Dermatophagoides farinae induces conjunctival epithelial cell damage to promote neutrophil migration and neutrophil extracellular traps formation / 中国血吸虫病防治杂志

Objective To investigate the mechanisms underlying allergic conjunctivitis caused by conjunctival epithelial cell damage, neutrophil migration and neutrophil extracellular traps (NETs) formation induced by crude extracts of Dermatophagoides farinae mite (CDM). Methods Human conjunctival epithelial cells were stimulated with 500, 1 000, 2 000, 4 000 ng/mL, and the expression levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and IL-8 were detected using quantitative real-time PCR (qPCR) assay and enzyme-linked immunosorbent assay (ELISA). The culture supernatant of human conjunctival epithelial cells was collected and co-cultured with neutrophils. Neutrophil migration was measured using Transwell migration assay, and the expression of NETs markers myeloperoxidase (MPO) and citrullinated histone H3 (CitH3) was quantified using immunofluorescence staining. Neutrophils were stimulated with phorbol 12-myristate 13-acetate (PMA), and then NETs were collected for treatment of human conjunctival epithelial cells. Cell apoptosis was detected using flow cytometry, and the levels of IL-6, TNF-α, IFN-γ and IL-8 were measured in the cell culture supernatant using ELISA. Results Treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL up-regulated IL-6, TNF-α, IFN-γ and IL-8 expression in human conjunctival epithelial cells. Following treatment with CDM at concentrations of 2 000 ng/mL and 4 000 ng/mL, the culture supernatant of human conjunctival epithelial cells promoted neutrophil migration and induced increases in the staining intensity of MPO and CitH3. In addition, increased NETs triggered the apoptosis of human conjunctival epithelial cells and IL-6, TNF-α, IFN-γ and IL-8 secretion in the culture supernatant of human conjunctival epithelial cells. Conclusions CDM induces human conjunctival epithelial cell damages, thereby promoting neutrophil migration and NETs formation, while the release of NETs further aggravates human conjunctival epithelial cell damages.

miR-142-3p alleviates house dust mite induced airway inflammation among children / 中国学校卫生

Objective@#To investigate the role of miR-142-3p in alleviation of house dust mite induced allergic airway inflammation among children, so as to provide insights into unraveling the pathogenesis of allergic airway inflammation.@*Methods@#Serum samples were collected from 15 patients with house dust mite induced allergic asthma and 15 healthy children in Jiangnan University Medical Center from September to November 2022, and serum miR-142-3p expression was quantified using a fluorescent quantitative real time PCR (qPCR) assay. The levels of interleukin 6 (IL 6) and tumor necrosis factor α (TNF α) were measured in the cell culture supernatant using enzyme linked immunosorbent assay (ELISA), and the expression of high mobility group box 1 (HMGB1) was detected at transcriptional and translational lvels using qPCR and Western blotting assays. The negative regulation of the HMGB1 gene by miR 142 3p was identified using a dual luciferase gene reporter assay, and the expression of downstream regulatory proteins was determined in human normal lung epithelial cells (BEAS 2B) cells transfected with miR 142 3p using Western blotting. In addition, female C57BL/6 mice at ages of 6-8 weeks were randomly assigned to the phosphate buffer saline (PBS) group, house dust mite sensitized airway inflammation group and house dust mite sensitized airway inflammation + miR 142 3p intervention group. Mouse airway inflammation was evaluated using hematoxylin eosin staining, and the expression of inflammatory cells and inflammatory cytokines were detected in mouse bronchoalveolar lavage fluid (BALF) using Giemsa staining and ELISA.@*Results@#Lower serum miR-142-3p expression was quantified among children with house dust mite induced allergic asthma than among healthy controls (1.33±0.21 vs. 4.74±0.62, t=5.22, P <0.05). Stimulation with dermatophagoides farinae extract (DFE) resulted in a reduction in miR-142-3p expression in BEAS-2B cells (0.82±0.25), while transfection with miR-142-3p mimics resulted in a rise in miR-142-3p expression in BEAS-2B cells (0.55±0.14)( t=3.31, 3.94, P <0.05). Pre treatment with miR-142-3p reduced the expression of IL 6(2.25±0.46)and TNF α(6.58±1.95) ( t=4.86, 3.38, P <0.05) in BEAS 2B cells stimulated with DFE, and treatment with miR-142-3p mimics resulted in a reduction in TLR4 and NF-κB expression in BEAS-2B cells via negative regulation of the HMGB1 expression. In addition, treatment with miR-142-3p was found to alleviate inflammatory cell infiltration in lung tissues of house dust mite sensitized mice, and results in a reduction in interleukin 4 (IL-4)[(107.60±10.43)pg/mL], interleukin 5 (IL 5)[(95.78±13.14)pg/mL] and HMGB1[(2.52±0.87)pg/mL] expression in BALF ( t=10.32, 7.29, 2.90, P <0.05).@*Conclusion@#miR-142-3p alleviates house dust mite induced allergic airway inflammation among children via negative regulation of the HMGB1/TLR4/NF-κB pathway.

