In recent years, the role of the monocyte macrophage system and Toll like receptor 9 (TLR9) in the pathogenesis of allergic diseases has attracted attention, but little is known about the relationship between changes in TLR9 expression on monocytes and macrophages and allergic rhinitis (AR) and allergic asthma (AA). This article uses flow cytometry to analyze the expression changes of TLR9+ monocytes in peripheral blood of patients with AR, AA, and allergic rhinitis combined with asthma (ARA). And the AR and AA models of wild-type (WT) and Fc epsilon RI gene knockout (Fc epsilon RI KO) mice were used to investigate the expression changes of TLR9 in monocytes and macrophages in lung tissue of mice after allergen stimulation. It was found that the proportion of TLR9 + cells in the peripheral blood mononuclear cell population of AR, AA, and ARA patients was significantly increased, indicating an increase in MFI expression representing the number of TLR9 molecules expressed in each mononuclear cell. Artemisia selengensis allergen extract (ASWE), house dust mite allergen extract (HDME), wutong pollen allergen extract (PPE) stimulate to increase the proportion of TLR9+ monocytes in the blood of AR and AA model mice, upregulate the expression level of TLR9 MFI in monocytes, and make the expression level of TLR9 in macrophages in lung tissue higher than that of control mice. In AR and AA mouse models with high-affinity IgE receptor (Fc epsilon RI) KO, ASWE, especially HDME, can also induce upregulation of TLR9 + cell percentage and TLR9 MFI in monocytes. Based on the significant reduction in the proportion of TLR9+ cells and TLR9 MFI expression levels in AR and AA mice compared to wild-type (WT) mice, it can be concluded that the increase in the proportion of blood monocytes induced by allergen extract and the upregulation of TLR9 expression levels in monocytes are achieved through both Fc epsilon RI dependent and non-dependent pathways. Similarly, compared with WT mice, the AA model mice of Fc epsilon RI KO showed a significant increase in the proportion of lung tissue macrophages induced by allergen extract and a significant decrease in the upregulation of TLR9 expression level. In summary, monocytes may participate in the pathophysiological processes of AR, AA, and ARA through upregulation of TLR9 expression. The upregulation of TLR9 expression in lung tissue macrophages further suggests that monocytes and macrophages may form a new inflammatory signal amplification loop in local tissues through high expression of TLR9 under allergen stimulation, providing new ideas for understanding the pathogenesis of allergic airway diseases.

Altered basophil identification markers have been discovered to associate with allergic asthma (AA) recently. However, little is known of the expressions of basophil markers in blood granulocytes. We therefore parallel tested them in peripheral blood mononuclear cell (PBMC) and granulocyte populations of patients with AA and combined allergic rhinitis and asthma (ARA) by flow cytometry and assessed their diagnostic performance and examined plasma levels of EDN, LTC4 and PGD2. We found CCR3+ cell numbers increased markedly in granulocytes of AA and ARA patients. Almost all isolated CCR3+ PBMC were basophils, but CCR3+ granulocytes contained up to 96.0% eosinophils and 12.8% basophils. The numbers of CD63, CD203c and FcεRIα expressing CCR3+ granulocytes increased markedly, and the mean fluorescent intensity of CD203c, FcεRIα and CD63 expression on CCR3+ granulocytes and PBMC enhanced consistently in AA and ARA patients. The area under the curve achieved the highest values for CD203c+CCR3+ granulocyte numbers in AA and ARA patients. Plasma levels of EDN, LTC4 and PGD2 in AA and ARA patients increased and correlated well with CCR3+ granulocyte numbers. In conclusion, discovering basophils in peripheral blood granulocytes provides greater scope for the clinical use of basophil tests.

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.

