Monotropein mitigates atopic dermatitis-like skin inflammation through JAK/STAT signaling pathway inhibition.

Atopic dermatitis (AD) is a globally increasing chronic inflammatory skin disease with limited and potentially side-effect-prone treatment options. Monotropein is the predominant iridoid glycoside in Morinda officinalis How roots, which has previously shown promise in alleviating AD symptoms. This study aimed to systematically investigate the pharmacological effects of monotropein on AD using a 2, 4-dinitrochlorobenzene (DNCB)/Dermatophagoides farinae extract (DFE)-induced AD mice and tumor necrosis factor (TNF)-α/interferon (IFN)-γ-stimulated keratinocytes. Oral administration of monotropein demonstrated a significant reduction in AD phenotypes, including scaling, erythema, and increased skin thickness in AD-induced mice. Histological analysis revealed a marked decrease in immune cell infiltration in skin lesions. Additionally, monotropein effectively downregulated inflammatory markers, encompassing pro-inflammatory cytokines, T helper (Th)1 and Th2 cytokines, and pro-inflammatory chemokines in skin tissues. Notably, monotropein also led to a considerable decrease in serum immunoglobulin (Ig)E and IgG2a levels. At a mechanistic level, monotropein exerted its anti-inflammatory effects by suppressing the phosphorylation of Janus kinase / signal transducer and activator of transcription proteins in both skin tissues of AD-induced mice and TNF-α/IFN-γ-stimulated keratinocytes. In conclusion, monotropein exhibited a pronounced alleviation of AD symptoms in the experimental models used. These findings underscore the potential application of monotropein as a therapeutic agent in the context of AD, providing a scientific basis for further exploration and development.

Clinical efficacy of Dermatophagoides farina drops on allergic rhinitis in children sensitized by dust mites combined with different allergens.

OBJECTIVE: This study aimed to investigate the clinical efficacy and safety of sublingual immunotherapy (SLIT) using standardized dermatophagoides farina drops for the treatment of allergic rhinitis (AR) in children sensitized to dust mites combined with different allergens. The findings contribute to establishing a preliminary foundation for future in-depth studies on AR treatment. METHODS: A total of 152 AR children undergoing SLIT were categorized into two groups based on serological test results: the inhalation group (dust mite combined with inhalation allergy) and the ingestion group (dust mite combined with ingestion allergy). The clinical efficacy and safety were evaluated by assessing the total nasal symptoms score (TNSS), total medication scores (TMS), visual analog scale scores (VAS scores), and the incidence of adverse reactions before treatment and after two years of treatment. RESULTS: After two years of treatment, TNSS, TMS, and VAS scores significantly improved compared to pre-treatment values in both the inhalation and ingestion groups (p < 0.05). However, there were no significant differences in efficacy between the two groups after two years of treatment (p > 0.05). During the treatment period, only 15 cases (10.9 %, 9 cases in the inhalation group and 6 cases in the ingestion group) experienced mild adverse reactions. There was no significant difference in the incidence of adverse reactions between the two groups (p > 0.05). CONCLUSION: SLIT using standardized dermatophagoides farina drops demonstrates long-term efficacy in children with AR, regardless of whether they belong to the inhalation or ingestion group, without significant differences in treatment outcomes.

Establishment of fluorescence multi-flow cytometric immunoassay for the simultaneous quantitative detection of six allergen-sIgE antibodies.