Bacterial community diversity in Dermatophagoides farinae using high-throughput sequencing / 中国血吸虫病防治杂志

Objective To investigate the bacterial community diversity in Dermatophagoides farinae. Methods Laboratory-cultured D. farinae was collected, and the composition of microbial communities was determined by sequence analyses of the V4 region in the bacterial 16S ribosomal RNA (16S rRNA) gene on an Illumina PE250 high-throughput sequencing platform. Following quality control and filtering of the raw sequence files, valid reads were obtained and subjected to operational taxonomic units (OTU) clustering and analysis of the composition of microbial communities and alpha diversity index using the Usearch software, Silva database, and Mothur software. Results A total of 187 616 valid reads were obtained, and 469 OTUs were clustered based on a sequence similarity of more than 97%. OTU annotation showed that the bacteria in D. farinae belonged to 26 phyla, 43 classes, 100 orders, 167 families and 284 genera. The bacteria in D. farinae were mainly annotated to five phyla of Proteobacteria, Firmicutes, Bacteroidota, Actinobacteriota, and Acidobacteriota, with Proteobacteria as the dominant phylum, and mainly annotated to five dominant genera of Ralstonia, norank-f-Mitochondria, Staphylococcus and Sphingomonas, with Wolbachia identified in the non-dominant genus. Conclusions A high diversity is identified in the composition of the bacterial community in D. farinae, and there are differences in bacterial community diversity and abundance among D. farinae.

Scanning electron microscopic observation of the external morphology of Dermatophagoides farinae at different developmental stages / 中国血吸虫病防治杂志

Objective To investigate the morphological characteristics of Dermatophagoides farinae at different developmental stages. Methods The cultured D. farinae was isolated, and the external morphological features of mites at various developmental stages were observed using scanning electron microscopy (SEM), including egg, larva, nymph and adult stages. Results The D. farinae egg appeared a long oval shape, and the larval mites had three pairs of legs. The nymph had four pairs of legs and underdeveloped genital pores containing genital setae and anal setae, and adult mites appeared long and oval in shape, with decorative patterns on epidermis, and had four pairs of legs. In male adult mites, remarkable thickening of the leg I and thicker and longer leg III than the leg IV were seen, and ventral genital regions were found between the basal segments of legs III and IV; the anus was surrounded by a circular peri-anal ring, with a pair of anal suckers and anal setae within the ring. In the female adult mites, slender legs III and IV with an equal length were seen, and a “λ-shape” genital hole was observed on the ventral surface, with a crescent-like genital plate in the anterior part, and the anus appeared a longitudinal slit. Conclusions An SEM observation of the external morphology of D. farinae provides understandings of the morphological characteristics of D. farinae, which is of great significance for the classification and identification.

Cloning and sequence analysis of leptin receptor overlapping transcript-like 1 gene from Dermatophagoides farinae / 中国血吸虫病防治杂志

Objective To obtain the leptin receptor overlapping transcript-like 1 encoding gene (LepROTL1 gene) from Dermatophagoides farina, investigate the molecular characteristics of the gene and construct a prokaryotic expression vector to express this gene. Methods The LepROTL1 gene-encoding sequence fragments were captured based on the transcriptome sequencing results, and the full-length gene fragments were amplified from total RNA of D. farinae using a RT-PCR assay, and used to construct the expression plasmid pET28a(+)-LepROTL1, followed by sequencing. The plasmid was transformed into E. coli BL21 (DE3) T1R for the induction of IPTG expression. The expression product was characterized by SDS-PAGE and Western blotting. Bioinformatics analyses were performed to analyze the sequence and the molecular characteristics of its encoded protein. Results The amplification products of the RT-PCR assay showed a clear band on agarose gel electrophoresis, and sequencing analysis of the pET28a(+)-LepROTL1 plasmid showed 417 bp in length of the coding gene from the start codon ATG to the termination codon TAA. Following the plasmid transformation into E. coli and induction with IPTG, a specific band was seen on SDS-PAGE, indicating successful expression. Bioinformatics analysis showed that the LepROTL1 gene-encoded protein was composed of 134 amino acids, and had a relative molecular weight of 14 378.13 Da, a hydrophilicity index of 1.149, and certain hydrophobicity. The secondary structure was composed of alpha-helix (19 aa, 14.18%), extended strand (48 aa, 35.82%) and random coil (67 aa, 50.00%). The deduced amino acid sequence was used to obtain homologous genes by BLAST, and the phylogenetic tree showed that D. farinae was clustered with D. pteronyssinus. Conclusion The full-length sequences and expression plasmid of the LepROTL1 gene are obtained, and the molecular features of the gene are demonstrated using bioinformatics analyses, which provide insights into further studies on the gene.