Background: Panax notoginseng (PNE) is a prominent traditional Chinese medicine with extensive beneficial effects on the immune system. However, the precise mechanism of PNE in treating inflammatory bowel disease (IBD) remains unclear. Methods: We first used an extensive metabolomics approach utilizing UPLC-ESI-Q TRAP-MS/MS to identify the metabolite components of PNE aqueous extract. Moreover, the mechanism of PNE in treating IBD was investigated through in silico analysis including RNA-seq analysis, Network pharmacology and Molecular docking. Then a Drosophila toxin-induced intestinal inflammation model was employed to investigate further. Results: A total of 1,543 metabolites of PNE aqueous extract were characterized using UPLC-ESI-Q TRAP-MS/MS. In silico analyses showed that 97 IBD hub targets were targeted by 21 PNE ingredients. Kyoto Encyclopedia of Genes and Genomes results indicated that PNE may play an anti-IBD role through the Mitogen-activated protein kinase (MAPK) signaling pathway and other immune-related signaling pathways. Moreover, 11 top hits compounds of PNE show a good affinity binding to IBD targets. The experimental results demonstrated that PNE can effectively improve the survival rate of adult Drosophila while also inhibit the excessive proliferation and differentiation of intestinal stem cells induced by sodium dodecyl sulfate. Furthermore, PNE notably lower the epithelial cell mortality, the accumulation of reactive oxygen species and the activation of oxidative stress-associated jun-Nterminal kinase (JNK) pathway. Conclusion: Our data suggests that PNE aqueous extract has a significant protective impact on the intestinal homeostasis of Drosophila. These findings establish a basis for utilizing PNE in clinical investigations and managing

目的 检测特应性皮炎(atopic dermatitis,AD)患者外周血调节性T细胞亚群及相应细胞因子的表达,探讨其与内源性AD和外源性AD诊断及临床分型的意义.方法 选择36例AD患者(内源性AD组、外源性AD组各18例)和15例健康人(对照组),采用流式细胞仪检测各组患者外周血中CD4+CD25+Foxp3+Treg细胞、CD8+Foxp3+Treg细胞和IL-17+Foxp3+Treg细胞及其细胞因子IL-10的表达情况.结果 与对照组相比较,内源性AD组和外源性AD组患者外周血CD4+CD25+Foxp3+Treg细胞、CD8+Foxp3+Treg细胞、IL-10+Foxp3+Treg细胞百分比均明显降低(P<0.05),内源性AD组IL-17+Foxp3+Treg细胞百分比明显增高(P<0.05),外源性AD组IL-17+Foxp3+Treg细胞百分比差异无统计学意义(P>0.05).内源性AD组和外源性AD组CD4+CD25+Foxp3+Treg细胞、CD8+Foxp3+Treg细胞及IL-10+Foxp3+Treg细胞百分比差异均无统计学意义(P>0.05).结论 AD患者外周血CD4+CD25+Foxp3+Treg细胞、CD8+Foxp3+Treg细胞及IL-10+Foxp3+Treg细胞比例降低,可能导致了AD疾病的发生、发展,并且在内源性AD和外源性AD中都有参与,IL-17+Foxp3+Treg细胞在内源性AD的发病机制中占据主导地位.

目的 探讨特应性皮炎(AD)患者外周血中嗜碱性粒细胞CD63、CD203c和FceRI的表达及其临床意义.方法 收集36例AD患者(内源性AD组、外源性AD组各18例)和20例健康人(HC)的外周血,流式细胞仪检测并比较AD患者(tAD组)、内源性AD患者(iAD组)和外源性AD患者(eAD组)嗜碱性粒细胞富集群中CD63、CD203c和FceRI的表达情况.结果 相比于健康对照组,AD患者粒细胞群中嗜碱性粒细胞平均荧光强度(mean fluorescence intensity,MFI)显著增加(P＜0.01),粒细胞群中CD63+嗜碱性粒细胞MFI升高(P＜0.05),单个核细胞群中CD63+嗜碱性粒细胞比例较健康对照组显著增加(P＜0.01),粒细胞群CD203c+嗜碱性粒细胞MFI升高(P＜0.05),粒细胞群CD203c+嗜碱性粒细胞比例显著升高(P＜0.01),单个核细胞群中CD203c+嗜碱性粒细胞比例升高(P＜0.05),粒细胞群中FcεRI+嗜碱性粒细胞比例显著升高(P＜0.01).内源性AD与外源性AD在嗜碱性粒细胞及其表面CD63、CD203c和FcεRI的表达上差异均无统计学意义(P＞0.05).结论 AD患者嗜碱性粒细胞表达CD63、CD203c和FceRI增强,检测嗜碱性粒细胞及其相关分子对诊断AD具有意义.

