Association of Single Nucleotide Polymorphisms in the MD-2 Gene Promoter Region With Der p 2 Allergy

En-Chih Liao,Ching-Yun Chang,Chia-Che Wu,Gou-Jen Wang,Jaw-Ji Tsai 대한천식알레르기학회 2015 Allergy, Asthma & Immunology Research Vol.7 No.3

Purpose: Sensitization to house dust mite (Dermatophagoides pteronyssinus) is a considerable risk factor for the progression of allergic disease. The group 2 allergen from Dermatophagoides pteronyssinus, Der p 2, is considered a major one in patients with specific immunoglobulin E (IgE) to Der p 2. Der p 2 has structural homology with myeloid differentiation 2 (MD-2), which is involved in the lipopolysaccharide-binding component of the Toll-like receptor 4 signaling pathway and the development of inflammation. The aim of this study was to examine the genetic association of single nucleotide polymorphisms (SNPs) in the promoter region of MD-2 with Der p 2-sensitive allergy. Methods: We investigated associations between cohort’s characteristics, including 280 allergic and 80 healthy subjects by examining total IgE, eosinophils, D. pteronyssinus- specific IgE, Der p 2-specific IgE, the number of IgE-producing B cells induced by Der p 2, and the odds ratio of allergic symptoms. Results: Based on the 1,000 genome project data, the minor allele frequencies of the rs1809441 and rs1809442 are 0.467 and 0.474, respectively. However, the correlation of linkage disequilibrium (LD) between these 2 SNPs is D’=1, the genotype frequencies of the 2 MD-2 (LY96) SNPs (rs1809441 and rs1809442) that are located nearby were significantly different between allergic and health subjects: the TT genotype of rs1809441 and the GG genotype of rs1809442 were more frequent in allergic subjects than in healthy subjects (16.1% vs 2.5% in both genotypes). The allergic patients with these genotypes exhibited significantly higher levels of D. pteronyssinus-specific IgE and Der p 2-specific Ig E, and a larger number of Der p 2-activated B cells. In addition, these 2 SNPs in the MD-2 promoter region were significantly associated with the prevalence of nasal, skin, and asthmatic allergic symptoms. Conclusions: Our results indicated that 2 SNPs in the MD-2 promoter region were significantly associated with Der p 2-specific Ig E, and thereby suggest that these SNPs may play a major role in susceptibility to Der p 2-triggered immune responses in a Taiwanese population.

Enhanced Allergic Inflammation of Der p 2 Affected by Polymorphisms of MD-2 Promoter

En-Chih Liao,Chia-Wei Hsieh,Ching-Yun Chang,Sheng-Jie Yu,Meei-Ling Sheu,Sheng-Mao Wu,Jaw-Ji Tsai 대한천식알레르기학회 2015 Allergy, Asthma & Immunology Research Vol.7 No.5

Purpose: Myeloid differentiation-2 (MD-2) has been associated with endotoxin and inflammatory disorders because it can recognize lipopolysaccharide (LPS) binding and attenuate Toll-like receptor 4 (TLR4)-mediated signaling. However, its role in allergic inflammation has yet to be clarified. We examined whether single nucleotide polymorphisms (SNPs) in MD-2 promoter can affect MD-2 expression and aimed to clarify the relationship between Der p 2 allergy and SNPs of MD-2 promoter. Methods: The function of SNPs of MD-2 promoter and the effects of cytokines and immunoglobulin on the secretion and mRNA expression were investigated in 73 allergic subjects with different MD-2 gene promoter variants. Peripheral blood mononuclear cells were cultured with or without LPS in the presence of Dermatophagoides pteronyssinus group 2 allergen (Der p 2), followed by mRNA extraction and cytokine expression analysis. The culture supernatants were collected for cytokine measurement. Results: Patients with the MD-2 promoter SNPs (rs1809441/rs1809442) had increased mRNA expressions of MD-2, ε heavy chain of IgE (Cε), and interleukin (IL)-8; however, only MD-2 and IL-8 were further up-regulated after Der p 2 stimulation. Patients with SNPs of MD-2 promoter tended to have high levels of IL- 1β, IL-6, IL-8, IL-10, and tumor necrosis factor (TNF)-α after Der p 2 and LPS stimulation. Increased secretions of IL-6, IL-8, and IL-10 were found to be up-regulated by Der p 2 stimulation, and an increased secretion of IFN-γ and decreased secretion of IL-4 were noted after LPS stimulation. Conclusions: The high levels of proinflammatory cytokines secreted by Der p 2 were predetermined by MD-2 promoter SNPs (rs1809441/rs1809442). Through cytokine secretion by Der p 2 and LPS, these SNPs may serve as an indicator of the pathological phenotype of Der p 2-induced allergic inflammation.