BACKGROUND: The in vitro specific IgE (sIgE) assays now commonly used in clinical laboratories are not only time-consuming and expensive but also require many serum samples. To address these limitations, a novel fluorescent microsphere-based multiplex flow cytometric immunoassay was developed. This innovative assay enables rapid and simultaneous quantitative detection of multiple allergen-sIgE antibodies. OBJECTIVE: To establish a new method for the simultaneous quantitative detection of 6 allergen-sIgE antibodies based on fluorescence multiplex flow cytometry. METHODS: Six different encoded fluorescent microspheres were selected to covalently couple 6 allergens, and their antigen-coupling activities were verified. After optimizing the multiplexing procedure and reaction conditions, including the concentration of microspheres encapsulated by allergens, reaction temperature, and reaction time, standard curves were established to quantify the 6 allergen-sIgE, and their performance was evaluated according to clinical guidelines. RESULTS: The chosen analytical mode was optimized for the detection of the 6 allergens-sIgE for 70 minutes. The established coefficients of variation for multiplex flow cytometry reproducibility and intermediate precision were less than 10%. Linear regression analysis showed a highly significant quantitative correlation between the results of the multiple analyses of Dermatophagoides pteronyssinus, Dermatophagoides farinae, Artemisia, and cat hair allergens and ImmunoCAP (Thermo Fisher Scientific): the r2 values ranged from 0.85 to 0.97 (P < .0001). In addition, there was a high correlation between the results of the multiplex analysis of dog hair allergens and the capture enzyme-linked immunosorbent assay (r2 = 0.92, P < .0001). CONCLUSION: A high-throughput system called multiplex flow cytometry has been developed for the simultaneous detection of 6 inhalant allergens. The method has the advantage of being rapid and using less serum. Furthermore, it has the potential to be expanded to include other allergens and biologic agents.

The EGR1-Artemin Axis in Keratinocytes Enhances the Innervation of Epidermal Sensory Neurons during Skin Inflammation Induced by House Dust Mite Extract from Dermatophagoidesfarinae.

Epidermal hyperinnervation is a critical feature of pruritus during skin inflammation. However, the mechanisms underlying epidermal hyperinnervation are unclear. This study investigates the role of the transcription factor EGR1 in epidermal innervation by utilizing wild-type (Egr1+/+) and Egr1-null (Egr1‒/‒) mice topically applied Dermatophagoides farinae extract from dust mite. Our findings revealed that Egr1‒/‒ mice exhibited reduced scratching behaviors and decreased density of epidermal innervation compared with Egr1+/+ mice. Furthermore, we identified artemin, a neurotrophic factor, as an EGR1 target responsible for Dermatophagoides farinae extract-induced hyperinnervation. It has been demonstrated that Dermatophagoides farinae extract stimulates toll-like receptors in keratinocytes. To elucidate the cellular mechanism, we stimulated keratinocytes with Pam3CSK4, a toll-like receptor 1/2 ligand. Pam3CSK4 triggered a toll-like receptor 1/2-mediated signaling cascade involving IRAK4, IκB kinase, MAPKs, ELK1, EGR1, and artemin, leading to increased neurite outgrowth and neuronal migration. In addition, increased expression of EGR1 and artemin was observed in the skin tissues of patients with atopic dermatitis. These findings highlight the significance of the EGR1-artemin axis in keratinocytes, promoting the process of epidermal innervation and suggesting it as a potential therapeutic target for alleviating itch and pain associated with house dust mite-induced skin inflammation.

Homology modeling and epitope prediction of Der f 33

Dermatophagoides farinae (Der f), one of the main species of house dust mites, produces more than 30 allergens. A recently identified allergen belonging to the alpha-tubulin protein family, Der f 33, has not been characterized in detail. In this study, we used bioinformatics tools to construct the secondary and tertiary structures and predict the B and T cell epitopes of Der f 33. First, protein attribution, protein patterns, and physicochemical properties were predicted. Then, a reasonable tertiary structure was constructed by homology modeling. In addition, six B cell epitopes (amino acid positions 34-45, 63-67, 103-108, 224-230, 308-316, and 365-377) and four T cell epitopes (positions 178-186, 241-249, 335-343, and 402-410) were predicted. These results established a theoretical basis for further studies and eventual epitope-based vaccine design against Der f 33.

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.

