarkably decreased in the patients before the SLIT as compared with those in the healthy subjects（P0.05）.After the SLIT,the level of IL-4 was significantly reduced and that of IFN-γ elevated in these allergic asthma patients.It was concluded that sublingual immunotherapy is effective for patients with allergic asthma.And it may work by regulating the balance of Th1/Th2 through changing the expression of IL-4 and IFN-γ in PBMCs.

Comparison of Three Methods of Protein Extraction from Dermatophagoides Pteronyssinus for Two-dimensional Electrophoresis

Obiective To explore an effective method of Dermatophagoides pteronyssinus protein extraction suitable for two-dimensional electrophoresis （2-DE） analysis. Methods The extracts of Dermatophagoides pteronyssinus were prepared with Coca＇s solution, lysis buffer of 2-DE, and Trizol reagent, respectively. Bicinchoninic acid （BCA） assay was used to determine the total protein concentration of the samples. The efficiency of different protein extraction methods were evaluated with 2-DE analysis. Results The concentrations of extracted protein by methods of Coca＇s solution, lysis buffer, and Trizol reagent were 0.63 g/L, 0.90 g/L, and 0.80 g/L, respectively. The 2-DE analysis results showed that some protein spots in low molecular weight （LMW） range could be detected with the Coca＇s solution method. With the lysis buffer of 2-DE method, more protein spots in LMW range could be detected, while the medium molecular weight （MMW） protein spots were absent. Several MMW protein spots （174-178 kD and 133 kD） and more LMW protein spots were detected withTrizol reagent method. Conclusions Among Coca＇s solution, lysis buffer of 2-DE, and Trizol reagent, the concentration of extracted protein of Dermatophagoides pteronyssinus by lysis buffer of 2-DE is the highest. However, most protein components of Dermatophagoides pteronyssinus purified mite bodies can be extracted by Trizol reagent, which may generally reflect the whole profile of Dermatophagoides pteronyssinus allergens.

House dust mite allergens and nitrated products: Identification and risk assessment in indoor dust

Air pollutants can potentially lead to nitration of allergic proteins,thus promoting sensitization of these allergens.However,little is currently known about the nitration status of house dust mite(HDM)allergens.We identified the occurrence of nitrated products of two major HDM allergens Der f 1 and Der p 1 in dust samples collected from college dormitories in eastern China and assessed their associated health risk.The results showed that both non-nitrated and nitrated forms of the two allergens were detected in the dust in the range of non-detected(ND)-10.6,1.44-15.4,ND-22.4,ND-7.28μg/g for non-nitrated Der f 1,nitrated Der f 1,non-nitrated Der p 1 and nitrated Der p 1,respectively.The median rates of nitration were determined as 74.0%for Der f 1 and 20.4% for Der p 1 at consideration of one nitration site.Further analysis reveals that the levels of HDM allergens and their nitrated products were found to be generally higher during winter,in dormitories of lower altitude and with female occupants.Furthermore,the calculated risk indexes were at considerably high levels.Our findings suggest that nitrated HDM allergens have already accumulated in the environment at such significant levels and their associated health risk calls for our immediate attention.

Do the microorganisms from laboratory culture spent growth medium affect house dust mite fitness and microbiome composition?

The interaction of house dust mites(HDM)and microorganisms is the key factor in the survival of these mites in human-made environments.Spent growth medium(SPGM)provides the rest of the dict,along with dead mite bodies and microorganisms.SPGM represents a source of microorganisms for the recolonization of mite food and the mite digestive tract.An experiment was performed to observe how adding SPGM to the HDM diet affects HDM population growth,the microbiome composition and the microbial respiration in microcosms.We analyzed American house dust mite(Dermatophagoides farinae)and European house dust mite(Dermatophagoides pleronyssinus)originating from control diets and diets treated with an extract of SPGM from 1-and 3-month-old mite cultures.The microbiome was described using 16S and 18S barcode sequencing.The composition of the bacterial and fiungal microbiomes differed between the HDM species,but the SPGM treatment influenced only the bacterial profile of D.farinae.In the D.farinae microbiome of specimens on SPGM-treated dicts compared to those of the control situation,the Lactobacillus profile decreased,while the Candinium,Staphylococ-cus,Acinetobacter,and Sphingomonas profiles increased.The addition of SPGM extract decreased the microbial respiration in the microcosms with and without mites in almost all cascs.Adding SPGM did not influence the population growth of D.farinae,but it had a variable effect on D.pteronyssimus.The results indicated that the HDM are marginally influenced by the microorganisms in their feces.

