The Effects of Microbial Materials Adhered to Asian Sand Dust on Allergic Lung Inflammation

Asian sand dust (ASD) containing microbiological materials, sulfate (SO₄²), and nitrate (NO₃⁻) derived from air pollutants in East China, reportedly cause adverse respiratory health effects. ASD aggravates ovalbumin (OVA)-associated experimental lung eosinophilia. In this study, the toxic materials adsorbed onto ASD were excluded by heat treatment at 360°C for 30 min. The effects of nonheated ASD or heated ASD (H-ASD) toward the allergic lung inflammation were compared in murine lungs. ICR mice were administered intratracheally with normal saline (control), H-ASD, ASD, OVA, OVA + H-ASD, and OVA + ASD, four times at 2-week intervals. ASD only increased neutrophils in bronchoalveolar lavage fluids (BALFs) along with pro-inflammatory mediators, such as keratinocyte chemoattractant (KC). H-ASD and ASD enhanced eosinophil recruitment induced by OVA in the alveoli and in the submucosa of the airway, which has a goblet cell proliferation in the bronchial epithelium. The two ASDs synergistically increased interleukin-5 (IL-5), monocyte chemotactic protein-3 (MCP-3), and eotaxin, which were associated with OVA, in BALF. The enhancing effects were much greater in ASD than in H-ASD. The two ASDs induced the adjuvant effects to specific IgE and IgG1 production by OVA. In the in vitro study using RAW264.7 cells, ASD increased the expression of Toll-like receptor 2 (TLR 2) mRNA but not TLR4 mRNA. H-ASD caused no expression of either TLR mRNA. These results suggest that the aggravated lung eosinophilia by ASD may be due to activation of Th2-associated immune response via the activation of TLR2 by microbial components adhered to ASD.     

重组粉尘螨Ⅱ类变应原致敏小鼠肺部变应性炎症模型的建立

目的探讨以重组粉尘螨Ⅱ类变应原(rDerf2)建立小鼠肺部变应性炎症模型的方法。方法将30只6～8周龄雌性SPF级BALB/c小鼠,随机分为PBS组(阴性对照组)、卵清蛋白(OVA)组(阳性对照组)和rDerf2组(实验组)。实验组分别于第0、7、14天用rDerf2对小鼠腹腔进行注射致敏;再于第21天开始,连续7d进行雾化激发;对照组分别用PBS和OVA代替。最后一次雾化激发后24h内处死动物,观察肺组织病理变化、支气管肺泡灌洗液(BLAF)中的白细胞计数及分类,用ELISA测定BLAF和脾细胞培养上清中白细胞介素2(IL-2)、IL-4、IL-17和IFN-γ的含量及血清中IgG1、IgE抗体水平变化。结果实验组和阳性对照组BALB/c小鼠肺组织呈现明显的炎症性病理改变;rDerf2组的BALF中白细胞总数〔(17.39±1.03)×106/L〕及嗜酸性粒细胞(EOS)〔(1.61±0.03)×106/L〕计数,均明显高于PBS组(P<0.01);BALF中IL-4〔(78.92±9.06)pg/ml〕、IL-17〔(201.63±31.26)pg/ml〕与PBS组比较呈明显的高水平表达(P<0.01);而IL-2〔(8.29±1.27)pg/ml〕和IFN-γ〔(51.04±15.85)pg/ml〕的含量则显著下降(P<0.01);上述指标在脾细胞培养上清中出现类似的变化。rDerf2组血清中IgE〔(37.63±6.57)IU/ml〕、IgG1〔(16.68±2.90)μg/ml〕的含量表现为Th2型反应增强趋势;但OVA组和rDerf2组间所有指标差异均无统计学意义(P>0.05)。结论用rDerf2腹腔注射致敏后再雾化激发的方式能够成功构建小鼠肺部变应性炎症模型。     