Performance of magnetic nanoparticle chemiluminescence immunoassay in detection of allergen-specific immunoglobulin E / 第二军医大学学报

Objective To evaluate the performance of magnetic nanoparticle chemiluminescence immunoassay (NM-CLIA) in detection of allergen-specific immunoglobulin E (sIgE) antibodies to Dermatophagoides pteronyssinus (International Allergen Code D1) and Dermatophagoides farinae (International Allergen Code D2). Methods A total of 489 serum samples from the patients with suspected allergic disease (244 cases caused by D1, and 245 caused by D2), who were treated at Suzhou Kowloon Hospital, Shanghai Jiaotong University School of Medicine, were detected by NM-CLIA and immunofluorescence assay, respectively. χ2 test and Kappa test were used to evaluate the correlation between the two methods in detection of D1 and D2 sIgE antibodies. The limit of detection (LoD), linear range and precision of NM-CLIA in detection of D1 and D2 sIgE antibodies were verified by the standard method of American Clinical Laboratory Standardization Association. Results The LoDs of NM-CLIA in detecting D1 and D2 sIgE antibodies were both less than 0.01 U/mL, the linearity ranged from 0.1 to 100 U/mL, the within-run precision was less than 5%, and the between-run precision was less than 8%. Methodological comparison results showed that NM-CLIA and immunofluorescence assay had good consistency in detecting D1 and D2 sIgE antibodies. For D1, the positive coincidence rate and negative coincidence rate were 95% and 92%, respectively (χ2＝174.45, P0.001, Kappa＝0.843), and the ±1 class agreement was 95.6%; for D2, the positive coincidence rate and negative coincidence rate were 91% and 97%, respectively (χ2＝154.263，P0.001，Kappa＝0.787), and the ±1 class agreement was 94.2%. Conclusion NM-CLIA has good correlation with immunofluorescence assay in detecting D1 and D2 sIgE antibodies, and has good LoD, linear range and precision, suggesting that it can be recommended for clinical testing of D1 and D2 sIgE antibodies.

Evaluation of efficacy of 1 year after completing the 2-year specific sublingual immunotherapy treatment course in allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

Objective@#To evaluate the efficacy of treatment at 1 year after completing the 2-year specific sublingual immunotherapy in patients with allergic rhinitis (AR).@*Methods@#A retrospective analysis of 124 patients with AR who were allergic to dust mite and treated in the First Affiliated Hospital of Wenzhou Medical University since 2012 was performed. All patients achieved sublingual immunotherapy (SLIT) for 2 years and had a regular follow-up over 1 year. Efficacy evaluation index such as VAS score, total nasal symptom score and total medication score were compared between the time of SLIT for 2 years, and 1 year after termination of SLIT. Paired t test was used for the comparison of scores.@*Results@#Compared with those before treatment, the VAS score, total nasal symptom score and total medication score at the time of SLIT for 2 years were significantly decreased in all patients with AR [(9.40±5.96) vs (24.78±6.36), (4.53±2.06) vs (9.51±2.02), (0.42±0.87) vs (3.02±0.41), t value was 17.627, 24.600, 5.331, respectively, all P<0.01]. Compared with 2 years SLIT treatment, VAS score and total nasal symptom score were significantly increased than 1 year after termination of immunotherapy [(12.52±6.92) vs (9.40±5.96), (5.30±2.36) vs (4.53±2.06), t=5.199, 3.744, respectively, all P<0.01], but the total medication score showed no significant difference [(0.34±0.84) vs (0.42±0.87), t=-1.043, P>0.05].@*Conclusions@#The symptoms after 2 years SLIT in patients with AR have been well controlled. One year after the termination of SLIT, there is a certain rebound in symptoms, 2 years SLIT may not be the ideal treatment cycle for AR.