Effect of house dust mite immunotherapy on interleukin-10-secreting regulatory T cells in asthmatic children

Background Subcutaneous specific immunotherapy has been demonstrated to be capable of inducing T-cell regulatory response.Interleukin-10 （IL-10） plays a crucial role in inducing allergen-specific tolerance.However the reports of the changes of IL-10 in house dust mite （HDM）-specific immunotherapy were varied.The aim of this study was to evaluate the function of IL-10-secreting regulatory T cells in asthma children successfully treated with HDM immunotherapy.Methods Peripheral blood mononuclear cells （PBMCs） were isolated from 27 patients following 1.5--2 years of HDM-specific immunotherapy （SIT, SIT group） and from 27 matched treated asthmatic children allergic to HDM （asthmagroup）.After 48 hours of in vitro stimulation with HDM extracts, IL-10-secreting regulatory T cells were measured by four colour flow cytometry.Sera were tested for allergen-specific IgG4 and IgE using the Immuno CAP 100 assay.Results PBMCs from children undergoing immunotherapy following HDM extracts stimuli produced significantly more IL-10 compared with the asthma group.The frequency of iTreg cells and aTreg cells increased in SIT group after HDM stimulation, while it was not affected in the asthma group.Among the iTreg cells and aTreg cells, the frequency of CD4＋CD25-Foxp3-IL-10＋ Treg cells increased the most which was 2 times higher than that in unstimulated cultures in SIT group.The levels of HDM-specific IgG4 of SIT group was significiently higher compared with asthma group, but there was no correlation of the levels of HDM-specific IgG4 and IL-10 secreting Treg cells.Conclusions HDM-specific immunotherapy can successfully upregulate the frequency of IL-10-secreting Treg cells.CD4＋CD25-Foxp3-IL-10＋ Treg cells may play a key role in inducing the immune tolerance in HDM-specific immunotherapy.

Caspase-1 activation by NLRP3 inflammasome dampens IL-33-dependent house dust mite-induced allergic lung inflammation

The cysteine protease caspase-1(Casp-1)contributes to innate immunity through the assembly of NLRP3,NLRC4,AIM2,and NLRP6 inflammasomes.Here we ask whether caspase-1 activation plays a regulatory role in house dust mite(HDM)-induced experimental allergic airway inflammation.We report enhanced airway inflammation in caspase-1-deficient mice exposed toHDMwith a marked eosinophil recruitment,increased expression of IL-4,IL-5,IL-13,aswell as full-length and bioactive IL-33.Furthermore,mice deficient for NLRP3 failed to control eosinophil influx in the airways and displayed augmented Th2 cytokine and chemokine levels,suggesting that the NLPR3 inflammasome complex controls HDM-induced inflammation.IL-33 neutralization by administration of soluble ST2 receptor inhibited the enhanced allergic inflammation,while administration of recombinant IL-33 during challenge phase enhanced allergic inflammation in caspase-1-deficient mice.Therefore,we show that caspase-1,NLRP3,and ASC,but not NLRC4,contribute to the upregulation of allergic lung inflammation.Moreover,we cannot exclude an effect of caspase-11,because caspase-1-deficient mice are deficient for both caspases.Mechanistically,absence of caspase-1 is associated with increased expression of IL-33,uric acid,and spleen tyrosine kinase(Syk)production.This study highlights acritical role of caspase-1 activation andNLPR3/ASCinflammasomecomplex in the down-modulation of IL-33 in vivo and in vitro,thereby regulating Th2 response in HDM-induced allergic lung inflammation.