Interleukin (IL)‐18 is a potentially important molecule in allergic rhinitis (AR). However, expressions of IL‐18, IL‐18 binding protein isoform a (IL‐18BPa) and IL‐18 receptor alpha (IL‐18Rα) in AR blood monocytes remain obscure. We, therefore, investigated IL‐18, IL‐18BPa and IL‐18Rα expressions in monocytes using flow cytometry, murine AR model and quantitative real‐time polymerase chain reaction (PCR). The results showed that the numbers of IL‐18 + monocytes increased, whereas IL‐18BPa + monocytes decreased in the peripheral blood of AR patients. It was also observed that Platanus pollen extract provoked elevated expressions of IL‐18 and IL‐18Rα in the monocytes of AR patients. House dust mite extract, Artemisia sieversiana wild extract and Platanus pollen extract enhanced IL‐18Rα protein and mRNA expression in the isolated primary monocytes from AR patients. Using ELISA kits, we observed that the levels of total IL‐18 and free IL‐18 in the plasma of perennial AR (pAR) and seasonal AR (sAR) patients were elevated, and the molar concentration ratio of free IL‐18BPa/free IL‐18 was 16.5 for healthy control subjects and 9.7 for patients with sAR, indicating that IL‐18 likely plays a role in sAR. In the murine AR model, the number of IL‐18Rα + monocytes increased in the blood, and the number of IL‐18Rα + macrophages increased in the nasal lavage fluid of WT mice. In conclusion, IL‐18 may serve as a causative factor for AR.

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. Fc epsilon RI-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.

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.

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.

目的:探讨 Th2 细胞趋化因子同源分子(chemoattractant receptor homologous molecule expressed on Th2 cells,CRTh2)、半胱氨酸白三烯受体 1(cysteinyl leukotriene 1 receptor,CysLT1R)及组胺 1 型受体(histamine 1 receptor,H1R)在支气管哮喘(bronchial asthma,BA)患者外周血CD3+CD56+自然杀伤T细胞(natural killer T cells,NKT)中的表达情况及临床意义.方法:收集2020年9月至2021年9月锦州医科大学附属医院BA患者(BA组,n=40)和健康对照组(healthy control,HC组,n=38)的外周血,流式细胞仪检测BA组和HC组外周血中CRTh2、CysLT1R及H1R在CD3+CD56+标记的NKT细胞上的表达,同时对其与血清总 IgE(total immunoglobulin E,TIgE)、呼出气一氧化氮(fractional exhaled nitric oxide,FeNO)及肺功能第一秒用力呼气量/用力肺活量％(forced expiratory volume in the first second/forced vital capacity％,FEV1/FVC％)的相关性分析.结果:BA 组[22.50(15.78,29.20)]外周血 CD3+CD56+NKT 细胞百分比较 HC 组[27.90(21.50,32.45)]降低(P=0.024);进一步比较 BA 组[70.05(63.20,78.45)]CD3+淋巴细胞百分比低于 HC 组[73.70(69.88,79.38)](P=0.035).BA 组[0.08(0.05,0.17)]CysLT1R 百分比较 HC 组[0.16(0.10,0.34)]降低(P=0.004);BA 组 CRTH2、H1R 百分比[1.12(0.53,1.88);5.21(1.41,10.10)]与 HC 组[1.17(0.60,4.25);4.61(1.42,8.53)]比较,无统计学差异(P=0.234;P=0.697).进一步对比 CRTh2、H1R 的平均荧光强度(mean fluorescence intensity,MFI),BA 组[925.52(623.50,1 279.25)]CRTh2 MFI 较 HC 组[1 345.50(891.00,1 977.75)]降低(P=0.002);BA 组[769(657.50,975.00)]H1R MFI 与 HC 组[835.00(560.00,959.25)]无统计学差异(P=0.834).CD3+CD56+NKT 细胞与血清 TIgE、FeNO 无明显相关性(r=-0.052、r=0.014),与 FEV1/FVC％水平呈正相关(r=0.402,P=0.01).CRTH2(r=0.166、r=-0.040)、CysLT1R(r=0.114、r=-0.150)及H1R(r=0.103、r=-0.096)与血清TIgE、FeNO无明显相关性.结论:CD3+CD56+NKT细胞可能在支气管哮喘发病过程中起重要作用,H1R在CD3+CD56+NKT细胞上可能不表达,CRTh2与CysLT1R在BA患者CD3+CD56+NKT细胞上表达减少可能会为临床精准诊疗提供依据.