支气管哮喘气道IL4和IL12动态变化及对气道炎症反应的影响

目的研究支气管哮喘(哮喘)﹚炎症促进因子白细胞介素4(IL-4)和炎症抑制因子(IL-12)的变化趋势,相互关系及对气道炎症反应的影响.方法用卵蛋白(OVA)﹚敏原制备wistar大鼠哮喘模型,在激发后1、8、24、48、96h及7、14d行支气管肺泡灌洗(BAL),测定支气管肺泡灌洗液(BALF)中炎症细胞总数与嗜酸细胞数(EOS);酶联免疫吸附法(ELISA)测定BALF中IL-4、IL-12的浓度.结果 OVA激发后,气道出现明显的炎症反应.EOS计数1h上升,8h后迅速上升,24h达到高峰期,96h后开始回落,7～14d回落到略高于对照组;炎症细胞总数1h上升,8h后迅速上升,24～96h处于高峰期,7d回落到接近8h水平,14d接近基础水平.BALF中,IL-4在1h上升,8～96h处于高峰期,7～14d回落到基础水平;IL-12在1h上升,8处于低水平,24h上升,48～96h处于高峰期,7～14d回落到基础水平;EOS与IL-4有正相关性(r=0.74,P＜0.001),与IL-12无相关性(r=-0.13,P＞0.05),与IL-4/IL-12有密切相关性(r=0.85,P＜0.001).结论哮喘气道中存在IL-4/IL-12比值失衡,IL-4/IL-12比值对哮喘气道炎症反应起重要的调节作用.     

Liposomal retinoic acids modulate asthma manifestations in mice

Signaling of all-trans retinoic acid (ATRA) through nuclear retinoid acid (RA) receptors regulates several biological functions in airway epithelial cells, eosinophils, and immune cells, yet its impact on different in vivo aspects of pulmonary allergic reaction remains elusive. We compared the effect of a treatment with liposomally encapsulated ATRA (Lipo-ATRA) in a mouse model of ovalbumin (OVA)-induced T helper (Th) 2-type responses and airway remodeling. Daily intraperitoneal injections of 10 mg/kg Lipo-ATRA, at the time of each of the 2 systemic sensitizing injections, increased OVA-induced Immunoglobulin E synthesis, bronchoalveolar lavage (BAL) eosinophilia, and accumulation of IL-5, transforming-growth factor beta1, fibronectin, eotaxin/chemokine (C-C motif) ligand 11 (eotaxin/CCL11) and regulated upon activation, normal T expressed and secreted chemokine (C-C motif) ligand 5. In contrast, Lipo-ATRA, administered during each of the 4 intranasal OVA challenges, did not affect these variables. Regardless of the treatment regimen, Lipo-ATRA augmented mucin levels in BAL fluid and reduced lung total collagen content. In vitro incubation of mouse splenocytes or purified spleen cluster of differentiation (CD) 4-positive T lymphocytes, with ATRA, increased, respectively, OVA- and anti-CD 3 antibody-induced IL-4 and IL-5 production and inhibited IFNgamma release. These findings demonstrate that, when given during systemic sensitization, Lipo-ATRA exacerbates allergic immune and inflammatory responses, most likely by promoting Th2 development.     

内毒素在小鼠过敏性哮喘模型所起作用的实验研究

目的建立小鼠过敏性哮喘模型,在过敏原的致敏和激发阶段给予不同剂量内毒素(即脂多糖LPS),探讨内毒素在哮喘模型中的作用机制。方法建立BALB/c小鼠哮喘模型,设立实验组和对照组开展后续实验。实验组包括:鸡卵清白蛋白(OVA)致敏,OVA激发组;OVA致敏,LPS激发组;OVA致敏,OVA+LPS联合激发组;LPS暴露于OVA致敏前,OVA激发组;对照组包括试剂对照组和健康对照组。各实验组的LPS剂量均分为高、中、低三个剂量组。通过测定小鼠气道阻力和肺顺应性来反映其肺功能改变;流式细胞术(FCM)测定FITC、PE阳性细胞数,表征CD4+CD2+5调节性T淋巴细胞(以下简称Treg细胞)的相对比例;荧光实时聚合酶链反应定量测定肺脏和脾脏Foxp3mRNA表达的水平;ELISA法测定血浆总IgE和OVA特异性IgE水平,抗体夹心ELISA法测定支气管肺泡灌洗液(BALF)中Th2细胞因子(IL-4和IL-5)的水平;计数BALF中白细胞总数和嗜酸性粒细胞等各类白细胞所占百分比;常规病理切片观察肺组织改变情况。结果结果表明,与哮喘模型组小鼠相比,在LPS各处理组中,总体上能诱导Treg细胞的产生,改善肺功能,降低IgE滴度,降低Th2介导的免疫反应,同时诱导肺部炎症。尤其是在致敏阶段腹腔注射内毒素可以缓解由过敏原诱导的气道炎症和改善肺功能,诱导肺组织和脾组织Foxp3mRNA表达的增加。结论LPS抑制了Th2介导的过敏反应,可能与Foxp3表达量的增加相关,后者诱导Treg细胞增殖,对哮喘症状起到缓解作用。     

