Increased CD123 + HLA-DR- Granulocytes in Allergic Rhinitis and Influence of Allergens on Expression of Cell Membrane Markers.

BACKGROUND: It is reported that CD123 + HLA-DR- cells in PBMC are basophils, and CD203c, CD63, and FcεRI molecules are activation markers of basophils. However, little is known of CD123 + HLA-DR-cells in blood granulocytes. OBJECTIVE: To investigate the presence of CD123 + HLA-DR- cells in the blood granulocytes and peripheral PBMC of patients with allergic rhinitis (AR), as well as the impact of allergens on the cell membrane markers of basophils. METHODS: Flow cytometry was used to detect the expression of the membrane molecules. RESULTS: While CD123 + HLA-DR- PBMCs are representative of basophils, their presence did not significantly change in patients with AR. In contrast, both the percentage and number of CD123 + HLA-DR- granulocytes, which make up only up to 50% of basophils, were significantly increased in patients with seasonal (sAR) and perennial AR (pAR). CD63+, CD203c+, and FcεRIα+ cells within CD123 + HLA-DR- granulocytes also showed enhanced activity in patients with AR. Allergen extracts from house dust mite allergen extract (HDME) and Artemisia sieversiana wild extract further increased the number of CD123 + HLA-DR- cells in granulocytes of sAR and pAR patients, as well as in PBMCs of pAR patients. CONCLUSIONS: The use of CD123 + HLA-DR- granulocytes and PBMC may not be sufficient for diagnosing AR. Allergens could potentially contribute to the development of AR by influencing the number of CD123 + HLA-DR- cells, as well as the expression of CD63, CD203c, and FcεRIαin these cells.

[Detection and analysis of TLR2 expression on blood monocytes and B cells in patients with allergic rhinitis and asthma].

Objective To identify the expression of Toll-like receptor 2 (TLR2) on peripheral blood monocytes and B cells of patients with allergic rhinitis (AR), allergic rhinitis combined with allergic asthma (ARA) before and after allergen challenge. Methods The peripheral venous blood from patients with AR and ARA were recruited and stimulated with Artemisia sieversiana wild allergen extract (ASWE), house dust mite allergen extract (HDME), and Platanus pollen allergen extract (PPE). Flow cytometry was then used to detect the expression of TLR2 on peripheral monocytes and B cells. Results Compared with healthy control (HC) group, the percentage of TLR2+ monocytes and decreased mean fluorescence intensity (MFI) of TLR2 on monocytes in AR and ARA patients decreased. After being challenged with the above mentioned three allergens, the portion of TLR2+ monocytes in HC group and MFI of TLR2 on monocytes in AR patients also decreased. Meanwhile, MFI of TLR2 on B cells also showed a decrease after challenged with ASWE and HDME. Conclusion The expression of TLR2 on monocytes and B cells decreases in AR and ARA patients.

Altered Expression of Substance P and NK1R in CCR3+ and CD123+HLA-DR- Basophils Under Airway Allergic Conditions.

PURPOSE: To explore expression of SP and NK1R in basophils of allergic asthma (AA), allergic rhinitis (AR) and AR combined with AA (ARA), and influence of allergens and immunoglobulin E (IgE) mediated mechanisms on SP and NK1R expression. METHODS: Expression of SP and NK1R was detected by flow cytometry, NK1R mRNA expression was detected by real time quantitative polymerase chain reaction (qPCR), and mouse AR and AA models were employed for in vivo study. RESULTS: SP+ and NK1R+ cells increased in CCR3+ and CD123+HLA-DR- granulocytes of AA. PPE elevated proportions of SP+ cells in CCR3+ and CD123+HLA-DR- granulocytes, whereas ASWE and HDME augmented SP+ cells in CD123+HLA-DR- granulocytes of AR and ARA patients. ASWE, HDME and PPE increased proportions of NK1R+ cells in CCR3+ PBMC and CD123+HLA-DR- granulocytes of AR patients. OVA, Der p1, IL-33, IL-37, IgE and SP enhanced NK1R expression on KU812 cells. NK1R expressing basophils were increased in blood of OVA sensitized and challenged AR and AA mice. FcεRI-KO AA mice seemed to have less NK1R+ basophils than WT AA mice in their blood. CONCLUSION: CCR3+ and CD123+HLA-DR- cells are likely involved in AA and AR via SP and NK1R. IgE-related mechanism may participate in upregulation of NK1R expression.