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, TLR7+CD123+HLA-DR- cells and TLR7+CCR3+CD123+HLA-DR- cells in blood granulocyte and mononucleated cell populations of the patients with AR were increased, respectively compared with HC subjects. TLR7 MFI on CCR3+ cells, CD123+HLA-DR- cells and CCR3+CD123+HLA-DR- cells were enhanced. 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. Trial Registration: Trial registry: Chinese clinical trial; registration number: ChiCTR-BOC-16010279.

目的 探讨血液嗜碱性粒细胞CD16和CD32在哮喘-慢性阻塞性肺疾病重叠(ACO)、慢性阻塞性肺疾病(COPD)中的表达及其临床意义.方法 选取2019年12月-2020年12月就诊于锦州医科大学附属第三医院的COPD、ACO患者及招募的健康志愿者作为研究对象,分别设为COPD组、ACO组及健康对照组.收集研究对象的性别、年龄、病史、发病年龄、体重指数(BMI)及吸烟史等一般资料并采集外周静脉血,采用流式细胞仪检测嗜碱性粒细胞富集群中CD16和CD32的表达情况,ELISA法检测血浆中肿瘤坏死因子-α(TNF-α)的表达水平.结果 与健康对照组相比,ACO组、COPD组外周血单个核细胞群中CD16+CCR3+细胞、CD16+CD123+HLA-DR–细胞、CD16+CCR3+CD123+HLA-DR–细胞比例增高(P<0.05),COPD组单个核细胞群中CD32+CD123+HLA-DR–细胞、CD32+CCR3+CD123+HLA-DR–细胞比例增高(P=0.003,P=0.030),ACO组粒细胞群中CD32+CD123+HLA-DR–细胞、CD32+CCR3+CD123+HLA-DR–细胞比例降低(P=0.019,P=0.031).ACO组粒细胞群中CCR3+细胞和CCR3+CD123+HLA-DR–细胞的CD32平均荧光强度(MFI)较健康对照组降低(P=0.006,P=0.047).ACO组血浆TNF-α表达水平较健康对照组升高,差异有统计学意义(P=0.036).结论 嗜碱性粒细胞源CD16和CD32可能在ACO、COPD发病过程中发挥重要作用,TNF-α可能参与了ACO的发病,为后续ACO和COPD的临床治疗提供了新的靶点.

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.

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.

Objective:To optimize the BALB/c mouse rhinitis model sensitized by Artemisia annua pollen allergen, and explore the humoral and cellular immune indicators that can be used for the evaluation of allergic reactions. Methods:Using BALB/c mice as experimental animals, using Artemisia annua pollen allergen extract as sensitizing protein, through different content of the main allergen Art a1 and different sensitization times, different immunization programs were set to immunize mice subcutaneously, One week and five weeks after the last immunization, Artemisia annua pollen allergen extract containing 50 μg/ml and 500 μg/ml Art a1 was used for nasal stimulation, once a day, for 1 week each time.Observe the allergic reaction of mice, detect the pathological changes of nasal tissues, determine the levels and dynamic changes of antigen-specific IgE, IgG1, IgG2a and other antibodies in the serum of each group of mice. and detect the changes in the number of antigen-specific IL-4, IL-5, IL-2, IFN-γ and other lymphocytes in the spleen of mice. Results:Sensitized mice showed obvious scratching and sneezing reactions after being stimulated by antigen; obvious allergic inflammation appeared in nasal tissue; The increase in serum level of Artemisia annua pollen-specific IgE antibody was significantly correlated with the challenge antigen; The antigen-specific IL-4 lymphocytes in the spleen of the sensitized mice were significantly increased, but the IFN-γ-specific lymphocytes did not change significantly. Conclusions:The successful establishment of a mouse model of Artemisia annua pollen allergen allergy is the first domestic use of ELISPOT technology to detect an increase in the number of antigen-specific IL-4 lymphocytes in Artemisia annua allergy mice, laying a foundation for the subsequent evaluation of the efficacy of preparations for desensitization treatment basis.