Dust mite allergen-specific immunotherapy increases IL4 DNA methylation and induces Der p-specific T cell tolerance in children with allergic asthma

Allergen-specific immunotherapy(allergen-SIT)is a highly effective treatment for children with allergic asthma(AA),an immune-mediated chronic disease leading to bronchial muscle hypertrophy and airway obstruction in response to specific allergens.T helper cells and secreted cytokines play important roles in the pathogenesis of asthma,and epigenetic modulation controls genes important for T cell development and cytokine expression.This study evaluated T helper cell-secreted cytokines and DNA methylation patterns in children treated with Dermatophagoides pteronyssinus(Der p)allergen-SIT.Our results showed that after Der p challenge,peripheral blood mononuclear cells(PBMCs)from the SIT group,compared with the non-SIT AA group,produced lower levels of IL-4,IL-5 and IL-2.The SIT group,compared with the AA group,exhibited decreased sensitivity to the Der p allergen,concurrent with IL-4 down-modulation due to increased promoter DNA methylation,as estimated in PBMCs.Our results showed that SIT decreased IL-4 and IL-5,and inhibited T cell proliferation,by inhibiting IL-2 production after the specific allergen challenge.These results suggest that decreased IL-2 production and increased IL-4 cytokine promoter methylation is a potential mechanism of Der p-specific allergen desensitization immunotherapy.

Prevalence of sensitivity to cockroach allergens and IgE crossreactivity between cockroach and house dust mite allergens in Chinese patients with allergic rhinitis and asthma

Background Cockroaches are an important indoor allergen source causing allergic rhinitis and asthma. The aim of this study was to investigate the cockroach prevalence in mainland of China and the cross-reactivity of IgE between cockroach and house dust mite allergen in Chinese patients.Methods The cockroach sensitization pattern was based on a skin prick test （SPT） obtained from a national multicenter prevalence study, in which 6304 patients from 25 allergy centers across China participated. Factors, including different regions of China, age, gender and the correlations between the American and German cockroaches and house dust mite Der p were investigated. Eighteen out of 1236 clinical sera from south China were selected to perform the cross-inhibition assay between house dust mites and cockroaches.Results Totally 25.7% of patients were SPT positive to the American cockroach （Periplaneta Americana, Per a） and 18.7% SPT positive to the German cockroach （Blattella germanica, Bla g）. The prevalence of positive cockroach SPT was higher in southern than in northern China, higher in adults than in children, and higher in males than in females.Patients had relatively low levels of cockroach SPT reactions, mainly class 1 or 2. Of the SPT positive cockroach patients,88% were also SPT positive to house dust mite （Dermatophagoides pteronyssinus, Der p）. An IgE cross-inhibition study confirmed that Der P sensitization could cause false positive SPT reactions against cockroach.Conclusions A relatively high prevalence of cockroach sensitivity was found in mainland of China. However, a cross-inhibition study showed that only a small number of patients appear to have Bla g and/or Per a as primary sensitizing source. The importance of cockroaches as a risk factor for sensitization and triggers of allergic symptoms in mainland of China needs to be further investigated.

Allergic rhinitis facts from an Irish pediatric population

Objective:Assessing the main allergens in the pediatric population from the largest urban area in the country.Methods:Clinical letters of patients referred with possible allergic rhinitis(AR)were retrospectively reviewed over the past 5 years.Results:Five hundred and fifty‐five patients were included.Males suffer twice as often with AR than females and have high titers of allergens.House dust mites(44.7%)and grass pollen(29%)were the main allergens in our area,with 48%of patients sensitized to both allergens.Half of the patients had the diagnosis of AR confirmed with positive allergen‐specific tests.For the other half,the diagnosis was based on a clinical assessment performed by a pediatric otolaryngologist.Conclusions:Half of suspected AR children have environmental allergen sensitivity confirmed by testing,and a large number had a clinical diagnosis of AR after an otolaryngology consultation.Our findings can help clinicians to initiate AR treatment considering the most problematic allergens in the area.

A fifteen-year review of skin allergy testing in Irish patients with symptomatic rhinitis

Objective:Our objective was to review skin prick allergy testing(SPAT)results in patients with symptomatic rhinitis in an Irish population.Methods:A fifteen-year retrospective review of our database of symptomatic patients with rhinitis was performed.All patients who had SPAT performed during this interval were included.Data was analysed in terms of demographics and dominant allergens.Results:1158 patients were included.617 Females vs 541 Males.Age range five to eighty-five years old.Mean age thirty-four years.49%of our patients tested positive to at least one aeroallergen.The most common allergens were dust mites(23%)and timothy grass(22%).Patients born during the Irish pollen season(April-July)were between 5 and 7 times more likely to be sensitive to timothy and ryegrass pollens compared to others tested.241 patients had both SPAT and serum allergen specific IgE testing(SASIgET)performed;positive results were consistent between both groups.Conclusion:Results demonstrated that half of our patients with symptomatic rhinitis had allergen sensitisation.Dust mites and grass were the main allergens in our area.Our nurse led clinic has allowed efficient patient education and the development of a unique Irish SPAT database.Retesting a patient with a known allergy test result it is not indicated.