The Transverse Bearing Characteristics of the Pile Foundation in a Calcareous Sand Area.

Reviewing literature revealed that the studies on the bearing characteristics of pile foundations mainly focuses on clay, ordinary sand, loess, saline soil, and other areas. However, few studies on the bearing characteristics of the pile foundation in calcareous sand were conducted. Besides, existing traditional studies ignored the variation of soil compression modulus with depth, and the effect of void ratio on the transverse bearing characteristics of the pile foundation in a calcareous sand area were not well understood. In response of these problems, this study conducted a theoretical investigation on the transverse bearing characteristics of the pile foundation in a calcareous sand area. The transverse bearing characteristics of the pile foundation were derived based on the Pasternak foundation model and the Winkler foundation model, incorporating the heterogeneous distribution of compressive modulus with buried depth. The calculation results of the Pasternak foundation model are closer to the observed results than the Winkler foundation model. Therefore, the following research on the transverse bearing characteristics of the pile foundation in the calcareous sand area adopts the Pasternak foundation model. Then, the effects of the pile length, pile diameter, pile elastic modulus, horizontal load, bending moment, and void ratio on the transverse bearing characteristics of the pile foundation in a calcareous sand area were thoroughly analyzed. Furthermore, the difference between the transverse bearing characteristics of the pile foundation in a calcareous sand area and a quartz sand area was discussed. Results show that the horizontal displacement of the pile top in a calcareous sand area is greater than the quartz sand area under the same conditions.

Upregulated expression of substance P and NK1R in blood monocytes and B cells of patients with allergic rhinitis and asthma.

Increased expression of substance P (SP) and neurokinin-1 receptor (NK1R) has been noticed in patients with allergic rhinitis (AR) and allergic asthma (AA). However, little is known of the expression of SP and NK1R in monocytes and B cells of AR and AA. In the present study, the expression levels of SP and NK1R were determined by flow cytometry and mouse AR and AA models. The results showed that both percentages of SP+ monocytes and SP+ B cells, and mean fluorescence intensity (MFI) of SP in monocytes were elevated in the blood of AA and AR combined with AA (ARA) patients. Similarly, the percentages of NK1R+ monocytes were elevated in the blood of AR, AA, and ARA patients. Allergens Artemisia sieversiana wild allergen extract (ASWE), house dust mite extract (HDME), and Platanus pollen allergen extract (PPE) increased the expression density of SP molecules (determined by MFI) in an individual monocyte of AR patients. HDME and PPE appeared to enhance SP and NK1R expression in the B cells of ARA and AR patients. In the mouse AR and AA models, the percentages of NK1R+ monocytes and B cells were elevated in blood following OVA (ovalbumin) sensitization and challenge. Knocking out the FcεRI molecule completely abolished the OVA-induced upregulation of expression of NK1R in monocytes and B cells of AA mice. In conclusion, upregulated expressions of SP and NK1R may contribute to the pathogenesis of airway allergy.

Increased expressions of CD123, CD63, CD203c, and Fc epsilon receptor I on blood leukocytes of allergic asthma.