Therapeutic effects of orally administered CJLP55 for atopic dermatitis via the regulation of immune response

Atopic dermatitis (AD) is an inflammatory skin condition accompanied by symptoms such as edema and hemorrhage. Kimchi is a traditional fermented Korean dish consisting of various probiotics. In this study, the therapeutic effect of Lactobacillus plantarum CJLP55 isolated from Kimchi was studied in AD-induced mice. Orally administered Lactobacillus strain, CJLP55, suppressed AD symptoms and high serum IgE levels. CJLP55 administration reduced the thickness of the epidermis, infiltration of mast cells and eosinophils into the skin lesion, enlargement of axillary lymph nodes, and increase in cell population in axillary lymph nodes. CJLP55 treatment decreased the production of type 2 cytokines, such as interleukin (IL)-4, IL-5, IL-10, IL-12, interferon (IFN)-γ, and IL-6,which were stimulated by house dust mite extracts, in the axillary lymph node cells. Orally administered CJLP55 exhibited a therapeutic effect on house dust mite-induced AD in NC/Nga mice after onset of the disease by altering immune cell activation. The Lactobacillus strain, CJLP55, isolated from Kimchi, suppressed AD. Our results suggest its possible use as a potential candidate for management of AD.

House Dust Mite Sensitization Is Inversely Associated with Plasma 25-Hydroxyvitamin D3 Levels in Patients with Severe Atopic Dermatitis

BACKGROUND: The relationship between atopic dermatitis (AD) and low vitamin D levels has been studied. Emerging evidence has implicated vitamin D as a critical regulator of immunity, playing a role in both the innate and cell-mediated immune systems. However, the effect of vitamin D on house dust mite (HDM) sensitization in patients with AD has not been established. OBJECTIVE: We investigated the association between vitamin D levels and HDM sensitization according to AD severity. METHODS: In total, 80 patients (43 men and 37 women) with AD were included. We classified AD severity using Rajka and Langeland scores. Laboratory tests included serum 25-hydroxyvitamin D3, total immunoglobulin E (IgE), and specific IgE antibody titer against Dermatophagoides farinae and D. pteronyssinus. RESULTS: There were no differences in vitamin D levels between the mild or moderate AD and severe AD groups. In the severe AD group, high HDM sensitization group had lower serum vitamin D levels compared to low HDM sensitization group with statistical significance. In addition, a significant negative correlation was found between vitamin D levels and HDM sensitization in the severe AD group. CONCLUSION: Our results demonstrate that low vitamin D levels may link to high HDM sensitization in patients with the severe AD. Further elucidation of the role of vitamin D in HDM sensitization may hold profound implications for the prevention and treatment of AD.

Usefulness of House Dust Mite Nasal Provocation Test in Asthma

PURPOSE: We previously reported that the skin prick test was sensitive and the serum specific immunoglobulin E test was specific for predicting positive airway responses to house dust mites (HDMs) in patients with asthma. Because the nose and bronchus are one airway, the nasal provocation test would be more specific for predicting the bronchial responses to HDM than the skin test. METHODS: The allergy skin prick test and nasal and bronchial provocation tests using HDM (Dermatophagoides farinae) were performed in 41 young men (age, 19–28 years) who wanted military certification for asthma. The nasal responses to HDM was scored according to the severity of rhinorrhea, sneezing, and nose itching. RESULTS: The prevalence of a positive skin prick test to HDM did not significantly differ between patients with (n=24) and without (n=17) an early airway reaction (EAR; 79.2% vs 70.6%, P=0.534). However, the prevalence of a positive nasal test was significantly higher in the airway responders than in the others (37.5% vs 0%, P=0.005). The concordance of a positive response to the nasal test (κ=0.332, P=0.004) but not to the skin prick test (κ=0.091, P=0.529) was significant with an EAR. The diagnostic sensitivity of the nasal test (37.5%) was lower than that of the skin prick test (79.2%), but the specificity was higher (100% vs 29.4%). CONCLUSIONS: The skin prick test is more sensitive, whereas the nasal test is more specific and accurate, for predicting an EAR to HDM in patients with asthma.

