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研究生:林依瑩
研究生(外文):Yi-Ying Lin
論文名稱:發展傳染性華氏囊病疫苗細胞素佐劑
論文名稱(外文):Cytokine adjuvants for infectious bursal disease vaccine
指導教授:邱繡河
指導教授(外文):Shiow-Her Chiou
學位類別:碩士
校院名稱:國立中興大學
系所名稱:獸醫微生物學研究所
學門:獸醫學門
學類:獸醫學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:85
中文關鍵詞:細胞素佐劑
外文關鍵詞:Cytokine adjuvants
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細胞素有免疫調節的功能,因此極具潛力開發為疫苗佐劑。基於此,我們評估以雞重組介白素2 (recombinant chicken interleukin-2, rChIL-2) 與雞重組介白素18 (recombinant chicken interleukin-18, rChIL-18) 做為傳染性華氏囊病 (infectious bursal disease, IBD) 疫苗佐劑的效力。IBD是由傳染性華氏囊病毒 (infectious bursal disease virus, IBDV) 感染所引起的高度傳染性疾病,此病會造成雛雞免疫抑制,因此對於全世界的畜產業有相大的威脅。為了探討施打rChIL-2、rChIL-18、rChIL-2 與 rChIL-18,對於IBD疫苗免疫後的差異,我們檢測比較各組雞隻在施打疫苗後的免疫反應。依據抗體反應、細胞素表現情形、組織學分析來評估各細胞素佐劑效力。實驗結果顯示,疫苗免疫時同時給予rChIL-2 (1 micro-g)或rChIL-2/rChIL-18 (0.1/1 micro-g)細胞素,可增強雞隻體液性免疫反應,提升IL-2受體alpha鏈 (CD25)、丙型干擾素 (interferon-gamma,IFN-gamma)、誘導型一氧化氮合成酶 (inducible nitric oxide synthase,iNOS) 的基因表現,並減輕雞隻在受IBDV攻擊後華氏囊受損的情形。總結以上研究,使用雞重組介白素2及雞重組介白素18皆可增強傳染性IBD疫苗的免疫原性,可視為好的疫苗佐劑。
Cytokines have been considered as potential candidates for vaccine adjuvants based on their immunoregulatory roles. In this study, we evaluated the efficacy of recombinant chicken interleukin-2 (rChIL-2) and recombinant chicken interleukin-18 (rChIL-18) as vaccine adjuvants to combat against the infectious bursal disease virus (IBDV). Infectious bursal disease (IBD) is an acute contagious viral disease caused by infections of IBDV, resulting in severe immunosupression in young chickens, and therefore threatens to the poultry worldwide. After immunization of IBD vaccine with or without coadministration of rChIL-2, rChIL-18, and rChIL-2 plus rChIL-18, we examined and compared immune responses among vaccinated chickens. The adjuvancity of cytokines was assessed based on antibody responses, cytokine expressions, and histological analyses. Vaccination with rChIL-2 (1 micro-g) or rChIL-2/rChIL-18 (0.1/1 micro-g) significantly enhanced humoral immune response, upregulated gene expression of CD25, IFN-gamma, and inducible nitric oxide synthase (iNOS), and reduced lesions of bursa after IBDV challenge. In conclusion, both rChIL-2 and rChIL-18 may serve as good adjuvants to enhance the immunogenicity of IBD vaccine.
目次------------------------------------------------------------------------ I
中文摘要-------------------------------------------------------------------III
英文摘要-------------------------------------------------------------------IV
圖表索引-------------------------------------------------------------------V
中英文對照表-------------------------------------------------------------VII
第一章 序言-------------------------------------------------------------1
第二章 文獻探討-------------------------------------------------------3
第一節 傳染性華氏囊病 (infectious bursal disease, IBD)-3
1-1 IBD簡介------------------------------------------------- 3
1-2 IBD病史--------------------------------------------------3
1-3 IBDV特性------------------------------------------------4
1-4 IBDV抗原性與致病性---------------------------------5
1-5 IBD病徵與防疫-----------------------------------------5
第二節 細胞素 (cytokine)----------------------------------------7
2-1 細胞素簡介-----------------------------------------------7
2-2 介白素2 (interleukin-2, IL-2)--------------------------7
2-2-1 IL-2的發現-----------------------------------------7
2-2-2 IL-2生物特性--------------------------------------8
2-2-3 IL-2目前的應用-----------------------------------10
2-3 雞介白素2 (chicken IL-2, ChIL-2) 簡介------------10
2-3-1 ChIL-2的發現-------------------------------------10
2-3-2 ChIL-2生物特性----------------------------------11
2-4 介白素18 (interleukin-18, IL-18) 簡介--------------11
2-4 IL-18的發現----------------------------------------------12
2-4-2 IL-18生物特性-------------------------------------12
2-4-3 IL-18的應用----------------------------------------14
2-5 雞介白素18 (chicken IL-18, ChIL-18)----------------15
2-5-2 ChIL-18的發現與生物特性----------------------15
第三節 疫苗佐劑開發----------------------------------------------15
第三章 材料與方法-------------------------------------------------------19
第四章 結果-----------------------------------------------------------------30
4-1 重組蛋白純化----------------------------------------------------30
4-1-1 rChIL-2純化----------------------------------------------30
4-1-3 rChIL-18純化---------------------------------------------30
4-2 細胞素安全劑量測試與有效劑量評估-----------------------31
4-3 rChIL-2、rChIL-18做為傳染性華氏囊病疫苗佐劑效力評估 I 32
4-3-1 偵測CD25、IFN-gamma基因表現與血清中抗體力價--------- 32
4-3-2 華氏囊組織病理評估-------------------------------------34
4-3-3 臨床徵狀----------------------------------------------------35
4-3-4 綜觀細胞素疫苗佐劑效力------------------------------- 35
4-4 rChIL-2、rChIL-18 做為傳染性華氏囊病疫苗佐劑效力評估II 37
4-4-1 Th1 相關基因表現----------------------------------------- 37
4-4-2 檢測血清中IBDV抗體力價----------------------------- 38
4-4-3 檢測血清中ND抗體力價-------------------------------- 40
4-4-4 華氏囊組織病理分析-------------------------------------- 41
第五章 討論-------------------------------------------------------------------- 44
圖表-------------------------------------------------------------------------------- 52
參考文獻-------------------------------------------------------------------------- 78
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