The Indoor Level of House Dust Mite Allergen Is Associated with Severity of Atopic Dermatitis in Children

We attempted to investigate the correlation between the severity of atopic dermatitis (AD) in children and the indoor level of house dust mite (HDM) allergens. Ninety-five patients (31.1 +/- 19.5 months of age) with AD were enrolled in this study, and serum specific IgE against Dermatophagoides pteronyssinus and D. farinae was measured. The severity of AD was assessed using the visual analogue scale on the same day of house dust collection. Living rooms and mattresses where the child usually slept were vacuumed for 2 minutes and concentrations of Der f 1 were measured by enzyme-linked immunosorbent assay. The skin symptoms were more severe in patients with Der f 1 concentrations in living room > 2 microg/g dust than < or = 2 microg/g dust (P = 0.018). This difference was noted in AD patients without sensitization to HDM (P = 0.004), but not in patients with sensitization. There was no difference in symptom severity according to Der f 1 concentrations in mattresses (P = 0.062). The severity of skin symptoms is associated with indoor concentrations of HDM in children with AD, and it is likely to act as nonspecific irritants as well as allergens in AD skin lesions.

Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae

Crude extracts of house dust mites are used clinically for diagnosis and immunotherapy of allergic diseases, including bronchial asthma, perennial rhinitis, and atopic dermatitis. However, crude extracts are complexes with non-allergenic antigens and lack effective concentrations of important allergens, resulting in several side effects. Dermatophagoides farinae (Hughes; Acari: Pyroglyphidae) is one of the predominant sources of dust mite allergens, which has more than 30 groups of allergen. The cDNA coding for the group 5 allergen of D. farinae from China was cloned, sequenced and expressed. According to alignment using the VECTOR NTI 9.0 software, there were eight mismatched nucleotides in five cDNA clones resulting in seven incompatible amino acid residues, suggesting that the Der f 5 allergen might have sequence polymorphism. Bioinformatics analysis revealed that the matured Der f 5 allergen has a molecular mass of 13604.03 Da, a theoretical pI of 5.43 and is probably hydrophobic and cytoplasmic. Similarities in amino acid sequences between Der f 5 and allergens of other domestic mite species, viz. Der p 5, Blo t 5, Sui m 5, and Lep d 5, were 79, 48, 53, and 37%, respectively. Phylogenetic analysis indicated that Der f 5 and Der p 5 clustered together. Blo t 5 and Ale o 5 also clustered together, although Blomia tropicalis and Aleuroglyphus ovatus belong to different mite families, viz. Echimyopodidae and Acaridae, respectively.

Usefulness of Total IgE in Predicting Positive Allergen Specific IgE Tests in Korean Subjects / 대한진단검사의학회지

BACKGROUND: Total IgE levels in allergic patients tend to be higher than those in healthy individuals. We evaluated the usefulness of total IgE levels in predicting positive results of allergen specific IgEs in multiple allergen simultaneous tests. METHODS: A total of 133 patients with allergic symptoms were evaluated. Allergen specific IgEs were detected using 3 different kits: Allergy screen (R-biopharm, Germany), AdvanSure Allergy Screen (LG Life Science, Korea) and Polycheck allergy (Biocheck Co., Germany). Total IgE was measured by turbidoimmunometric assay (LX-2200, Eiken Chemical Co., Japan). The patients were divided into high (> or =170 IU/mL) and low ( or =50%. Allergen specific IgEs to these two allergens showed good correlation with total IgE (correlation coefficients >0.5). CONCLUSIONS: Total IgE appears to be useful in predicting positive results in allergen specific IgE tests to common allergens. The specific IgEs to D. farinae and D. pteronyssinus showed good correlation with total IgE. However, for other allergens, significant differences were observed among different test kits, and the standardization of allergens in multiple allergen simultaneous tests is needed.