Efficacy evaluation of specific sublingual immunotherapy with standardized dust mites drops on different age patients with allergic rhinitis / 中国耳鼻咽喉头颈外科

OBJECTIVE To evaluate the difference of efficacy of sublingual immunotherapy on different age patients with allergic rhinitis. METHODS 229 patients with dermatophagoides farina drops allergic rhinitis who have finished SLIT at least 2 years were enrolled and analyzed by retrospective analysis. All patients were divided into different age groups: pre-adolescence(n=107), adolescence(n=53), adulthood(n=69). Indexes for therapeutic effects of SLIT visual analogue scale(VAS) score, rhinitis symptom scores, medication scoreintra- and inter groups were assessed during different stage of treatment(3 months, 1 year and 2 years). Data were analyzed with SPSS 19.0 software. RESULTS Compared with prior treatment, after the SLIT for 3 months, 1 year, 2 years, VAS scores, rhinitis symptom scores, medication scoresof all the patients with allergic rhinitis are significantly decreased; Differences of rhinitis symptom scores(H=0.844, 4.153, 2.669, P>0.05), VAS score(H=1.356, 3.720, 0.313, P >0.05), medication scores(H=1.044, 5.841, 3.399, P>0.05) between groups had no statistical significance at different stage after treatment(3 months, 1 year and 2 years); The differences between 2 years and 1 year of SLIT treatment showed statistical significance in the VAS scores(Z=1.635, P =1.635). CONCLUSION SLIT with dust mites drops had good curative effect and security in patients with allergic rhinitis, which hadno obvious difference between different age groups.

Short-term clinical efficacy of sublingual immunotherapy in children with allergic rhinitis / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the short-term clinical efficacy of sublingual immunotherapy in children with allergic rhinitis. METHODS Forty-four patients, from 3-14 years of age, with mite-sensitive allergic rhinitis were treated sublingually with standardized dermatophagoidesfarinae extract. After treatment of 3 or 6 months using the dermatophagoidesfarinae extract, the total nasal symptoms score(including nasal congestion, nasal itching, rhinorrhea and sneezing scores), total rhinitis medicine scores, visual analog scale scores and the percentage of blood eosinophilic granulocyte(Eos%) before and after treatment were evaluated. SPSS 20.0 software was used to analyze the data. RESULTS After 3 or 6 months' treatment,thenasal symptoms score(including nasal congestion, nasal itching, rhinorrhea and sneezing scores), total rhinitis medicine scores and visual analog scale scoreshad significantly reduced,comparing with that before treatment(all P0.05), while nasal itching scores had significantly reduced(Z=-4.185, P<0.05). After 6 months' treatment, the percentage of Eos% had significantly reduced compared with that before treatment(Z=-4.544, P<0.05). CONCLUSION SLIT could be an effective treatment in patients with allergic rhinitis sensitive to house dust mite. It would have effect after3 months treatment and Eos%reduced significantly after 6 months treatment.

Therapeutic Effects of Bailing Capsules Combined with Dermatophagoides Farinae Drop in the Treatment of Allergic Rhinitis / 中国药师

Objective:To investigate the clinical effects of dermatophagoides farinae drop combined with Bailing capsules in the treatment of allergic rhinitis. Methods:Totally 96 patients with allergic rhinitis in our hospital were selected and randomly divided into two groups with 48 cases in each by a randomization table. The patients in the treatment group received dermatophagoides farinae drop and Bailing capsules,and those in the control group only received dermatophagoides farinae drop. The scores of symptoms and physical signs and adverse reactions were compared between the two groups. Results:After the treatment,the scores of symptoms and physical signs in the two groups were significantly decreased(P 0. 05). Conclusion:Bailing capsules combined with dermatophagoides farinae drop can significantly relieve the clinical symptoms and signs,and improve the quality of life of the patients with allergic rhinitis,which is worthy of promoted application in clinics.

Synergistic Effect of Dermatophagoides farinae and Lipopolysaccharides in Human Middle ear Epithelial Cells

PURPOSE: Although the concept of "one airway, one disease," which includes the middle ear space as part of the united airway is well recognized, the role of allergens in otitis media with effusion (OME) is not clearly understood. We aimed to investigate the effect of the interaction between Dermatophagoides farinae (Der f) and lipopolysaccharide (LPS) on the induction of epithelial inflammatory response in vitro. METHODS: Primary human middle ear epithelial cells were exposed to Der f, LPS, or both in different sequences, and the magnitude of the immunologic responses was compared. The mRNA expressiona of mucin (MUC) 4, 5AC, 5B, 8, GM-CSF, TNF-α, TLR4, and MD-2 were evaluated using real-time PCR. MUC levels before and after siRNA-mediated knockout of TLR4 and MD-2 were assessed. Lastly, the involved cell signaling pathway was evaluated. RESULTS: The expressiona of cytokines, and the MUC 4, 5AC, 5B, and 8 genes were augmented by pretreatment with Der f followed by LPS; however, reverse treatment or combined treatment did not induce the same magnitude of response. Increased MUC expression was decreased by TLR4 knockdown, but not by MD-2 knockdown. The signal intensity of MUC 8 was higher in MD-2 over-expressed cells than in those exposed to LPS only. The translocation of nuclear factor-κB was observed in cells pretreated with Der f followed by LPS. CONCLUSIONS: When Der f treatment preceded LPS exposure, Der f and LPS acted synergistically in the induction of pro-inflammatory cytokines and the MUC gene, suggesting an important role in the development of OME in patients with concealed allergy airway sensitization.