丹参注射液联合地塞米松对哮喘大鼠CD4+CD25+调节性T细胞的影响

目的 探讨丹参注射液联合地塞米松(DXM)抑制哮喘气道炎症的免疫学机制.方法 50只Wistar大鼠随机分成正常对照(NC)组、哮喘组、丹参组、DXM组、联合用药组,计数支气管肺泡灌洗液(BALF)中细胞总数并分类,HE染色行肺组织病理学检查,流式细胞仪检测外周血单个核细胞(PBMCs)中CD4+CD25+调节性T细胞(CD4+CD25+Treg)比例,ELISA检测BALF中IL-4、IL-5含量.结果 与哮喘组比较,药物干预组细胞总数、中性粒细胞(Neu)、淋巴细胞(Lym)、嗜酸性粒细胞(Eos)百分率下降(P<0.05,P<0.01),联合用药组下降程度大于丹参组、DXM组(P<0.05).哮喘组呈显著炎症变化,丹参组呈中度炎症变化,DXM组呈轻度炎症变化,联合用药组无炎症改变.与哮喘组比较,药物干预组CD4+CD25+Treg/CD4+T升高(P<0.05),IL4、IL-5含量下降(P<0.05),联合用药组CD4+CD25+ Tree/CD4+T升高程度和IL-4、IL-5下降程度大于丹参组和DXM组(P<0.05).结论 丹参注射液具有抑制哮喘大鼠气道炎症的作用,和DXM联合应用后,抑制作用更加明显,该作用可能和促进CD4+CD25+Treg产生,进而抑制IL-4、IL-5产生,纠正Th1/Th2失衡,最终减轻气道炎症有关.     

装饰材料挥发物吸入染毒对大鼠IL-4、IL-5含量及ICAM-1 mRNA表达的影响

[目的]探讨挥发性有机化合物的吸入和呼吸道变态反应之间的关系，为评价装修后室内空气污染提供毒理学依据。[方法]用装饰材料挥发物对雄性SD大鼠静式吸入染毒30d，测定支气管肺泡灌洗液(BALF)中白细胞介素(IL-4、IL-5)含量和肺组织细胞间黏附分子-1(ICAM-1)mRNA表达。[结果]在中浓度染毒组(甲醛浓度为0．81mg／m^3、总挥发性有机化合物(TVOC)浓度为84．88mg／m^3)，BALF中IL-4含量平均为12．95pg／ml，与对照组相比显著升高；在高浓度染毒组(甲醛浓度为1．50mg／m^3、TVOC浓度为299．03mg／m^3)，BALF中IL-4含量平均为13．23pg／ml，IL-5含量平均为23．21pg／ml，均显著高于对照组。中高浓度染毒组大鼠肺组织ICAM-1 mRAN表达显著增强，病理学检查发现肺组织中嗜酸性粒细胞浸润尤为明显。[结论]室内装修可能与呼吸道变态反应性疾病之间有一定联系。     

Therapeutic efficacy of an E coli strain carrying an ovalbumin allergenic peptide as a fused protein to OMPC in a murine model of allergic airway inflammation

An Escherichia coli strain expressing the ovalbumin (OVA) 323-329 allergenic peptide on the bacterial surface was evaluated for its ability to reduce the lung inflammatory response in mice allergic to OVA. BALB/c mice were rendered allergic by means of two intraperitoneal injections of OVA suspended in alum 5 days apart, and one intratracheal boost 1 week later. The mice were then treated with two intranasal, 1 week apart, doses of 4x10(9) E. coli-UH302 transformed with plasmids pST13 or pST13-OVA(323-339), which bear the OmpC porin from Salmonella enterica serovar Typhi or the OmpC with the OVA allergenic 323-339 amino acid sequence inserted in the external loop 5. The allergic inflammatory reaction was evaluated on day 31, finding that mice treated with E. coli-UH302-pST13-OVA reduced four to seven times perivascular and peribronchial infiltrates, mucus production, goblet cell hyperplasia and eosinophils when compared with mice treated with E. coli-UH302-pST13 or saline solution. These results were consistent with a significant decrease of IL-5 mRNA and induction of IFN-gamma mRNA in cells from bronchio-alveolar lavages (BAL). Specific serum IgE anti-OVA was also reduced, although the decrease did not reach statistical significance. These results demonstrate that the bacterial live vector bearing an allergenic peptide successfully moderated two important components of allergy, pulmonary inflammation and mucus overproduction.     