Background: ACO has been characterized as a kind of clinical disease with overlap symptoms of asthma and COPD. However, little is known of the role of eosinophils and corresponding cytokines and chemokines in ACO patients with different treatment responses. Methods: To evaluate factors which associates with different treatment responses in patients with ACO. In the present study, we investigated the eosinophils proportion of peripheral blood from ACO patients with acute exacerbation (AE) after treatment, ACO patients with clinical response (CR) after treatment, and healthy volunteers (HV) by using flow cytometry analysis. The plasma levels of corresponding cytokines and chemokines from the three groups were evaluated by ELISA. Results: The results showed that ACO patients that had acute exacerbation have relatively lower eosinophils proportions compared to healthy volunteers but have higher eosinophils inflammation compared with patients with clinical response. The percentage of NK1R+ expression population eosinophils was also decreased. Further analysis revealed ACO patients that had acute exacerbation also have relatively higher plasma levels of cytokines and chemokines compared to patients with clinical response after treatments and healthy volunteers. Conclusion: ACO patients from AE group have relatively lower eosinophil proportion and NK1R+ expression population eosinophil proportion, but higher plasma levels of cytokines and chemokines. Inhibitors of cytokines and chemokines are likely useful agents for treatment of ACO patients which had acute exacerbations.

Background: Interleukin (IL)-18 is emerging as an attractive participant in allergic rhinitis (AR). However, correlation of IL-18 with IL-18 binding protein (BP) in plasma, and expression of IL-18, IL-18BP and IL-18 receptor (R) in AR blood monocytes remain obscure. We therefore investigated IL-18, IL-18BP and IL-18R expression in monocytes using flow cytometric analysis, murine AR model, and quantitative real-time PCR in the present study. Results: Plasma IL-18 and IL-1β in AR patients was higher than those in healthy control subjects. Free (f)IL-18 had a high correlation with IL-18BP, IL-1β and TNF-α in AR plasma. Proportion of IL-18 + monocytes was increased, whereas IL-18BP + monocytes were decreased in blood of patients with AR. It was found that Platanus pollen allergen extract provoked the elevated expression of IL-18 and IL-18R in AR blood monocytes. Dermatophagoides pteronyssinus , Artemisia sieversiana wild and Platanus pollen allergen extracts enhanced IL-18R protein and mRNA expression in isolated primary monocytes from AR patients. Moreover, numbers of macrophages and IL-18R + macrophages in nasal lavage fluid (NLF) were increased, and levels of IL-18 in both plasma and NLF were elevated in AR mice. Conclusions: These suggest that IL-18 is likely to participate in the development of AR as a causative factor; therefore, it could be a therapeutic target for AR.

In recent years, it's recognized that immunity is controlled by regulatory T cell (Treg). Since essential pathophysiological vagaries of allergy are primarily caused by hyper responsiveness of system to allergens that obtains after birth, Tregs probable play important roles within the pathogenesis of allergy, predominantly in the sensitization phase. The aim of this study is to aim to classify subtypes of Tregs and summarize their roles in allergy. Tregs must contain natural Tregs (nTreg) counting inducible costimulator (ICOS) (+) Tregs, CD8(+) Tregs and IL-17-producing Tregs, interleukin (IL)-10-producing type 1 Tregs (Tr1 cells), inducible/adaptive Tregs (iTreg). These cells share some conjoint features as well as appearance of Foxp3, and secretion of inhibitory cytokine IL-10 and/or TGF-β. Moreover, it's obvious that Tregs likely pay to allergic conditions like airway inflammation and dermatitis, and plays a significant role within the conduct of allergy complete their actions on conquest of effector T cells and self-consciousness of initiation of mast cells and basophils. Modulation of functions of Tregs may provide a completely unique strategy to stop and treat allergic diseases.