Comparison of IgE-binding components between 2 house dust mites in adult allergic patients

PURPOSE: This study investigated the differences in the profile of IgE-binding components between Dermatophagoides pteronyssinus (Dp) and Dermatophagoides farina (Df) in respiratory allergic patients sensitized to Dp/Df. METHODS: Eighteen patients with respiratory allergic diseases having higher levels of serum specific IgE to Df compared to those to Dp (>twice) were enrolled. IgE-immunoblot analysis using Dp and Df extracts were used to compare the IgE binding components. Study subjects were classified into 2 groups according to the results of IgE-immunoblot analysis: 6 subjects having IgE-binding components to group 1 and 2 allergens (group B) and 12 subjects not having them (group A). RESULTS: Group A subjects were older (47.92±8.51 vs. 35.50±11.10, P=0.039) and males were dominant (75% vs. 0% P=0.009). IgE-immunoblot analysis demonstrated that all the group B subjects had IgE bindings to 2 major components, 14 and 25 kDa, while group A subjects had IgE bindings to high-molecular weight components ranging from 60-98 kDa. The enzyme-linked immunosorbent assay inhibition test showed a significant inhibition with additions of Df, not with Dp in group B subjects. Serum specific IgE levels to Dp and Df were significantly higher in group B than in group A, while its ratio (Df to Dp) was significantly higher in group A. No differences were noted in clinical parameters, total IgE, or eosinophil cationic protein levels. CONCLUSION: Heterogeneity of IgE binding patterns to Dp and Df extracts was noted according to the ratio of serum specific IgE (Df/Dp).

Comparison of short-term effects between subcutaneous and sublingual immunotherapies in children with house dust mite-sensitized allergic rhinitis and asthma

PURPOSE: There have been several studies on comparisons of efficacy between subcutaneous immunotherapy (SCIT) and sublingual immunotherapy (SLIT) in children with house dust mite (HDM)-sensitized allergic rhinitis (AR), without consistent results. This study was conducted to compare short-term effects between SCIT and SLIT in Korean children with HDM-sensitized AR. METHODS: Fifty-three children (mean age, 11.15+/-2.82 years) with HDM-sensitized AR and with/without asthma (SCIT group, n=33; SLIT group, n=20) were enrolled. Clinical symptom scores and skin prick test results were assessed before, and after 3, 6, and 12 months of immunotherapy. Blood tests, including eosinophils, total serum IgE, and HDM-specific IgE, and adenosine 5'-monophosphate, and methacholine bronchial challenge tests were performed before and after 12 months of immunotherapy. RESULTS: In the SCIT group, the symptom scores improved after 3 months compared to those before immunotherapy, whereas they improved after 6 months in the SLIT group. Significant decreases in skin reactivity to HDM were observed after 3 months only in the SCIT group. Decreases in total eosinophil counts and improvements in methacholine bronchial provocation tests were observed after 12 months of immunotherapy only in the SCIT group. No difference in severe adverse reactions was noted in either group. CONCLUSION: The results of this study suggest that SCIT may have more rapid effects on clinical symptoms and skin reactivity in children with AR, compared to SLIT.

Actividad acaricida del aceite esencial de la corteza de Croton malambo H. Karst, metil-eugenol y metil-isoeugenol contra Dermatophagoides farinae Hughes, 1961 / Acaricide activity of essential oil of Croton malambo H. Karst, methyl-eugenol and methyl-isoeugenol against Dermatophagoides farinae Hughes, 1961

Dermotophagoides farinae lives in the indoor environment of houses, where it is source of allergens; therefore its control is a priority in preventing respiratory allergies. The aim of this study was to evaluate the acaricidal activity of essential oil of Croton malambo H. Karst bark and their components methyl-eugenol and methyl-isoeugenol against Dermatophagoides farinae. The essential oil was obtained through hydro-distillation assisted by microwave radiation and analyzed by GC-MS. Acaricidal activity was assessed by dose- response bioassay, at different times, using benzyl benzoate as a positive control. The relative amount of methyl-eugenol and methyl- isoeugenol in the essential oil was 68.4 percent and 4.9 percent, respectively. The acaricidal activity was: essential oil > methyl-eugenol > benzyl benzoate > methyl-isoeugenol. The acaricidal properties of essential oil of Croton malambo bark and methyl-eugenol against Dermatophagoides farinae were established...

