体外释放组胺肥大细胞模型在过敏原分析中的应用

发布时间：2018-04-08 10:48

本文选题：肥大细胞　切入点：细胞模型　出处：《暨南大学》2009年硕士论文

【摘要】： [目的]用本实验室建立的组胺体外释放肥大细胞模型,将过敏原的不同组分置酶标板孔中诱发致敏肥大细胞释放组胺,通过组胺释放的量,分析过敏原中不同过敏原组分的致敏差异性。该方法可作为体外分析过敏原不同组分的致敏性强弱、确定过敏原诊断以及抗体芯片的必需组分和评价过敏原疫苗效果。 [方法]虾粗制过敏原与尘螨粗制过敏原分别免疫C57/BL6小鼠,收集小鼠腹腔致敏肥大细胞(PMC),分别用虾、尘螨中不同过敏组分诱导PMC释放组胺,分别用组胺试剂盒、荧光法检测其释放水平,并进行比较。 [结果]从虾蛋白中纯化出80KD、36KD和21KD等三种虾致敏蛋白,分别用这三种致敏蛋白诱导PMC释放组胺,80KD、36KD诱导释放的组胺最高,21KD蛋白次之,表明80KD、36KD蛋白是虾致敏蛋白中的主要过敏原;虾致敏蛋白混合诱导明显比单一组分诱导释放组胺多,并低于致敏蛋白单一诱导的组胺量的叠加,表明虾蛋白组分在诱导释放组胺中存在协同效应。尘螨致敏蛋白中,Der f2致敏性强于Der f1,说明Der f2是尘螨致敏蛋白中的主要致敏原,尘螨混合诱导PMC释放组胺时,随着Der f2比例的升高组胺释放量也升高。荧光法测体外释放组胺量,灵敏、稳定、客观有效。 [结论]体外肥大细胞组胺释放模型能够客观反映虾过敏原和尘螨过敏原的不同过敏原组分释放组胺的差异性;检测组胺释放量比检测血清中IgE或其他物质更能反映不同过敏原组分的致敏强弱;体外检测过敏原模型比体内检测过敏原模型更具有可行性、可操作性和可推广性。该模型可用于过敏原的体外诊断、疑难过敏症的病因分析和过敏原检测或治疗性疫苗的评价,特别是为食品过敏原的标准化研究等提供新技术。
[Abstract]:[objective] using the mast cell model established in our laboratory to release mast cells in vitro, we divided the different components of allergens into enzyme labeled plate holes to induce the release of histamine and the amount of histamine released by histamine.To analyze the allergy difference of different allergen components in allergen.This method can be used to analyze the sensitivities of different components of allergen in vitro, to determine the essential components of allergen diagnosis and antibody chip and to evaluate the effect of allergen vaccine.[methods] C57/BL6 mice were immunized with crude allergens and coarse allergens of Dermatophagoides dust mites respectively. The mast cells were collected from the abdominal cavity of mice. PMC was induced to release histamine by different allergy components of shrimp and dust mites, and histamine kit was used respectively.The release level was detected by fluorescence method and compared.[results] three kinds of shrimp sensitized proteins were purified from shrimp protein, such as 80KD 36KD and 21KD, respectively. The highest histamine protein of 80KD36KD was released by PMC, the highest was 21KD protein, which indicated that 80KD / 36KD protein was the main allergen in shrimp sensitizing protein.The mixture induction of shrimp sensitized protein was significantly more than that of single component in inducing the release of histamine, and it was lower than the sum of histamine induced by single sensitized protein, which indicated that the protein component of shrimp had synergistic effect in inducing the release of histamine.Fluorescence assay is sensitive, stable and objective to determine the amount of histamine released in vitro.[conclusion] in vitro mast cell histamine release model can objectively reflect the difference of histamine release from different allergen components of shrimp allergen and dust mite allergen.Detection of histamine release is more effective than detecting IgE or other substances in serum to reflect the sensitizing intensity of different allergen components, and in vitro detection of allergen model is more feasible, operable and extensible than in vivo detection of allergen model.The model can be used for the in vitro diagnosis of allergens, the etiological analysis of difficult allergies and the evaluation of allergen detection or therapeutic vaccines, especially for the standardization of food allergens.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392.1

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