Background : Altered basophil identification markers have been discovered to associate with allergic asthma (AA) in recent years. However, little is known about the expression of basophil markers in blood granulocytes. Aim: To parallel test blood basophils in peripheral blood mononuclear cell (PBMC) and granulocyte populations of patients with AA and AA combined with allergic rhinitis (ARA) Methods: The expressions of surface molecules were determined via flow cytometry. CD123 expressing cells in blood were isolated using a cell sorting technique, and mouse AA models were employed for in vivo study. Results: The numbers of CD123+HLA-DR- cells in the granulocytes of AA and ARA patients markedly increased. However, only 49.7% of CD123+HLA-DR- cells in granulocytes and 99.0% of CD123+HLA-DR- cells in PBMCs were basophils. Almost all CD123+HLA-DR- cells expressed CD63 regardless in granulocytes or PBMC. The numbers of CD63, Fc epsilon receptor I (FcεRI), and CD203c expressing cells markedly enhanced in CD123+HLA-DR- granulocytes of AA and ARA patients. Mean fluorescence intensity (MFI) of CD63 and CD203c expressions on CD123+HLA-DR- PBMC and granulocytes of AA and ARA patients dramatically elevated. House dust mite extract (HDME) and Artemisia sieversiana wild allergen extract (ASWE) enhanced the numbers of CD63+CD123+HLA-DR- granulocytes and PBMC and the MFI of CD203c expression on CD123+HLA-DR- granulocyte of AA and ARA patients. Histamine, tryptase, and PGD2 enhanced proportions of CD123+ KU812 cells. ASWE- and HDME-induced AA mice showed upregulated CD63 expression on basophils. In conclusion, upregulated expressions of CD123, CD203c, CD63, and FcεRIα in PBMC and granulocytes of patients with AA and ARA suggest that CD123+HLA-DR- cells may contribute to the development of AA and ARA.

Buckling stability analysis of underpinning piles during basement excavation beneath existing buildings.

In the present paper, the pile-soil system's total potential energy equation of underpinning piles was established based on the Winkler elastic foundation beam theory. This energy equation was used to explore the effect of basement excavation beneath existing buildings on the underpinning pile' buckling stability. Utilizing the minimum potential energy theory, the expression of the critical buckling load for underpinning piles' stability during the excavation project was obtained. Moreover, the influencing factors of the critical buckling load were investigated. It was found that the underpinning pile's critical load converged with the augment of half-wave number. Moreover, the pile skin friction and deadweight had an insignificant influence on it. In addition, the critical load of underpinning piles decreased sharply with the increasing excavation depth and gradually increased with the augment of pile diameter. The results of this study provides a basis for the design of adding piles in similar projects and reduces the hidden danger of excavation instability.

A Novel Anderson-Evans Polyoxometalate-based Metal-organic Framework Composite for the Highly Selective Isolation and Purification of Cytochrome C from Porcine Heart.

Anderson-Evans type polyoxometalate group (Na6[TeW6O24]·22 H2O, TeW6) was combined with porous metal-organic framework ZIF-8 by electrostatic interaction to obtain a novel Anderson-Evans polyoxometalate-based metal-organic framework composite, TeW6 @ZIF-8. FT-IR, Raman, XRD, TG, DSC, SEM, and TEM were used to characterize the composite. It was proved that the Anderson-Evans type polyoxometalate group TeW6 was successfully hybridized with metal-organic framework ZIF-8, and the composite possesses good stability. Based on the potential interaction between TeW6 and proteins and the coordination between imidazole groups in ZIF-8 and proteins with a porphyrin ring structure, the adsorption selectivity towards different proteins on the TeW6 @ZIF-8 composite was studied in this work. The experiment results showed that the TeW6 @ZIF-8 composite was selectively adsorbed to cytochrome C. At pH 11.0, the adsorption efficiency of 94.01% was obtained for processing 1.0 mL 100 µg mL-1 cytochrome C with 3.0 mg TeW6 @ZIF-8 composite. The adsorption behavior of cytochrome C fits well with the Langmuir adsorption model, corresponding to a theoretical adsorption capacity of 232.56 mg g-1. The retained cytochrome C could be readily recovered by 1% SDS (m/m), giving rise to a recovery of 65.6%. Circular dichroism spectra indicate no conformational change for cytochrome C after the adsorption and desorption processes, demonstrating the favorable biocompatibility of TeW6 @ZIF-8 composite. In applying practical samples, SDS-PAGE results showed that cytochrome C was successfully isolated and purified by TeW6 @ZIF-8 composite from porcine heart protein extract, which is further identified with LC-MS/MS. Thus, a new strategy for separating and purifying cytochrome C from the porcine heart using TeW6 @ZIF-8 composite as an adsorbent was established.