Immunological determinants in a murine model of toluene diisocyanate-induced asthma

Objectives

Diisocyanates (DIC) are highly reactive, low-molecular-weight chemicals which are the leading cause of occupational asthma (OA). The aim of the study was to analyze certain aspects of the pathogenesis of allergic inflammation in the airways induced by toluene diisocyanate (TDI) in an experimental model in mice.

Materials and Methods

The experiment was carried out on 50 female BALB/cJ/Han/IMP mice, which were exposed by inhalation (intranasal and in the inhalation chamber) to toluene diisocyanate (2,4-TDI). After the experiment, the bronchoalveolar lavage fluid (BALF) was collected from the animals, and the composition of the induced inflammatory cells, and the concentrations of certain cytokines (IL-4, IL-5, TNF-α) were evaluated.

Results

The total number of cells in BALF of the examined group of mice was significantly higher compared to the control mice. There was also a significant increase in neutrophils and eosinophils in the study group compared to the controls. The number of lymphocytes and macrophages did not differ significantly between the two groups. A statistically significant increase in the level of TNF-α was shown to occur in the group exposed to toluene diisocyanate in comparison to the control group. The concentration of IL-4 increased in the study group, compared to the control one, but the differences did not reach the level of significance, p > 0.05. Such difference was not observed for IL-5.

Conclusions

We developed a murine model of TDI-induced asthma which caused the influx of inflammatory cells like eosinophils and neutrophils in the bronchoalveolar lavage fluid (BALF) in the TDI-treated mice. The increase of the concentration of some proinflammatory cytokines (TNF-α, IL-4) in BALF from the exposed mice was also observed.     

CD4^＋CD25^＋调节性T细胞对哮喘大鼠气道炎症的影响

目的观察CD4^＋CD25^＋调节性T细胞（CD4^＋CD25^＋Treg）对哮喘大鼠气道炎症的影响。方法将卵白蛋白（OVA）免疫耐受大鼠CD4^＋CD25^＋Treg细胞过继转移给哮喘大鼠,然后观察支气管肺泡灌洗液（BALF）中细胞计数及分类,ELISA检测BALF中IL-4、IL-5和IFN-1及血清OVA特异性IgE含量,HE染色观察肺组织的病理改变。结果与哮喘组比较,过继转移CD4^＋CD25^＋Treg细胞后哮喘大鼠BALF中细胞总数、中性粒细胞和淋巴细胞百分率降低（P〈0．05）,嗜酸性粒细胞（Eos）百分率明显降低（P〈0．01）;BALF中IL4和IL-5含量明显降低,IFN-1含量明显升高,血清OVA特异性IgE含量明显降低（P〈0．05）;气道炎症明显减轻。结论过继转移OVA免疫耐受大鼠CD4^＋CD25^＋Treg细胞可以明显抑制哮喘的慢性气道炎症。     

互隔交链孢霉诱导小鼠气道变态反应性炎症动物模型的构建

目的研究使用互隔交链孢霉孢子构建C57BL/6小鼠肺部变态反应性炎症模型的方法。方法模型组、阳性对照组和阴性对照组各12只小鼠。模型组以互隔交链孢霉孢子悬液致敏和激发小鼠,分别于第0、5、10天腹腔注射霉菌孢子悬液,第14天开始连续4天鼻滴霉菌孢子悬液。阳性对照组以鸡卵蛋白(OVA)代替霉菌孢子,阴性对照组以磷酸缓冲液(PBS)代替霉菌孢子。进行肺组织病理检查,支气管肺泡灌洗液(BALF)中细胞计数分类和总蛋白浓度测定,ELISA测定BALF中的白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)及血浆中的总IgE和特异性IgE,测定小鼠气道阻力。结果模型组和阳性对照组可见明显炎症细胞浸润,特别是嗜酸性粒细胞,而阴性对照组则未见。BALF中细胞总数和嗜酸性粒细胞比例较阴性对照组明显升高(P<0.05);BALF上清中IL-4、总蛋白水平较阴性对照组明显升高(P<0.05),IFN-γ水平较阴性对照组降低(P<0.05)。血浆中总IgE和特异性IgE较阴性对照组明显升高(P<0.05)。小鼠气道阻力较阴性对照相比有明显升高(P<0.05)。结论使用互隔交链孢霉孢子致敏和激发C57BL/6小鼠成功的建立了小鼠气道变态反应性炎症模型。     