Determinación de sensibilización alérgica a dermatofagoides en niños de 5 años y menores por fluoroinmuno ensayo-UniCAP / Dust mite allergic sensitization in children aged 1 month to 5 years-old using specific IgE determination by unicap fluoroimmunoassay

Dust mite sensitization is considered a major risk factor for pediatric asthma; however, there are few data about early sensitization in Chilean children available. This study aimed to investigate allergic sensitization to mites in infancy and early childhood. The patient population, 224 children aged 0 to 5 years with suspected respiratory allergy, was divided into 3 groups of age: 1 to 12 months-old, 13 to 35 months-old, and 3 to 5 years-old. Sensitization status was ascertained by assessing the specific IgE to mite by UniCAP fluoroimmunoassay (Pharmacia). Sixty eight (30.6 percent) patients were sensitized to mites. The frequency and level of sensitization increased with the age of the children. Our data support that avoidance measures in the domestic environment aimed at the primary prevention of mite-driven sensitization should be introduced at the earliest possible stage.

La sensibilización a dermatofagoides es un factor de riesgo para el desarrollo de asma. Hay escasa información sobre la sensibilización temprana en niños chilenos. Objetivo: Investigar la sensibilización a dermatofagoides en niños de 5 años y menores. Método: Revisión de resultados de IgE específica por ensayo UniCAP (Pharmacia) contra dermatofagoides de niños de hasta 5 años con sospecha de alergia respiratoria, divididos en 3 grupos de edad: 1-12 meses, 13-35 meses y 3-5 años. Resultados: Se estudió un total de 224 niños, entre 1 mes y 5 años de edad (x 2,65 años +/- 1,48 DS), 57,1 por ciento de sexo masculino. Su distribución según grupo etario fue: 66 en el grupo 1; 45 en el grupo 2 y 113 en el grupo 3. El 30,6 por ciento presentó sensibilización a dermatofagoides (18,2 por ciento grupo 1; 33,3 por ciento grupo 2 y 36,3 por ciento grupo 3). El grupo 3 presentó concentraciones de IgE específica significativamente más elevadas que los otros grupos. Conclusiones: La sensibilización a dermatofagoides es detectable en menores de un año, con frecuencia y nivel que aumentan en forma paralela a la edad.

Perfil de sensibilização a alérgenos domiciliares em pacientes ambulatoriais / Indoor allergen sensitization profile in allergic patients of the allergy clinic in the University Hospital in Uberlândia, Brazil

OBJETIVO: Conhecer o perfil de sensibilização dos pacientes com diagnóstico de doenças alérgicas atendidos no Ambulatório de Alergia do Hospital de Clínicas da Universidade Federal de Uberlândia. MÉTODOS: Foi realizado um estudo retrospectivo por meio da análise de prontuários de pacientes atendidos no serviço, que foram submetidos ao teste cutâneo de puntura (TCP) para os alérgenos de Dermatophagoides pteronyssinus (Der p), Dermatophagoides farinae (Der f), Blomia tropicalis (Blo t), Canis familiaris (Can f), Felis domesticus (Fel d), Blattella germanica (Bla g) e Alternaria alternata, e respondido corretamente ao questionário ISAAC. RESULTADOS: Foram analisados 212 prontuários de pacientes que preenchiam os critérios de inclusão. A rinite alérgica isoladamente foi a maior causa de atendimento (32 por cento), seguida das associações asma e rinite (29,7 por cento) e asma, rinite e eczema (9,4 por cento). Pacientes com asma isoladamente perfizeram apenas 1,9 por cento dos atendimentos. A sensibilização dos pacientes observada foi de 73,5 por cento, sendo de 61,7 por cento para Der p, 59,9 por cento para Der f, 54,7 por cento para Blo t, 45,7 por cento para Bla g, 38,2 por cento para Can f, 33,3 por cento para Fel d e 9,9 por cento para Alternaria. Não houve diferenças significantes entre as patologias encontradas e os perfis de sensibilização. CONCLUSÃO: Os principais alérgenos sensibilizantes determinados pelo TCP foram os ácaros, com predomínio de Der p e Der f, chamando a atenção a elevada prevalência de sensibilização ao último. Ainda foi observada elevada sensibilização aos alérgenos de B. germanica, superior aos estudos anteriores realizados no País.