Reduced CCR6+IL-17A+Treg Cells in Blood and CCR6-Dependent Accumulation of IL-17A+Treg Cells in Lungs of Patients With Allergic Asthma.

Human regulatory T (Treg) cells play a central role in controlling allergic inflammation in the airways. A reduced number of peripheral Treg cells and decreased suppressive function have been previously reported in the pathogenesis of allergic asthma. However, the characteristic role of specific Treg cell subsets and their mechanisms in the pathogenesis of allergic asthma remain unclear. In this study, we examined the proportion of different Treg cell subsets in both healthy subjects and patients with allergic asthma using flow cytometry and single-cell RNA sequencing. The migration function of the cells was compared using cell sorting and Transwell experiments. Furthermore, two allergen-challenged mouse models and a cell transfer experiment were used to examine the role of these Treg subsets. We found that the proportion of CD25+Foxp3+CD127- Treg cells in the peripheral blood of patients with allergic asthma was lower than in those of healthy subjects. Furthermore, the circulating Treg cells expressed lower levels of CCR6 and IL-17 compared with healthy subjects. The chemokine from the airway mucosa, CCL20, was abundantly expressed, and Transwell experiments further proved that this chemokine promoted CCR6+ Treg cell migration in vitro. A mouse model induced by house dust mite (HDM) revealed that the number of CCR6+ Treg cells in the lung tissue increased remarkably. The incidence of allergic asthma may be related to an increase in Treg cells secreting IL-17 in the lung tissue. Recruited CCR6+ Treg cells are likely to differentiate into Th17-like cells under the Th17 environment present in the lungs. IL-17 derived from Th17-like cells could be associated with the pathology of allergic asthma by promoting Th17 responses, thereby favoring HDM-induced asthma exacerbations.

Upregulated Expression of Toll-Like Receptor 7 in Peripheral Blood Basophils of Patients With Allergic Rhinitis.

BACKGROUND: Recently, it has been reported that Toll-like receptor 7 (TLR7) agonists can improve allergic rhinitis (AR) symptoms by up-regulation of Th1 cytokine release and suppression of Th2 cell functions. However, little is known of the expression of TLR7 in basophils of AR. OBJECTIVE: To explore the expression of TLR7 in basophils of AR, and influence of allergens on TLR7 expression. METHODS: The expression levels of TLR7 in basophils of patients with AR were determined by flow cytometry, and the influence of allergens on TLR7 expression was examined by real time (q) PCR. RESULTS: The percentages of TLR7+CCR3+ cells (P < 0.001 and P = 0.011), TLR7+CD123+HLA-DR- cells (P = 0 .016 and P = 0.042) and TLR7+CCR3+CD123+HLA-DR- cells (P = 0.046 and P = 0.035) in blood granulocyte and mononucleated cell populations of the patients with AR were increased, respectively compared with HC subjects. TLR7 MFI on CCR3+ cells (P = 0.050 and P = 0.043), CD123+HLA-DR- cells (P < 0.001 and P = 0.002) and CCR3+CD123+HLA-DR- cells (P < 0.001 and P = 0.003) were enhanced compared with HC subjects. Allergens Der p1 and OVA provoked upregulation of TLR7 expression at both protein and mRNA levels and IL-13 production in KU812 cells. House Dust Mite extract (HDME), Artemisia sieversiana wild allergen extract (ASWE), IL-31, IL-33, IL-37, and TSLP provoked elevation of IL-6 release from KU812 cells following 2 h incubation period. CONCLUSIONS: The percentage of TLR7+ basophils and TLR7 expression intensity in a single basophil are both increased in the blood of patients with AR, indicating that basophils likely contribute to the pathogenesis of AR via TLR7.

