Identification of a New Allergen from Amaranthus retroflexus Pollen,Ama r 2

BackgroundPollinosis from Amaranthus retroflexus pollen is a common cause of respiratory allergy in Iran with a high positive rate (68.8%) among Iranian allergic patients. The aim of the present study was to evaluate the allergenicity of the A. retroflexus pollen profilin.MethodsUsing sera from twelve patients allergic to A. retroflexus pollen, IgE-binding proteins from the A. retroflexus pollen extract was identified by immunoblotting. The cDNA of A. retroflexus pollen profilin was amplified, then cloned into the pET-21b (+) vector, expressed in Escherichia coli, and finally purified by metal affinity chromatography. The IgE-binding capacity of the recombinant protein was then analyzed by the ELISA, immunoblotting, and inhibition assays, as well as by the skin prick test (SPT).ResultsImmunoblotting results indicated a 14.6 kDa protein with IgE-reactivity to 33% (4/12) among A. retroflexus pollen-allergic patients. Nucleotide sequencing of the cDNA revealed an open reading frame of 399 bp encoding for 133 amino acid residues which was belonged to the profilin family and designated as Ama r 2. A recombinant Ama r 2 (rAma r 2) was then produced in E. coli as a soluble protein which showed a strong IgEreactivity via ELISA confirmed by the SPT. Inhibition experiments revealed high IgE cross-reactivities with the profilins from other plants.ConclusionsThe profilin from the A. retroflexus pollen, Ama r 2, was firstly identified as an allergen. Moreover, rAma r 2 was produced in E. coli as a soluble immunoreactive protein with an IgE-reactivity similar to that of its natural counterpart.     

变应原检测和特异性免疫治疗中不良反应的分析

目的通过对变应原检测和特异性免疫治疗中不良反应的统计分析，了解所用制剂的安全性。方法回顾性分析我院2003年5月至2008年4月变应原检测及2005年5月至2008年4月变应原特异性免疫治疗临床资料，并对不良反应进行统计。采用标准化变应原皮肤点刺液进行变应原检测，使用标准化屋尘螨提取液皮下注射进行特异性免疫治疗。观察变应原检测及特异性免疫治疗中的不良反应发生情况，其严重度分级按欧洲变态反应和临床免疫学会标准。结果在10219例变应原检测中，有26例发生不良反应，占点刺总例数的0．25％，其中17例为2级不良反应，无4级不良反应发生。对166例变应原特异性免疫治疗的患儿，共注射3087人次，有8人，共25人次发生了全身不良反应，占注射总人次的0．81％。其中15例为2级不良反应，无4级不良反应发生。在本组患儿中，发生不良反应的的主要危险因素为高度敏感个体，有4人反复发生不良反应5次，1人反复发生3次。变应原检测显示，尘螨均为强阳性。由于观察细致，处理得当，患儿迅速恢复，无1例致命性不良反应发生。结论采用本文所使用的制剂作变应原检测和特异性免疫治疗，不良反应率均低，且程度不严重，说明其安全性良好。     

变应原吸入激发试验应用概况

支气管哮喘(简称哮喘)是最常见的慢性气道疾病之一.近年来其患病率在全球范围内有逐年增加的趋势.变应性哮喘为哮喘的常见类型,通常于暴露变应原后诱发.环境中的吸入性变应原如尘螨、花粉等为变应性哮喘最主要的致敏物质.通过吸入特定类型及剂量的变应原,对敏感患者进行变应原吸入激发试验,可诱发其产生可控性的哮喘反应,对探讨变应性哮喘的发病机制及新治疗靶点有重要作用.本文就其发展史、原理和操作方法等方面进行阐述.     

布氏菌变应原的研究 2、不同菌种和不同培养方法生产菌素敏感性和化学组成

本文测试了不同菌种的致敏强度,牛种菌>猪种菌>羊种菌,提出皮试阳性标准应有不同。     

植物生物反应器在变应原生产中的研究进展

随着工业化进程的加快,变应性疾病已成为威胁人类健康的严重问题,应用特异性变应原进行脱敏治疗是其惟一有效的办法.新型变应原疫苗的生产研究已成为现今的热门课题之一.近年来随着分子生物学技术的发展,利用转基因植物作为生物反应器生产具有重要经济价值的异源蛋白(特别是医用蛋白)正在成为研究最活跃、产业发展最迅速的热点,该文着重就植物生物反应器近年的发展及其在变应原生产中的应用、优缺点等做一简单综述.     