Dynamic Route Choice Behaviour and Simulation-Based Dynamic Traffic Assignment Model for Mixed Traffic Flows

Ta-Yin Hu,Chee-Chung Tong,Tsai-Yun Liao,Li-Wen Chen 대한토목학회 2018 KSCE JOURNAL OF CIVIL ENGINEERING Vol.22 No.2

Intelligent Transportation Systems (ITS) focus on increasing the efficiency of existing surface transportation systems through the use of advanced computers, electronics, and communication technologies. In order to perform advanced traffic management and provide travel information, dynamic traffic assignment models need to be developed to provide time-dependent estimates of traffic flows on networks in order to efficiently utilize possible advanced traffic information as well as traffic control measures. Traffic assignment distributes Origin-Destination (OD) trips in a network and determines the flow patterns in a traffic network. This research aims at developing simulation-based algorithm for dynamic traffic assignment problems under mixed traffic flow considerations. Four different physical vehicle types are explicitly considered and modeled, including car, bus, motorcycle, and truck. Four different behavioral rules, pre-specified-path driver, user-equilibrium driver, system-optimization driver, and real-time information driver, are considered in the solution procedure. The DTA algorithm consists of an inner loop that incorporates a direction finding mechanism for the search process for System Optimization (SO) and User Equilibrium (UE) classes based on the simulation results of the current iteration, including experienced vehicular trip times and marginal trip times. In order to understand tripmaker acceptance toward route guidance, a survey is conducted to explore possible behavioral classifications and associated percentages. Numerical experiments are conducted in a test network and a real city network to illustrate the capabilities of the simulation-based DTA procedures, and to observe how system performs under multiple user class’s conditions, including multiple user behavior rules and multiple physical vehicle classes.

Der p2 Internalization by Epithelium Synergistically Augments Toll-like Receptor-Mediated Proinflammatory Signaling

Sui-Chu Yin,En-Chih Liao,Chih-Liang Chiu,Ching-Yun Chang,Jaw-Ji Tsai 대한천식알레르기학회 2015 Allergy, Asthma & Immunology Research Vol.7 No.4

Purpose: House-dust-mite (HDM) major allergen Der p2 shares homology and function with Toll-like receptor (TLR) signaling protein myeloid differentiation- 2 (MD2) and may lead to airway inflammation. Should Der p2 be internalized by human airway epithelium, it has the theoretical propensity to potentiate epithelium activation. This study aimed to demonstrate the internalization of Der p2 by airway epithelium and to investigate the effects of Der p2 on MD2 expression and epithelium activation. Methods: Internalization of recombinant, enhanced green fluorescent protein-labelled Der p2 (rDer p2-EGFP) into human airway epithelium (BEAS-2B) was tracked by laser confocal microscopy and confirmed by immunoblotting. Reverse-transcription polymerase chain reaction (RT-PCR), immunoblotting, and immunohistochemical staining were used to determine the effect of Der p2 on MD2 expression in vitro and ex vivo. Expression of messenger RNA (mRNA) encoding receptors/cytokines was measured by RT-PCR. Secretion of interleukin-6/interleukin-8 (IL-6/IL-8) was measured by enzyme-linked immunosorbent assay (ELISA). Results: Internalization of Der p2 by BEAS-2B was confirmed by confocal microscopy and immunoblotting using rDer p2-EGFP and rDer p2, respectively. Expression of MD2 protein was increased in BEAS-2B and human nasal polyp airway epithelium cultured with rDer p2. Recombinant Der p2-cultured BEAS-2B caused little spontaneous IL-6/IL-8 secretion but significantly augmented by TLR ligand LPS. IL-6 secretion was up-regulated after MD2 transfection. Internalization of Der p2 was reduced by TLR2 RNA knockdown. Dexamethasone, calcitriol, anti-MD2/anti-TLR2 antibodies, and signalling inhibitors significantly reduced LPS+Der p2-induced IL-6/IL-8 secretion. Conclusions: Human airway epithelium may internalize Der p2, which potentiates the response to environmental proinflammatory stimuli through MD2 and TLRs. This study highlights a novel mechanism and alleviates IL-6/IL-8 secretion in mite-induced airway inflammation