Accumulative Deformation Characteristics and Microstructure of Saturated Soft Clay under Cross-River Subway Loading.

The cross-river subway in the Hangzhou Bay area often passes through deep, thick, soft soil at the bottom of the river. At the same time, overlying erosion, siltation, and changes in water levels adversely affect the deformation of the subway, thereby causing hidden dangers to its safe operation. Using two-way dynamic triaxial testing, the effects of cyclic loading of the cross-river subway on the soft clay foundation were investigated for the first time, using simulation methodology as the prime objective of the present study. A strain development curve for the soft clay was obtained as a result. Considering the effects of effective confining pressure (p') and radial cyclic stress ratio (τr), an explicit model of accumulative strain on soft clay under cyclic loading of the cross-river subway was established. The results showed that the accumulative axial strain (εd) was closely related to p' and τr. Under certain conditions, as p' and τr increased, the εd produced by the soil tended to decrease. Furthermore, through non-destructive testing based on nuclear magnetic resonance (NMR), pore distribution and pore size changes in soft clay during cyclic loading were analyzed. It was observed that under the action of the cross-river loading, the large internal soft clay pores were transformed into small pores, which manifested as a significant decrease in the number of large pores and an increase in the proportion of small pores. Lastly, the macroscopic dynamic soil characteristics observed during triaxial testing closely correlated with the microscopic pore size of the soil obtained in the NMR test, which indicated that using pore distribution and pore size changes to describe microscopic changes was a valid method.

Upregulation of the expression of Toll-like receptor 9 in basophils in patients with allergic rhinitis: An enhanced expression by allergens.

It was reported that the expression of Toll-like receptor (TLR) 9 may be related to Th2-type allergic inflammation including allergic rhinitis (AR). However, little is known about the expression of TLR9 in the basophils in AR. In the present study, the expression of TLR9 was examined by flow cytometry analysis, and the expression of TLR9 mRNA in KU812 was determined by quantitative real-time PCR. The results showed that the percentage of TLR9+ CCR3+ cells in blood granulocytes increased by 46% in patients with AR, but not in peripheral blood mononuclear cells (PBMCs). Allergens namely Dermatophagoide allergen extract (DAE) and Platanus pollen allergen extract (PPAE) upregulated the expression of TLR9 in CCR3+ granulocytes by 76% and 84%, respectively. DAE and PPAE also enhanced the proportions of TLR9+ CD123+ HLA-DR- cells and TLR9+ CCR3+ CD123+ HLA-DR- cells in granulocytes and PBMCs of patients with AR. In order to investigate the actions of allergens on basophils, KU812 cells were used. It was observed that all KU812 cells expressed TLR9, and the expression intensity of TLR9 in a single KU812 cell was elevated by CpG. IL-37, IL-31, IL-33, Artemisia sieversiana wild allergen extract (ASWAE), DAE, OVA and Der p 1 induced an increase in the expression of TLR9 mRNA and IL-6 production in KU812 cells. It was shown that the percentage of TLR9-expressing basophils increased in the blood of ovalbumin (OVA)-sensitized mice. In conclusion, an increased expression of TLR9 and the production of IL-6 in basophils implicate that the contribution of basophils to AR is likely via TLR9.

Poly(Acrylic Acid)-Modified MoS2 Nanoparticle-Based Transdermal Delivery of Atenolol.