尘螨Ⅱ类变应原研究进展

尘螨是最常见的室内变应原之一,其最主要的变应原是Ⅰ类变应原和Ⅱ类变应原.该文从尘螨Ⅱ类变应原的研究概况、在螨体内的定位、结构和生化特性研究与同源性和多态性分析、螨性过敏性疾病的致病机制及其疫苗研究等方面进行了综述.     

变应原突变体在支气管哮喘免疫治疗中的应用

特异性免疫治疗（SIT）是对明确了变应原的变态反应性疾病的一种有效的治疗方法。重组变应原及其衍生物的技术的进步大大提高了生产用于治疗变应性疾病的新型疫苗的能力。本文将就用定点突变技术获取的重组变应原突变体在支气管哮喘免疫治疗中的应用做一综述。     

Antisense-mediated silencing of a gene encoding a major ryegrass pollen allergen

Type 1 allergic reactions, such as hay fever and allergic asthma, triggered by grass pollen allergens are a global health problem that affects approximately 20% of the population in cool, temperate climates. Ryegrass is the dominant source of allergens because of its prodigious production of airborne pollen. Lol p 5 is the major allergenic protein of ryegrass pollen, judging from the fact that almost all of the individuals allergic to grass pollen show presence of serum IgE antibodies against this protein. Moreover, nearly two-thirds of the IgE reactivity of ryegrass pollen has been attributed to this protein. Therefore, it can be expected that down-regulation of Lol p 5 production can significantly reduce the allergic potential of ryegrass pollen. Here, we report down-regulation of Lol p 5 with an antisense construct targeted to the Lol p 5 gene in ryegrass. The expression of antisense RNA was regulated by a pollen-specific promoter. Immunoblot analysis of proteins with allergen-specific antibodies did not detect Lol p 5 in the transgenic pollen. The transgenic pollen showed remarkably reduced allergenicity as reflected by low IgE-binding capacity of pollen extract as compared with that of control pollen. The transgenic ryegrass plants in which Lol p 5 gene expression is perturbed showed normal fertile pollen development, indicating that genetic engineering of hypoallergenic grass plants is possible.     

尘螨主要变应原DNA疫苗对其提取液诱导小鼠肺部变应性炎症的免疫保护作用

目的　探讨尘螨主要变应原DNA疫苗对尘螨提取液诱导的小鼠肺部变应性炎症的免疫治疗作用。方法　分别构建表达Derp1和Derp 2的真核表达质粒pCMV Derp 1和pDerp 2 IRES Derp1。 30只BALB/c小鼠随机分为正常组 (6只 )、对照组 (12只 )、单质粒组 (6只 )和混合组 (6只 )。后三组分别用空白质粒、pDerp 2 IRES Derp 1或混合的pDerp 2 IRES Derp 1和pCMV Derp 1免疫 ,4周后用尘螨提取液致敏、激发 ,观察肺部炎症、计分、肺泡灌洗液 (BALF)细胞总数和分类计数 ;酶联免疫吸附法 (ELISA)检测BALF中白细胞介素 4 (IL 4 )和γ干扰素 (IFN γ)水平。结果　细胞总数和嗜酸细胞(EOS)计数 :混合组 [(6± 4 )× 10 5/ml、0 0 5± 0 0 4 ]和对照组 [(2 1± 13)× 10 5/L、1 80± 1 39]比较差异有显著性 (P <0 0 5 ) ;IL 4水平 :混合组 [(16 8± 2 33)g/L]与对照组 [(5 38± 2 5 6 )g/L]比较差异有显著性 (P<0 0 5 ) ,各组间IFN γ比较差异无显著性 (P >0 0 5 )。结论　尘螨主要变应原的DNA疫苗可有效抑制尘螨提取液诱导的肺部变应性炎症 ,能表达两个主要变应原的DNA疫苗其抑制变应性炎症效果优于只表达一种变应原的疫苗。     