Suppression of Immunotherapy on Group 2 Innate Lymphoid Cells in Allergic Rhinitis

Background： Group 2 innate lymphoid cells （ILC2s） are regarded as a novel population of lineage-negative cells that induce innate Type 2 responses by producing the critical Th2-type cytokines interleukin （IL）-5 and IL-13. ILC2s as key players in the development of allergic rhinitis （AR） have been proved, however, the effect of subcutaneous immunotherapy （SCIT） with dermatophagoides pteronyssinus extract （Der p-SCIT） on ILC2s in AR patients is not clear. This study aimed to investigate the response of ILC2s of peripheral blood in house dust mites （HDM）-sensitized Chinese patients with AR who received SCIT with Der P extract. Methods： Seven healthy controls without symptoms of AR who had negative reactions to any of the allergens from skin-prick testing, nine patients diagnosed with persistent AR according to the Allergic Rhinitis and its Impact on Asthma （ARIA） guidelines, and 24 AR patients who received Der p-SCIT for 1.0-3.5 years were recruited for the study. ILC2s in the peripheral blood were evaluated using flow cytometry. The severity of their symptoms of all participants was rated based on the Total 5 symptom score. Results： Among 40 participants, 9 AR patients were assigned to the untreated group, 24 AR patients receiving Der p-SCIT were assigned to the immunotherapy group, and 7 healthy controls without symptoms of AR were assigned to healthy control group. The mean Total 5 symptom score of immunotherapy group was significantly lower than that of untreated group （4.3 ± 1.4 vs. 10.1 ± 2.5, P 〈 0.001 ）. Similarly, the levels of ILC2s in the peripheral blood ofimmunotherapy group were significantly reduced compared with that in untreated group （P 〈 0.001 ）, but were not significantly different from healthy controls （P = 0.775）. Further subgroup analysis based on the duration of SCIT therapy （ 1.0-2.0 years [SCIT1-2], 2.0-3.0 years [SCIT2_3], and 3.0-3.5 years [SCIT3_3.5]） showed that the percentage of ILC2s was not significantly different between SCIT1-2, SCIT2-3, and SCIT3-3.5 groups （SCIT1-2 vs. SCIT2-3： P = 0.268; SCIT1-2, vs. SCIT3-3.5： P = 0.635; and SCIT, 3 vs. SCIT3-3.5： P = 0.787）. Conclusions： The present study highlighted the suppression ofDer p-SCIT on ILC2s in HDM-AR patients. ILC2s identified in peripheral blood can be used as an effective biomarker for Der p-SCIT.

The chemosensory appendage proteome of Amblyomma americanum （Acari： Ixodidae） reveals putative odorant-binding and other chemoreception-related proteins

Proteomic analyses were done on 2 chemosensory appendages of the lone star tick, Amblyomma americanum. Proteins in the fore tarsi, which contain the olfactory Haller＇s organ, and in the palps, that include gustatory sensilla, were compared with proteins in the third tarsi. Also, male and female ticks were compared. Proteins were identified by sequence similarity to known proteins, and by 3-dimensional homology modeling. Proteomic data were also compared with organ-specific transcriptomes from the tick Rhipicephalus microplus. The fore tarsi express a lipocalin not found in the third tarsi or palps. The fore tarsi and palps abundantly express 2 proteins, which are similar to insect odorant-binding proteins （OBPs）. Compared with insect OBPs, the tick OBP-like sequences lacked the cysteine absent in C-minus OBPs, and 1 tick OBP-like sequence had additional cysteines that were similar to C-plus OBPs. Four proteins similar to the antibiotic protein microplusin were found： 2 exclusively expressed in the fore tarsi and 1 exclusively expressed in the palps. These proteins lack the microplusin copper-binding site, but they are modeled to have a significant internal cavity, potentially a ligand-binding site. Proteins similar to the dust mite allergens Der p7 and Der f 7 were found differentially expressed in female fore tarsi. A protein exclusively expressed in the fore tarsi has similarities to Neto, which is known to be involved in clustering ofionotropic glutamate receptors. These results constitute the first report of OBP-like protein sequences in ticks and point to several research avenues on tick chemosensory reception.