INTRODUCTION: Hypertension is a major health problem worldwide and is typically treated using oral drugs. However, the frequency of oral administration may result in poor patient compliance, and reduced bioavailability owing to the first-pass effect can also prove problematic. METHODS: In this study, we developed a new transdermal-drug-delivery system (TDDS) for the treatment of hypertension using atenolol (ATE) based on poly(acrylic acid) (PAA)-decorated three-dimensional (3D) flower-like MoS2 nanoparticles (PAA-MoS2 NPs) that respond to NIR laser irradiation. The PAA-modified MoS2 NPs were synthesized and characterized using attenuated total reflection Fourier-transform infrared spectroscopy, X-ray diffraction measurements, scanning electron microscopy, transmission electron microscopy, dynamic light scattering, and the sedimentation equilibrium method. The drug-loading efficiency and photothermal conversion effect were also explored. RESULTS: The results showed that the colloidally stable PAA-MoS2 NPs exhibited a high drug-loading capacity of 54.99% and high photothermal conversion ability. Further, the capacity of the PAA-MoS2 NPs for controlled release was explored using in vitro drug-release and skin-penetration studies. The drug-release percentage was 44.72 ± 1.04%, and skin penetration was enhanced by a factor of 1.85 in the laser-stimulated group. Sustained and controlled release by the developed TDDS were observed with laser stimulation. Moreover, in vivo erythema index analysis verified that the PAA-MoS2 NPs did not cause skin irritation. DISCUSSION: Our findings demonstrate that PAA-MoS2 NPs can be used as a new carrier for transdermal drug delivery for the first time.

Immunological characterization of the American cockroach allergen Per a 9 expressed in baculovirus-infected insect cells.

American cockroach (CR) allergy has been recognized as important IgE-mediated type I hypersensitivity. Per a 9 is an arginine kinase, reacting with IgE in sera of all CR allergic Thai patients. Per a 9 gene was cloned and expressed in eukaryotic systems (baculovirus-infected insect cells). The expressed Per a 9 was purified by Nickel column. The antigenicities were analyzed by ELISA, immunoblot analysis and basophile activation test. The results show that 13 out of 16 (81.3%) sera from American CR patients reacted to Per a 9, confirming that Per a 9 is a major allergen of CR. The IgE reactivity of Per a 9 in the sera from American CR patients was increased 8.3-fold in comparison with the sera from healthy controls. Per a 9 at 1.0 µg/ml induced an approximately up to 5.6-fold increase in CD63 and CCR3 double positive cells when incubating with passively sensitized basophils from by sera from American CR patients.

Expression of toll-like receptors and their regulatory roles in murine cardiac telocytes.

Telocytes, newly discovered in the last decade, are interstitial cells found in numerous organs, with multiple proposed potential biological functions. Toll-like receptors (TLRs) play an important role in innate and adaptive immunity by recognizing pathogen-associated molecular patterns (PAMPs). However, it is still unknown whether telocytes express these innate receptors. We sought to determine the expression and role of TLRs in telocytes. In our study, we primarily detected TLR1-9 expression in telocytes. The proliferation, apoptosis and immunoregulatory activity of telocytes activated with or without TLR ligands were determined. Our results showed that purified telocytes expressed TLR2, TLR3 and TLR5. In particular, telocytes expressed high levels of TLR2 as observed using flow cytometry. When we stimulated telocytes with TLR2 or TLR3 agonists (Pam3CSK4, PolyI:C), iNOS expression was greatly increased after Pam3CSK4 treatment. Additionally, telocyte proliferation was reduced and cell apoptosis was increased after TLR agonist stimulation. A co-culture experiment showed that supernatant from telocytes pretreated with Pam3CSK4 inhibited T cell activation much more than that from untreated telocytes and this effect was mediated by iNOS. Overall, our results demonstrated TLR expression on telocytes for the first time and provided evidence of an immunoregulatory role of telocytes, indicating their clinical potential.

Chinese Society of Allergy Guidelines for Diagnosis and Treatment of Allergic Rhinitis.

Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2-3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of Journal Articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.

Interleukin 29 activates expression of tissue inhibitor of metalloproteinase 1 in macrophages via toll­like receptor 2.