磷酸酰肌醇-3激酶抑制剂对小鼠气道炎症影响的实验研究

目的观察磷酸酰肌醇-3激酶(PI3K)抑制剂LY294002对卵白蛋白(OVA)诱导哮喘小鼠气道炎症的影响。方法Balb/c小鼠30只,随机分为三组,每组10只:PI3K抑制剂治疗组(LY组),OVA组和正常对照组(NC组)。通过OVA多次腹腔注射致敏和反复雾化激发,建立哮喘小鼠模型。末次抗原激发后48h收集支气管肺泡灌洗液(BALF)和骨髓标本,计数细胞总数和嗜酸性粒细胞(Eos),并行肺组织学检查。结果与OVA组比较,LY组BALF中细胞总数及Eos绝对数明显减少(p<0.05);肺组织中细支气管、血管周围Eos浸润得到控制,气道粘液分泌减少。结论PI3K抑制剂(LY294002)对卵白蛋白(OVA)致敏的哮喘小鼠气道炎症具有抑制作用。     

复方甘草酸苷对哮喘大鼠气道炎症的影响

目的探讨复方甘草酸苷对哮喘大鼠气道炎症的影响及其机制。方法40只SPF级Wistar大鼠随机分成健康对照组（Ⅰ组）、哮喘模型组（Ⅱ组）、复方甘草酸苷哮喘治疗组（Ⅲ组）、哮喘治疗阴性对照组（Ⅳ组）,10只／组,Ⅱ、Ⅲ、Ⅳ组大鼠用卵清蛋白（OVA）氢氧化铝混合液腹腔注射致敏和OVA超声雾化吸入激发建立哮喘大鼠模型,Ⅲ组大鼠每次激发前1h腹腔注射复方甘草酸苷200μg,Ⅳ组大鼠每次激发前用等体积生理盐水代替复方甘草酸苷腹腔注射,Ⅰ组大鼠腹腔注射用等体积生理盐水代替OVA氢氧化铝混合液,超声雾化吸入用等体积生理盐水代替OVA。所有大鼠在末次激发后6h取中心静脉血2ml测定IgE含量,取左肺支气管肺泡灌洗液（BALF）行细胞计数与分类,并测定BALF中细胞因子IL4、IL-5、IL-13及IFN-γ含量。结果Ⅱ组静脉血IgE含量及BALF中中性粒细胞、淋巴细胞、嗜酸性粒细胞计数和IL-4、IL-5、IL-13含量显著高于Ⅰ组和Ⅲ组（P〈0．01）,IFN-γ含量显著低于Ⅰ组和Ⅲ组（P〈0．01）;Ⅱ组与Ⅳ组各项指标差异均无统计学意义（P〉0．05）;Ⅲ组与Ⅰ组相比,IgE含量及中性粒细胞、淋巴细胞、嗜酸性粒细胞计数和IL4、IL-5及IL-13含量升高,单核巨噬细胞计数和IFN-γ含量降低,但均无统计学意义（P〉0．05）。结论复方甘草酸苷可能通过调节Th1／Th2细胞因子平衡,来抑制Th2细胞免疫学效应,促进Th1细胞免疫学效应,从而减轻气道炎症,降低气道高反应性。     

封闭胸腺基质淋巴生成素受体对超敏原引起的气道炎症性应答的作用

目的为阐明胸腺基质淋巴生成素受体（Thymic stromal lymphopoietin receptor,TSLPR）在超敏原引起的气道炎症应答中的作用,并在小鼠模型中探讨局部封闭TSLPR用以缓解哮喘的可能性。方法对TSLPR抗体或同型抗体预处理,且以鸡卵白蛋白（OVA）诱导的各组小鼠,分别行气道浸润细胞的分类和记数、H＆E和PAS的肺组织染色分析;并以酶联免疫吸附测定法（ELISA）检测支气管肺泡灌洗液中炎性细胞因子;进一步分析OVA激发的小鼠树突状细胞（DCs）的迁移能力和成熟状态。结果在小鼠OVA致敏前施以抗TSLPR抗体可显著减少气道粘液的分泌、嗜酸性粒细胞和淋巴细胞浸润;同时引发IL-4、IL-5水平的明显下降。而作为其机制之一,TSLPR的中和作用可阻止超敏原诱导的DCs的成熟和迁移。结论封闭TSLPR介导的信号通路可缓解哮喘小鼠的气道炎症反应,有望成为一项新的防治气道变应性疾病策略。     