空调器隔尘网富集尘螨过敏原的研究

目的 探讨空调隔尘网中尘螨过敏原与哮喘发病的关系。方法 分别从芜湖市区食堂、商场、酒店和居民家中空调器隔尘网上,及其附近空气中采集灰尘样本各10份。用ELISA法检测粉尘螨1类过敏原（Der f 1）、屋尘螨1类过敏原（Der p 1）的浓度及其灰尘提取液过敏原性。结果 芜湖市区食堂、商场、酒店和居民家中空调器隔尘网灰尘中的过敏原Der f 1浓度分别为1.52、1.24、1.31 μg/g 和1.46 μg/g ;Der p 1浓度分别约为1.23、1.12、1.16 μg/g 和1.18 μg/g;2种过敏原在开机1 h后的空气中浓度显著高于开机前（P < 0.05）。ELISA分析结果显示,灰尘提取液中过敏原能与尘螨过敏性哮喘小鼠的IgE抗体产生结合反应。结论 芜湖地区居民空调器隔尘网中含有尘螨1类过敏原,可诱发哮喘。     

黄花蒿花粉新过敏原烯醇化酶鉴定:重组纯化及IgE结合活性

目的 分析并鉴定黄花蒿花粉中新过敏原烯醇化酶以及致敏性.方法 通过对黄花蒿花粉转录组学研究,得到潜在过敏原烯醇化酶的基因序列,通过序列扩增、重组表达和纯化获得重组黄花蒿花粉烯醇化酶,结合过敏患者血清和酶联免疫吸附实验(ELISA)、免疫印迹实验(Western blot)和竞争抑制实验研究黄花蒿花粉烯醇化酶的IgE结合...     

呼出气一氧化氮与肺功能和过敏原检测的相关性分析

目的 探讨呼出气一氧化氮（FeNO）与支气管舒张试验、血中特异性IgE水平的相关性.方法 对我院298例疑似支气管哮喘（简称哮喘）患者的FeNO、支气管舒张试验、血中特异性IgE水平进行统计学分析.结果 298例患者中支气管舒张试验阳性组患者的FeNO值高于阴性组,艾蒿或螨虫组合阳性组的FeNO值高于对应的阴性组.FeNO升高与支气管舒张试验阳性密切相关.结论 FeNO在支气管舒张试验阳性和对艾蒿、螨虫组合阳性的患者中高表达,对哮喘的诊断有一定价值.     

霜天蛾昆虫变应原多克隆抗体的制备及免疫特性分析

目的 制备兔抗霜天蛾变应原多克隆抗体,分析其与特异性过敏患者血清抗体成分的异同,为免疫筛选cDNA文库,制备基因重组霜天蛾变应原奠定基础.方法 应用霜天蛾变应原提取液免疫家兔获得霜天蛾变应原多克隆抗体,用酶联免疫吸附试验(ELISA)法测定混合免疫兔血清抗体效价:应用免疫印迹的方法对比分析免疫兔血清与特异性过敏患者血清抗体成分的异同.结果 混合免疫兔血清效价经ELISA检测,效价>1:10 000,免疫印迹检测兔抗血清免疫球蛋白G相似文献   

过敏性因素在老龄大鼠支气管哮喘模型中的实验研究

目的 探讨过敏性因素在晚发老年哮喘发生中的作用。方法 24只SPF级老年SD大鼠随机分为老年正常对照组（A组）和老年哮喘模型组（B组），每组12只；24只SPF级青年sD大鼠随机分为青年正常对照组（C组）和青年哮喘模型组（D组），每组12只。予卵蛋白腹腔注射并雾化吸入制作SD大鼠支气管哮喘模型；予乙酰胆碱（Ach）行支气管激发试验；以动物呼吸机测定呼气相气道阻力（Re）；测定支气管壁厚度、支气管平滑肌厚度和支气管黏膜和黏膜下炎性细胞数。结果 分别比较2组哮喘大鼠Re，以及2组哮喘大鼠支气管壁厚度、支气管平滑肌厚度和支气管黏膜和黏膜下炎性细胞数，结果显示造模成功；在10—40μg/kg浓度梯度的Ach激发后，B组和D组的Re差异有统计学意义（P〈0．05），在80—160μg／kg浓度梯度的Ach激发后，B组Re增加更明显（P〈0.01），与D组比较，B组支气管壁厚度、支气管平滑肌厚度和支气管黏膜和黏膜下炎性细胞数均明显增加（P〈0．01）。结论 本方法能制备老龄支气管哮喘大鼠模型；变应原对于晚发老年哮喘的发生同样重要。     