Dermatophagoides farinae habita en el interior de las casas, donde es fuente importante de alérgenos, por tanto su control es una prioridad en la prevención de alergias respiratoria. El objetivo de este estudio fue evaluar la actividad acaricida del aceite esencial de la corteza de Croton malambo H. Karst y sus componentes metil-eugenol y metil-isoeugenol contra Dermatophagoides farinae. El aceite esencial se obtuvo por hidrodestilación asistida por radiación de microondas y se analizó por CG-EM. La actividad acaricida se evaluó mediante bioensayo de dosis-respuesta a diferentes tiempos, usando benzoato de bencilo como control. La cantidad relativa de metil-eugenol y metil-isoeugenol en el aceite fue 68.4 por ciento y 4.9 por ciento, respectivamente. La actividad acaricida fue: aceite esencial > metil-eugenol > benzoato de bencilo > metil-isoeugenol. Se establecieron las propiedades acaricidas del aceite esencial de la corteza de Croton malambo y metil-eugenol contra Dermatophagoides farinae...

Impacto de la inmunoterapia subcutánea con Dermatophagoides farinae y Dermatophagoides pteronyssinus sobre la calidad de vida de pacientes con rinitis y asma alérgica / The impact of subcutaneous immunotherapy with Dermatophagoides farinae and Dermatophagoides pteronyssinus on the quality of life of patients with allergic rhinitis and asthma

Introducción. Como sucede en otras partes del mundo, la prevalencia de asma y rinitis alérgica en Colombia está en aumento. Se ha establecido que la inmunoterapia subcutánea con alérgenos es eficaz a largo plazo en pacientes con rinitis alérgica y asma sensibilizados a Dermophagoides. Objetivo. Proveer evidencia sobre los cambios relacionados con la calidad de vida inducidos por la inmunoterapia subcutánea en sujetos con alergia respiratoria. Materiales y métodos. Se seleccionaron 76 sujetos con diagnóstico de alergia respiratoria con sensibilización a Dermatophagoides farinae y Dermatophagoides pteronyssinus . Para la evaluación de la calidad de vida se emplearon los instrumentos Kidscreen-27 y SF-36 ( Short form 36 ). Estos instrumentos se aplicaron en dos ocasiones: durante la primera visita, en la cual se iniciaba la inmunoterapia subcutánea, y un año después de haberse iniciado el tratamiento. Resultados. Al año de estar recibiendo la inmunoterapia, los 22 sujetos que completaron el estudio presentaron cambios positivos en términos de calidad de vida. En los niños, el principal cambio se presentó en el dominio del ´entorno escolar´ mientras que en los adultos fue en el de la ´función física´ . Discusión. Se evaluaron por primera vez en Colombia los beneficios inducidos por la inmunoterapia subcutánea para ácaros de polvo en la calidad de vida de sujetos con rinitis alérgica y asma mediante los cuestionarios Kidscreen-27 y SF-36. Los resultados proveen evidencia de que la inmunoterapia subcutánea influye positivamente en la calidad de vida en sujetos con rinitis asmática y asma sensibilizados a los ácaros de polvo.

Introduction: The prevalence of asthma and allergic rhinitis in Colombia is increasing at the same rate as it is in other parts of the world. It has been determined that allergen-specific subcutaneous immunotherapy is effective in subjects with allergic rhinitis and asthma that are sensitized to house dust mites: Dermatophagoides farinae and Dermatophagoides pteronyssinus . Objective: To provide evidence on changes in the quality of life of subjects induced by allergen-specific subcutaneous immunotherapy with Dermatophagoides farinae and Dermatophagoides pteronyssinus . Materials and methods: We selected 76 subjects with a diagnosis of respiratory allergy with sensitization to Dermatophagoides farinae and Dermatophagoides pteronyssinus . The instruments used for evaluating the quality of life were Kidscreen-27 and SF-36. These instruments were applied twice for each subject: once during the first visit, and during the twelfth visit corresponding to the one-year follow-up. Results: Twenty-two subjects completed this study. After one year of treatment with allergen-specific subcutaneous immunotherapy, we found positive changes in terms of the quality of life. In children, the main change was in the School Environment domain while in adults it was in the Physical Function domain. Conclusion: We evaluated, for the first time in Colombia, benefits induced by allergen-specific subcutaneous immunotherapy for dust mites in terms of quality of life in subjects with allergic rhinitis and asthma. These results demonstrated that allergen-specific subcutaneous immunotherapy produces a positive influence on subjects sensitized to dust mites that received allergen-specific subcutaneous immunotherapy to Dermatophagoides farinae and Dermatophagoides pteronyssinus after one year.