Eosinophilic inflammation in allergic rhinitis and nasal polyposis

On histopathological examination, nasal polyps and nasal mucosa in allergic rhinitis show different forms of pseudostratified respiratory epithelium, whereas the dominant characteristic of lamina propria is an eosinophilic infiltration. The aim of this study was to compare interleukin (IL)-5 and eosinophilic cationic protein (ECP) levels in the nasal fluid of 42 patients: 12 with allergic rhinitis and nasal septal deviation, 17 non-atopic patients with nasal polyposis, and 13 atopic nasal polyp patients were enrolled in this cross-sectional study. Nasal secretion samples were collected a few days before surgery. The levels of IL-5 were measured using flow cytometry and the ECP using a commercial ELISA kit. In addition, we counted eosinophils in hematoxylin-and-eosin-stained sections of all nasal polyp and all nasal mucosa samples taken from the inferior nasal turbinates during septoplasty. A significantly higher concentration of IL-5 was found in the nasal fluid of atopic patients with nasal polyposis than in non-atopic nasal polyp patients (p=0.025) and patients with allergic rhinitis (p=0.05). ECP was higher in atopic nasal polyp patients than in patients with allergic rhinitis (p<0.0001) and than in non-atopic nasal polyp patients (p<0.0001). Polyp eosinophils were higher in atopic' than in non-atopic patients (p<0.0001) and higher than in the mucosa of patients with allergic rhinitis (p<0.0001). These however had significantly more mucosal eosinophils than was found in the polyps of non-atopic patients' (p=0.025). ECP levels in nasal fluid and eosinophil counts in tissue specimens correlated well in all three groups of patients. Our study has shown that atopic nasal polyp patients have a higher level of eosinophilic inflammation than non-atopic patients with nasal polyps and patients with allergic rhinitis.     

视黄酸对大鼠哮喘模型多种炎性因子的影响

目的探讨吸入视黄酸(RA)对急性发作期和慢性持续期哮喘大鼠多种炎症因子影响和作用机制。方法以鸡卵清蛋白(OVA)激发大鼠7d或30 d以制备急性或慢性哮喘模型。在1-7 d或23-30 d期间分别用10μg/ml RA(RA组)、10μg RA+5μg/ml budesonide(RA+BU组)、石蜡油(PO组)雾化吸入治疗1周。急性模型7 d后取血清与肺脏进行检测,慢性模型30 d及随后休息10 d取上清与肺脏进行检测。用免疫荧光、ELISA法检查肺间质细胞因子与血Th因子表达。结果急性发作期PO组胸腺基质淋巴细胞生成素(TSLP)、核因子-κB(NF-κB)、干细胞因子(SCF)表达增加。RA能下调TSLP,但SCF明显上调。慢性持续期PO组TSLP、NF-κB、SCF、IL-4随时间逐步上调,肺感染病灶多,肺泡隔中度增宽。RA组主要炎症因子水平逐渐降低,其中TSLP、IL-4、SCF减少具统计学差异;肺感染病灶少,肺泡隔增宽较轻。BU+RA组下调TSLP表达,但SCF持续上调,肺泡出血与感染较重。结论 RA通过活化巨噬细胞,短暂加重急性发作期哮喘过敏反应,但抑制慢性持续期哮喘肺间质过敏炎症,具免疫调整和抗感染作用。     