哮喘患者肺段过敏原激发试验后淋巴细胞白细胞介素-4受体的变化

目的　了解白细胞介素 4受体 (IL 4R)在过敏性哮喘中的作用。方法　将研究对象分为过敏性哮喘组 (8例 )和对照组 (8例 ) ,哮喘患者行肺段过敏原激发试验 (SAP) ,分离正常人及哮喘患者SAP前和SAP 18h后的外周血单个核细胞 (PBMC) ,取 1× 10 6个细胞培养 2 4h及 48h ,利用流式细胞仪及酶联免疫吸附试验分别测细胞膜IL 4Rα和上清液中的可溶性IL 4Rα。结果　哮喘患者SAP前CD19+ 淋巴细胞IL 4Rα表达阳性率为 (72 .6± 12 .9) % ,明显低于对照组的 (87.9± 5 .6) % ;2 4h培养后CD3 + 及CD4+ 淋巴细胞IL 4Rα表达阳性率分别为 (2 7.2± 8.6) %及 (3 4.3± 11.2 ) % ,明显高于对照组的 (16.4± 6.0 ) %和 (2 0 .0± 8.1) %。SAP后CD3 + 及CD4+ 淋巴细胞IL 4Rα表达阳性率分别为 (2 8.6± 9.8) %及 (3 6.2± 13 .4) %较SAP前的 (17.2± 7.2 ) %和 (2 4.1±9.8) %明显升高 ;且 2 4h培养后PBMC产生可溶性IL 4Rα为 (2 .5 4± 3 .47) pg/ml较SAP前(72 .40± 5 6.86) pg/ml明显减少 (P <0 .0 5 )。 结论　外周血淋巴细胞IL 4Rα的上调及产生可溶性IL 4Rα的能力下降可能在过敏性哮喘发病过程中起重要作用     

Identification of sokotrasterol sulfate as a novel proangiogenic steroid

The potential to promote neovascularization in ischemic tissues using exogenous agents has become an exciting area of therapeutics. In an attempt to identify novel small molecules with angiogenesis promoting activity, we screened a library of natural products and identified a sulfated steroid, sokotrasterol sulfate, that induces angiogenesis in vitro and in vivo. We show that sokotrasterol sulfate promotes endothelial sprouting in vitro, new blood vessel formation on the chick chorioallantoic membrane, and accelerates angiogenesis and reperfusion in a mouse hindlimb ischemia model. We demonstrate that sulfation of the steroid is critical for promoting angiogenesis, as the desulfated steroid exhibited no endothelial sprouting activity. We thus developed a chemically synthesized sokotrasterol sulfate analog, 2beta,3alpha,6alpha-cholestanetrisulfate, that demonstrated equivalent activity in the hindlimb ischemia model and resulted in the generation of stable vessels that persisted following cessation of therapy. The function of sokotrasterol sulfate was dependent on cyclooxygenase-2 activity and vascular endothelial growth factor induction, as inhibition of either cyclooxygenase-2 or vascular endothelial growth factor blocked angiogenesis. Surface expression of alpha(v)beta(3) integrin was also necessary for function, as neutralization of alpha(v)beta(3) integrin, but not beta(1) integrin, binding abrogated endothelial sprouting and antiapoptotic activity in response to sokotrasterol sulfate. Our findings indicate that sokotrasterol sulfate and its analogs can promote angiogenesis in vitro and in vivo and could potentially be used for promoting neovascularization to relieve the sequelae of vasoocclusive diseases.