Obesity and diabetes are characterized by low grade chronic inflammation status and insulin resistance in adipose tissue associated with metalloproteinase inhibitor 1 (TIMP1). Interleukin (IL)29, exhibits multiple immune regulatory activities. However, the role of IL29 and its effects on TIMP1 remain to be elucidated. The present study was designed to investigate the effects of IL29 on expression of TIMP1 in macrophages associated with inflammation in adipose tissue. IL29 and high glucose were used to activate Raw264.7 cells and primary macrophages with or without antibody­mediated inhibition of toll like receptor (TLR) 2 and TLR4. TIMP1 was measured in culture media of Raw264.7 cells and primary macrophages by ELISA. IL29 and high glucose increased TIMP1 levels in Raw264.7 cells and primary macrophages. Antibody­mediated inhibition of TLR2 or TLR2 gene knockout decreased TIMP1 levels activated by IL29, however not by high glucose in the medium of Raw264.7 cells and primary macrophages. Furthermore, antibody­mediated inhibition of TLR4 or TLR4 gene knockout decreased TIMP1 levels which were stimulated by high glucose, not by IL29 in the medium of Raw264.7 cells and primary macrophages. The results of the present study indicate that TLR2 is involved in IL29­stimulated TIMP1 expression in Raw264.7 cells and primary macrophages.

Altered expression of IL-18 binding protein and IL-18 receptor in basophils and mast cells of asthma patients.

IL-18 is likely to contribute to asthma. However, little is known regarding the role of IL-18 binding protein (BP) and IL-18 receptor (R) in asthma. Because the action of IL-18 in the body is regulated by IL-18BP and mast cells and basophils are key cell types involved in asthma, we investigated the expression of IL-18, IL-18BP and IL-18R in basophils and mast cells using flow cytometry and a mouse asthma model. We found that among basophils, approximately 53% and 51% were IL-18+ , 85% and 81% were IL-18BP+ basophils, and 19.8% and 8.6% were IL-18R+ in healthy control (HC) and asthmatic blood, respectively. The allergens tested had little effect on the expression of IL-18 and related factors. Only 3.5%, 14.3% and 2.4% of dispersed mast cells expressed IL-18, IL-18BP and IL-18R, respectively, in asthmatic sputum. In a mouse asthma model, OVA-sensitized mice exhibited decreased IL-18BP+ but increased IL-18R+ basophils in their blood. IL-18 increased the number of basophils but eliminated IL-18BP+ basophils in mouse blood. IL-18 increased the number of mast cells and IL-18R+ mast cells in the lung as well as increased the mast cell numbers and IL-18BP+ mast cells in the bronchoalveolar lavage fluid (BALF) of OVA-sensitized mice. Thus, basophils and mast cells may be involved in asthma pathogenesis via an IL-18-associated mechanism.

Role of IL-18 in atopic asthma is determined by balance of IL-18/IL-18BP/IL-18R.

It is recognized that IL-18 is related to development of asthma, but role of IL-18 in asthma remains controversial and confusing. This is largely due to lack of information on expression of IL-18 binding protein (BP) and IL-18 receptor (R) in asthma. In this study, we found that plasma levels of IL-18 and IL-18BP were elevated in asthma. The ratio between plasma concentrations of IL-18 and IL-18BP was 1:12.8 in asthma patients. We demonstrated that 13-fold more monocytes, 17.5-fold more neutrophils and 4.1-fold more B cells express IL-18BP than IL-18 in asthmatic blood, suggesting that there is excessive amount of IL-18BP to abolish actions of IL-18 in asthma. We also discovered that more IL-18R+ monocytes, neutrophils and B cells are located in asthmatic blood. Once injected, IL-18 eliminated IL-18R+ monocytes in blood, but up-regulated expression of IL-18R in lung macrophages of OVA-sensitized mice. Our data clearly indicate that the role of IL-18 in asthma is very likely to be determined by balance of IL-18/IL-18BP/IL-18R expression in inflammatory cells. Therefore, IL-18R blocking or IL-18BP activity enhancing therapies may be useful for treatment of asthma.