Combined vaccination against IL-5 and eotaxin blocks eosinophilia in mice

Interleukin-5 (IL-5) is a cytokine which is essential for the maturation of eosinophils in bone marrow and for their release into the blood. Eotaxin is a CC type chemokine implicated in the recruitment of eosinophils in a variety of inflammatory disorders. Since eosinophil-activity is governed by these two pathways, we targeted both IL-5 and eotaxin by active vaccination to block eosinophilia. We produced two vaccines by chemically cross-linking IL-5 or eotaxin to a virus-like particle (VLP) derived from the bacteriophage Qβ, yielding highly repetitive arrays of these cytokines on the VLP surface. Both vaccines overcame self-tolerance and induced high antibody titers against the corresponding self-molecules in mice. Immunization with either of the two vaccines reduced eosinophilic inflammation of the lung in an ovalbumin (OVA) based mouse model of allergic airway inflammation. Animals immunized with the two vaccines at the same time developed high antibody titers against both cytokines and also reduced eosinophil-infiltration of the lung. These data demonstrate that targeting either IL-5 or eotaxin may lower eosinophilia. Simultaneous immunization against IL-5 and eotaxin demonstrates that such a therapeutic approach may be used to treat complex disorders in which multiple mediators are involved.     

Effect of Cone of Pinus densiflora on DNCB-Induced Allergic Contact Dermatitis-Like Skin Lesion in Balb/c Mice

Cone of Pinus densiflora (CP), or Korean red pinecone, is a cluster of Pinus densiflora fruit. CP has also been verified in several studies to have anti-oxidation, anti-fungal, anti-bacterial, and anti-melanogenic effects. However, anti-inflammatory effects have not yet been confirmed in the inflammatory responses of pinecones to allergic contact dermatitis. The purpose of this study is to prove the anti-inflammatory effect of CP on allergic contact dermatitis (ACD) in vitro and in vivo. CP inhibited the expression of TSLP, TARC, MCP-1, TNF-α, and IL-6 in TNF-α/IFN-γ-stimulated HaCaT cells and MCP-1, GM-CSF, TNF-α, IL-6, and IL-8 in PMACI (phorbol-12-myristate-13-acetate plus A23187)-stimulated HMC-1 cells. CP inhibited the phosphorylation of mitogen-activated protein kinase (MAPKs), as well as the translocation of NF-κB on TNF-α/IFN-γ stimulated in HaCaT cells. In vivo, CP decreased major symptoms of ACD, levels of IL-6 in skin lesion, thickening of the epidermis and dermis, infiltration of eosinophils and mast cells, and the infiltration of CD4⁺ T cells and CD8⁺ T cells. This result suggests that CP represents a potential alternative medicine to ACD for diseases such as chronic skin inflammation.     

CD4＋T细胞及淋巴细胞亚群在小儿哮喘与肺炎中的水平及临床意义

目的：探究CD4＋T细胞及淋巴亚群在小儿哮喘与肺炎中的水平及临床意义。方法选取东营市广饶县中医院儿科自2013年9月至2015年9月收治的34例哮喘患儿作为哮喘组,33例肺炎患儿作为肺炎组,另选择同时期的正常体检小儿30例作为对照组,3组均在早晨空腹情况下抽取静脉血5mL,并于3小时内进行标本处理,后采用流式细胞技术检测外周血中CD4＋T细胞内细胞因子T细胞辅助细胞（Th）1、Th2、Th17及外周血中CD4＋CD25＋FOXP3＋调节性T细胞的水平。结果与对照组相比,哮喘组CD4＋CD25＋FOXP3＋、CD4＋IL-4＋Th1及CD4＋IL-17＋Th17细胞百分比均增加明显,差异均具有统计学意义（ t值分别为5．34、5．89、6．11,均P＜0．05）。与对照组相比,肺炎组CD4＋CD25＋FOXP3＋细胞百分比降低,差异具有统计学意义（ t＝4．56, P＜0．05）。与肺炎组相比,哮喘组CD4＋CD25＋FOXP3＋、CD4＋IL-4＋Th1、CD4＋IL-17＋Th17细胞百分比均增加,差异均具有统计学意义（t值分别为6．12、6．45、6．39,均P＜0．05）。与对照组相比,哮喘组淋巴细胞水平降低,单核细胞、嗜酸性粒细胞水平升高,差异均具有统计学意义（t值分别为5．34、5．26、5．19,均P＜0．05）。与对照组相比,肺炎组单核细胞水平升高,差异具有统计学意义（t＝4．99,P＜0．05）。与肺炎组相比,哮喘组嗜酸性粒细胞水平升高,差异具有统计学意义（t＝5．29,P＜0．05）。结论 CD4＋T细胞及淋巴细胞亚群检测在诊断与鉴别小儿哮喘与肺炎疾病时具有重要临床意义,为临床治疗提供参与依据。