賈景山臺灣大學:口腔生物科學研究所魏中帆Wei, Chung-FanChung-FanWei2010-05-042018-07-092010-05-042018-07-092009U0001-1307200915083900http://ntur.lib.ntu.edu.tw//handle/246246/178613來自靈芝、酵母菌等真菌細胞壁的多醣類，如以β(1→3), β(1→6)鍵結之化合醣類，統稱為β-聚糖。β-聚糖被報導具有免疫調節的能力，如增進免疫力能預防癌症及微生物感染；或是改變免疫反應的途徑，如降低Th2反應卻增加Th1反應，以減少過敏發生。而鼻腔內給予之疫苗能除了引發接種者血清中特異性免疫球蛋白G (Immunoglobulin G, IgG)，更能使接種者產生在上呼吸道局部的分泌型免疫球蛋白A，IgA在黏膜能中和外來物抗原，降低經飛沬傳染散播的微生物入侵宿主之機率，這是注射疫苗很難達到的效果。並且鼻腔疫苗有容易施行大規模接種之優點，雖然大部分鼻腔疫苗尚在實驗階段，但其具有研發潛力。實驗建立鼻腔內致免的動物模式。以大腸桿菌致病株所分離的忌熱性腸毒素(heat-labile enterotoxin, LT)做為佐劑，卵清白蛋白(ovalbumin, OVA)為抗原，從鼻腔給予BALB/c小鼠。而小鼠從致免前一周開始每周三次分別餵食二次水、熱致死之啤酒酵母或市售啤酒酵母大豆發酵產物直到犧牲。實驗期間，每週採集小鼠的唾液、測定抗白蛋白之分泌型IgA以及採集血清測定抗OVA之IgG和IgE。結果顯示，小鼠餵食啤酒酵母及市售啤酒酵母大豆發酵產物的小鼠唾液中有較高的抗OVA特異性唾液IgA及血清中IgG和IgE。初步認為飲食中含有啤酒酵母及市售啤酒酵母大豆發酵產物可促進特定免疫球蛋白生成，可能透過Th1之細胞激素IFN-γ和Th2之細胞激素IL-5的分泌增加。因此服用熱致死啤酒酵母的相關產品可能加強宿主保護力，增進鼻腔內疫苗效果，但亦有增加對特定抗原過敏之風險。β-glucans as β(1→3), β(1→6) binding polysaccharides derived from fungal cell wall, like Ganoderma lucidum and yeasts. β-glucans have been reported that confers immunomodulation with oral administration or intravenous injection. The immunomodulated functions of β-glucans may promote lymphocytes tumor cytotoxic ability or protect against microbial infections. Nevertheless, some kinds of β-glucan can also skew Th1/Th2 balance, may inhibit allergic response. Intranasal administrated nasal vaccines elicit hosts’ systemic specific immunoglobulin G (IgG), also elicit scretory IgA that may neutralize respiratory pathogens at local mucosa. Nasal vaccines are potential, since they are easily accessible for large population, and required without needles and syringes. In present research, we nasal immunized BALB/c mice with heat-labile enterotoxin(LT) from wild-type entero-pathogenic E. coli and ovalbumin (OVA) for 4 times, 1-week internal. In parallel, we oral administrated the mice with sterile water, heat-killed sacchromyces cerevisiae, or commercial sacchromyces cerevisiae legume fermentated product trice a week, started from 1 week before the first nasal immunization, and last to the second week after the last immunization, mice were sacrificed. Saliva, serum of mice was collected every week since feeding started, the samples were detected the specific anti-OVA IgA, IgG and IgE with ELISA. The present data shows that anti-OVA salivary IgA and anti-OVA serum IgG1, IgG2a and IgE in sacchromyces cerevisiae and commercial sacchromyces cerevisiae legume fermentated product-feeding mice with LT+OVA immunized. It’s preliminarily considered that sacchromyces cerevisiae or related products-intake may enhance specific antibody production, through IL-5 and IFN-γ-secreting increased. Therefore, oral intake sacchromyces cerevisiae or related products may protect hosts against pathogen, enhance nasal vaccine efficacy, but increase IgE-mediated allergy risk.中文摘要.........................................................................................................................i文摘要.........................................................................................................................ii目錄.............................................................................................................................iii一章 文獻回顧...........................................................................................................1一節 黏膜免疫.....................................................................................................1二節 鼻腔免疫與鼻腔相關淋巴組織免疫.........................................................3三節 腸道免疫.....................................................................................................4四節 健康食品的免疫調節.................................................................................5五節 啤酒酵母.....................................................................................................6六節 健康食品之免疫調節評估方法.................................................................8七節 研究動機與目的.........................................................................................8二章 材料及方法.....................................................................................................10一節 實驗動物小鼠...........................................................................................10二節 製備熱致死啤酒酵母...............................................................................10三節 餵食與黏膜致免計畫...............................................................................10四節 檢體與組織收集.......................................................................................11五節 酵素連結免疫吸附試驗...........................................................................12六節 細胞增生測試...........................................................................................14七節 細胞分群測定...........................................................................................14八節 聚丙烯醯胺膠體電泳法及銀染...............................................................14九節 統計方法...................................................................................................15三章 實驗結果.........................................................................................................16一節 鼻腔致免溶液進行膠片電泳...................................................................16二節 小鼠體重比較...........................................................................................16三節 唾液、BALF、血清中抗OVA之特異性IgA.............................................16四節 血清中抗OVA之特異性IgG與IgE.........................................................17五節 唾液、BALF、血清中總IgA含量..............................................................18六節 血清中總IgG和IgE..................................................................................19七節 脾臟及不同淋巴結細胞的特異性增生...................................................20八節 脾臟及不同淋巴結細胞的非特異性增生...............................................21九節 初代細胞針對OVA分泌的細胞激素......................................................22十節 初代免疫細胞分群之百分比...................................................................23四章 討論.................................................................................................................23一節 唾液、BALF、血清中抗OVA之特異性IgA與細胞激素......................23二節 血清中抗OVA之特異性IgG與IgE與細胞激素....................................24三節 血清中總抗體含量...................................................................................25四節 胰臟及淋巴結細胞的增生與細胞表現型之百分比...............................26五節 小鼠體重比較...........................................................................................26五章 總結論.............................................................................................................27六章 參考文獻.........................................................................................................29application/pdf718706 bytesapplication/pdfen-US免疫調節鼻腔內致免唾液免疫球蛋白A血清免疫球蛋白immunomudulationintranasal immunizationsalivary IgAserum immunoglobulins[SDGs]SDG3http://ntur.lib.ntu.edu.tw/bitstream/246246/178613/1/ntu-98-R96450012-1.pdf