本文選題：slgE + IgG��； 參考：《暨南大學(xué)》2006年碩士論文

【摘要】：[目的] 建立食品過(guò)敏原的豚鼠過(guò)敏癥模型，研究海蝦、大豆和腰果等水提粗制蛋白浸液免疫原的免疫原性；其抗血清體外致敏正常豚鼠肥大細(xì)胞，建立過(guò)敏原體外定向釋放組胺，探明過(guò)敏癥發(fā)生過(guò)程中致肥大細(xì)胞脫顆粒的抗體類(lèi)型；Western blot鑒定分析三種食物過(guò)敏原中主要的過(guò)敏原成分。 [方法] 以海蝦、大豆和腰果等水提粗制蛋白浸液作為免疫原，以氫氧化鋁為佐劑免疫豚鼠，建立豚鼠過(guò)敏模型；分離正常豚鼠腹腔肥大細(xì)胞，將其分別與正常和海蝦致敏的抗血清及其分離純化的sIgE、IgG1和IgG孵育，在體外用過(guò)敏原分別激發(fā)，HPLC組胺檢測(cè)法檢測(cè)肥大細(xì)胞的組胺釋放率；應(yīng)用三種食品的水提粗制蛋白浸液的豚鼠抗血清，Western blot分析主要的過(guò)敏原組分和它們的交叉抗原，并用DEAE交換層析分離純化海蝦過(guò)敏原。 [結(jié)果] 海蝦、大豆和腰果蛋白組分等幾種主要食品過(guò)敏原免疫組豚鼠，取致敏豚鼠血清，用ELISA方法檢測(cè)其sIgE水平，發(fā)現(xiàn)致敏后第14d豚鼠血清sIgE效價(jià)最高，分別達(dá)1：80，1：40和1：80。 將正常豚鼠腹腔肥大細(xì)胞與分離純化的海蝦免疫的豚鼠全血清的sIgE、IgG1及IgG分別結(jié)合(致敏)，，在體外用相應(yīng)過(guò)敏原激發(fā)，檢測(cè)組胺定向釋放率，發(fā)現(xiàn)IgE和IgG均參與了豚鼠過(guò)敏激發(fā)，組胺定向釋放率分別為40.05±1.98％和71.43±3.47％；進(jìn)一步研究發(fā)現(xiàn)這種作用是由免疫血清中IgG1亞類(lèi)介導(dǎo)的，而其他IgG亞類(lèi)未參與。 Western blot鑒定海蝦主要過(guò)敏組分的分子量為21、36、38、40kD，其中38kD蛋白與牡蠣浸液呈交叉反應(yīng)；腰果主要過(guò)敏原組分為55kD、28kD和14kD，其中14和28kD與大豆蛋白浸液呈交叉發(fā)應(yīng)：大豆主要過(guò)敏組分為14、28、30、60kD。 [結(jié)論] 用海蝦、大豆和腰果等水提粗制蛋白浸液作為免疫原，建立了食物豚鼠過(guò)敏模型，采用HPLC組胺檢測(cè)法檢測(cè)過(guò)敏原體外定向釋放的組胺，發(fā)現(xiàn)sIgE和IgG1均可參與體外致敏豚鼠肥大細(xì)胞的組胺釋放。Western blot鑒定三種食物中的過(guò)敏原組分，初步確定它們的主要過(guò)敏原組分，并發(fā)現(xiàn)在海蝦與牡蠣，腰果與大豆存在交叉過(guò)敏原。體外致敏肥大細(xì)胞組胺釋放率可作為過(guò)敏原鑒定與評(píng)價(jià)的技術(shù)。
[Abstract]:[objective] to establish an allergic guinea pig model of food allergen, to study the immunogenicity of crude protein extracts from sea shrimp, soybean and cashew nuts, and to sensitize normal guinea pig mast cells by antiserum in vitro. To establish the directional release of histamine from allergens in vitro, and to identify the type of antibodies to degranulate mast cells during anaphylactogenesis. Western blot was used to identify and analyze the main allergens in three kinds of food allergens. [methods] prawns were used. A guinea pig allergic model was established by immunizing guinea pigs with aluminum hydroxide as adjuvant and crude protein extracts from soybean and cashew nuts were used as immunogen, and normal guinea pig peritoneal mast cells were isolated. It was incubated with antiserum sensitized by normal and shrimp and purified IgG1 and IgG respectively. Histamine release rate of mast cells was detected by HPLC histamine assay by allergen stimulation in vitro. The main allergen components and their cross antigens were analyzed by Western blot with the guinea pig antiserum extracted from water extract crude protein extract of three kinds of food, and the allergen of sea shrimp was isolated and purified by DEAE exchange chromatography. [results] the sea shrimp was isolated and purified by DEAE exchange chromatography. The sensitized guinea pigs were immunized with soybean and cashew protein components. The serum of sensitized guinea pigs was detected by Elisa. It was found that the titer of Siga E was the highest in guinea pigs on the 14th day after sensitization. The normal guinea pig peritoneal mast cells were combined with Siga IgG1 and IgG in the whole serum of guinea pigs immunized with purified sea shrimp, respectively. The specific release rate of histamine was detected by using the corresponding allergens in vitro. It was found that both IgE and IgG were involved in the allergic stimulation of guinea pigs, and the specific release rates of histamine were 40.05 鹵1.98% and 71.43 鹵3.47, respectively. The molecular weight of the main allergy components of shrimp identified by Western blot was 21 ~ 366 ~ 38 ~ 40kD, in which 38 KD protein was cross-reacted with oyster extract. The main allergen components of cashew nuts were 55 kD, 28 KD and 14 KD, among which 14 and 28 KD were cross-distributed with soybean protein extract. The main allergy components of soybean were 14 ~ 2830 ~ 60 KD. [conclusion] crude protein extracts from sea shrimp, soybean and cashew nut were used as immunogen. A food guinea pig allergic model was established. Histamine released by allergen in vitro was detected by HPLC. It was found that Siga E and IgG1 were both involved in histamine release from mast cells of guinea pig sensitized in vitro. Western blot was used to identify the allergen components in three foods. Their main allergens were identified and cross allergens were found in shrimp and oysters cashew nuts and soybean. Histamine release rate of mast cells sensitized in vitro can be used as a technique for the identification and evaluation of 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】：2006
【分類(lèi)號(hào)】：R392

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本文編號(hào)：2002625