OBJECTIVE: To evaluate allergens among patients with allergic respiratory disease attended at the Allergy Clinic of the University Hospital - Federal University of Uberlândia. METHODS: A retrospective study was performed using medical records. Patients were included if their ISAAC questionnaires were correctly filled out and their skin prick tests were positive to at least one of the allergens from Dermatophagoides pteronyssinus (Der p), Dermatophagoides farinae (Der f), Blomia tropicalis (Blo t), Canis familiaris (Can f), Felis domesticus (Fel d), Blattella germanica (Bla g) and Alternaria alternata. RESULTS: Two-hundred and twelve medical records fulfilled the inclusion criteria. Allergic rhinitis was the main clinical diagnosis (32 percent), followed by concomitant manifestation of asthma and rhinitis (29.7 percent), and asthma, rhinitis and atopic dermatitis (9.4 percent). Asthma alone was found only in 1.9 percent of patients. The total sensitization observed was 73.5 percent, of which 61.7 percent, 59.9 percent, 54.7 percent, 45.7 percent, 38.2 percent, 33.3 percent, 9.9 percent were sensitized to Der p, Der f, Blo t, Bla g, Can f, Fel d and Alternaria, respectively. No significant difference was found between allergic disease types and source of allergen sensitization. CONCLUSION: The highest sensitization in allergic patients under study was to dust mites, especially Der p and Der f. It is noteworthy that the number of patients sensitized to cockroach extract was uncommonly high when compared to previous studies.

Relationship between serum specific IgE and allergen skin test in allergic patients of Wuhan area / 华中科技大学学报（医学）（英德文版）

In order to study the relationship between serum specific IgE (sIgE) and allergen skin test, allergen skin tests and detections of sIgE in 220 allergic patients of Wuhan area were analyzed. The coherent rate of the two methods was beyond 70% (P < 0.01). It was concluded that the in vitro and in vivo detection methods of allergens have a high coherence and can be used as the effective ways to diagnose the allergic diseases in clinical practice.

Endotoxin and House Dust Mite Allergen Levels on Synthetic and Buckwheat Pillows

Pillows are known to contain significant levels of indoor allergens and endotoxin, that are of importance to house dust mite sensitized asthmatics. Buckwheat pillows are commonly used in Korea. We studied the levels of the house dust mite allergen, Der f 1, and endotoxin on new synthetic and buckwheat pillows and their accumulation over three months. Endotoxin levels were significantly higher on new buckwheat pillows compared to synthetic pillows; geometric mean levels (95% CI) were 60,950 EU/g (30,270-122,700) and 4,887 EU/g (2,570-9,311) respectively (p<0.001). No Der f 1 was detected on the new pillows. After three months Der f 1 levels were similar on buckwheat and synthetic pillows, geometric mean levels (95% CI) were 1.16 microgram/g (0.02-8.13) and 1.08 microgram/g (0.19-1.68) respectively. These results indicate that buckwheat pillows are a source of very high endotoxin levels that may be of relevance to asthma severity of atopic asthmatics.

Der f 2 activates phospholipase D in human T lymphocytes from Dermatophagoides farinae specific allergic individuals: Involvement of protein kinase C-alpha

The major house-dust mite allergen, Der f 2, stimulates the phospholipase D (PLD) in T lymphocytes from Dermatophagoides farinae specific allergic individuals. PLD activity increased more than two-fold in T cells from allergic patients compared with those cells from normal controls with maximal responses within 30 min after exposure of Der f 2. A well-known PLD activator PKC-alpha was found to be translocated to membrane from cytosol in Der f 2-treated T cells from Dermatophagoides farinae specific allergic individuals. Down-regulation of PKC-alpha with phorbol myristate acetate pretreatment for 24 h abolished Der f 2-induced PLD activation. Ro 320432, PKC inhibitor also reduced the effects of Der f 2-induced PLD activation suggesting that PKC-alpha acts as upstream activator of PLD in Der f 2-treated T cells. Taken together, the present data suggest that Der f 2 can stimulate PLD activity through the PKC-alpha activation in T cells from Dermatophagoides farinae allergic individuals