Cloning, expression,purification and immunological identification of Der f 11, paramyosin,a recombinant allergen of dust mite / 中国免疫学杂志

Objective: To clone , express and purify of Dermatophagoides farinae ( Der f 11 ) , and then test its immunogenicity.Methods:The gene coding for Der f 11 was synthesized ,and was then linked with the pET-32a vector.The expression plasmid pET32a(+)-Der f 11 was induced by IPTG.After purification of recombinant allergens Der 11 proteins through the Ni +affinity chromatography ,immunological allergic patients serum as the Primary antibody.Results: We obtain high purity recombinant Der f 11 protein.The results of SDS-PAGE show that the expression product is about 118 KD.Recombinant allergen Der f 11 test 15 dust mites allergic patients serum specific IgE , positive rate was 20%.Conclusion: Recombinant allergen Der f 11 obtained has the similar immunologic activity to natural Der f 11 protein.It can lay the foundation for the specific diagnosis ,treatment and further experimental studies of the dust mite allergy disease.

Observation of mast cells and detection of related cytokines in murine bronchial asthma model pre and post desensitization therapy / 中国免疫学杂志

Objective:To prepare Dermatophagoides farinae (Der f) crude protein to establish BALB/c bronchial asthma model , and to observe the morphology and degranulation of mast cells and detect related cytokines .Methods: Dermatophagoides farinae ( Der f) crude protein were prepared by trituration .30 BALB/c mice were randomly divided into 3 groups:PBS control group (A), asthma model group (B) and Der f crude protein treatment group (C).Group A were treated with PBS(100 μl) all the time, group B and group C were treated with 50 μg Der f crude protein mixed with 50μl alum adjuvant on day 0,day 7 and day 14.On day 28 group A and B were subcutaneous injected with PBS (100 μl) and group C were subcutaneous injected with Der f crude protein (350μg) in PBS(100 μl) at 1-day intervals.One week after the last treatment ,group A,B and C were intranasally challenged with 50 μg Der f crude protein daily for seven days .Twenty-four hours after the last challenge , airway hyper-responsiveness ( AHR) was assessed by using whole-body plethysmography .Two days post challenged , mice were sacrificed and bronchoalveolar lavage fluid ( BALF) was collected.Number of the total cells and eosinophil was determined .Level of IL-4,IL-10 and IFN-γcytokines in the BALF and the su-pernatant of splenocyte culture was assayed by ELISA .Level of Der f specific IgE and histamine in the sera was determined by ELISA . Airway inflammation was analyzed by HE staining .Observation of the morphology and degranulation of mast cells was analyzed by tolui -dine blue staining.Results:Compared with group B,AHR and the lung inflammation in group C were greatly reduced (P<0.01). Numbers of total cells and eosinophils in BALF of group C were significantly lower than that of group B ( P<0.01 ) .Compared with group B, the observation of degranulation of mast cells was insignificant in group C .Compared with group B(IgE:1.905), the level of specific IgE was significantly lower in groups C (IgE:1.278)(P<0.01).The level of IL-4 in BALF of group C was significantly lower than that of group B(P<0.01).Compared with group A and B, the level of IL-10 in BALF was significantly higher in group C (P<0.01) and the level of IFN-γin BALF of group C was significantly higher than that of group A and B (P<0.01).Compared with group B, the level of IL-4 in cultured splenocytes was significantly lower in group C (P<0.01), and the level of IL-10 and IFN-γin cultured splenocytes of group C was significantly higher than that of group B (P<0.01).Compared with group B, the level of histamine in BALF was slightly lower in groups C (P<0.05), and the level of histamine in sera was significantly lower in groups C (P<0.05). Conclusion:The degranulation of mast cells of murine bronchial asthma model was suppressed after the desensitization therapy .