本文選題：肥大細(xì)胞　切入點(diǎn)：細(xì)胞模型　出處：《暨南大學(xué)》2009年碩士論文

【摘要】： [目的]用本實(shí)驗(yàn)室建立的組胺體外釋放肥大細(xì)胞模型,將過(guò)敏原的不同組分置酶標(biāo)板孔中誘發(fā)致敏肥大細(xì)胞釋放組胺,通過(guò)組胺釋放的量,分析過(guò)敏原中不同過(guò)敏原組分的致敏差異性。該方法可作為體外分析過(guò)敏原不同組分的致敏性強(qiáng)弱、確定過(guò)敏原診斷以及抗體芯片的必需組分和評(píng)價(jià)過(guò)敏原疫苗效果。 [方法]蝦粗制過(guò)敏原與塵螨粗制過(guò)敏原分別免疫C57/BL6小鼠,收集小鼠腹腔致敏肥大細(xì)胞(PMC),分別用蝦、塵螨中不同過(guò)敏組分誘導(dǎo)PMC釋放組胺,分別用組胺試劑盒、熒光法檢測(cè)其釋放水平,并進(jìn)行比較。 [結(jié)果]從蝦蛋白中純化出80KD、36KD和21KD等三種蝦致敏蛋白,分別用這三種致敏蛋白誘導(dǎo)PMC釋放組胺,80KD、36KD誘導(dǎo)釋放的組胺最高,21KD蛋白次之,表明80KD、36KD蛋白是蝦致敏蛋白中的主要過(guò)敏原;蝦致敏蛋白混合誘導(dǎo)明顯比單一組分誘導(dǎo)釋放組胺多,并低于致敏蛋白單一誘導(dǎo)的組胺量的疊加,表明蝦蛋白組分在誘導(dǎo)釋放組胺中存在協(xié)同效應(yīng)。塵螨致敏蛋白中,Der f2致敏性強(qiáng)于Der f1,說(shuō)明Der f2是塵螨致敏蛋白中的主要致敏原,塵螨混合誘導(dǎo)PMC釋放組胺時(shí),隨著Der f2比例的升高組胺釋放量也升高。熒光法測(cè)體外釋放組胺量,靈敏、穩(wěn)定、客觀有效。 [結(jié)論]體外肥大細(xì)胞組胺釋放模型能夠客觀反映蝦過(guò)敏原和塵螨過(guò)敏原的不同過(guò)敏原組分釋放組胺的差異性;檢測(cè)組胺釋放量比檢測(cè)血清中IgE或其他物質(zhì)更能反映不同過(guò)敏原組分的致敏強(qiáng)弱;體外檢測(cè)過(guò)敏原模型比體內(nèi)檢測(cè)過(guò)敏原模型更具有可行性、可操作性和可推廣性。該模型可用于過(guò)敏原的體外診斷、疑難過(guò)敏癥的病因分析和過(guò)敏原檢測(cè)或治療性疫苗的評(píng)價(jià),特別是為食品過(guò)敏原的標(biāo)準(zhǔn)化研究等提供新技術(shù)。
[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.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類(lèi)號(hào)】：R392.1

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 宋偉;三種不同來(lái)源的乳蛋白致敏性強(qiáng)弱研究[D];東北農(nóng)業(yè)大學(xué);2011年

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本文編號(hào)：1721276