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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/9307完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃義侑 | |
| dc.contributor.author | Li-Ping Tseng | en |
| dc.contributor.author | 曾莉萍 | zh_TW |
| dc.date.accessioned | 2021-05-20T20:16:53Z | - |
| dc.date.available | 2021-05-20T20:16:53Z | - |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-07-03 | |
| dc.identifier.citation | 1. Alexander DJ. Newcastle disease and other Paramyxoviridae infections. Iowa State University Press 1997:541-569.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/9307 | - |
| dc.description.abstract | 利用微脂粒包覆活的(living)或去活的(killing)新城病毒(NDV)，以點鼻(intranasal)方式接種雞隻，發現在雞隻鼻腔/氣管沖洗液與血清中都有產生特異性NDV抗體。給予雞隻接種三種不同成份的巨型多層微胞微脂粒，其中使用磷脂醯膽鹼(PC)或磷脂醯絲胺酸(PS)組成的微脂粒，在動物實驗中黏膜分泌型抗體(s-IgA)與血清抗體(IgG)及功能性抗體力價(HI titer)表現量在兩次接種後抗體均提升，攻毒後保護率也達80-90%。在微脂粒加入脂多醣體(LPS)佐劑，接種於雞隻後，在利用PC微脂粒組會增加黏膜抗體與PS微脂粒組增加會血清中抗體力價。更進一步，添加銀耳多醣(tremella)以增加微脂粒的黏度，延長微脂粒在黏膜時間，點鼻接種後黏膜分泌型抗體與血清中抗體均顯著提升，攻毒後有100%的保護率。微脂粒佐劑對於免疫細胞之機轉由共軛焦顯微鏡觀察到，PC組成份微脂粒被雞脾臟巨噬細胞胞吞量較多。PC微脂粒與PS微脂粒均活化雞脾臟巨噬細胞產生NO釋放，其中PC微脂粒NO的釋放可被Bay 11-7085抑制與U0126抑制，表示PC微脂粒在早期活化雞脾臟巨噬細胞是以NF-κB磷酸化與ERK上游的MEK磷酸化的路徑。最後我們利用獲得免疫接種微脂粒或銀耳多醣微脂粒的雞隻，發現下游的p-ERK磷酸化的現象存在，其中微脂粒p-ERK磷酸化量較多而銀耳多醣微脂粒p-ERK磷酸化量較少，p-ERK的磷酸化主要促進免疫反應走向體液免疫的Th2路徑，因此，銀耳多醣微脂粒組造成的免疫反應是完整的細胞免疫的Th1與體液免疫的Th2路徑的生成。 | zh_TW |
| dc.description.abstract | Liposomal-NDV vaccine intranasal administration to chicken can induce specific-NDV antibody at mucosal and serum. The adjuvant effect of multi-lamellar vehicles (MLVs) liposomes formulated with three phospholipids including phosphatidylcholine-liposomes (PC-Lip), phosphatidylserine-liposomes (PS-Lip), and stearylamine-liposomes (SA-Lip) was compared with that for virus alone using the inactivated Newcastle disease virus (NDV) as model antigen. PC-Lip and PS-Lip induced more NDV antibody titer after secondary immunizations. In response to virulent viral challenge, all control animals died, whereas 80~90% of animals which still survived. Added LPS with liposomal-NDV vaccine which enhanced chickens produced antibody in serum and nasal/tracheal lavages. Furthermore, we combined tremella with liposomal-NDV vaccine to enhance the muco-adhesive property and prolong liposomes on the mucosal surface. Chickens received T-PC-Lip resulted in a significant increase in HI titer also elicited a significant mucosal secretary immunoglobulin A (s-IgA) response in tracheal lavages and a serum IgG response. After virulent virus challenge, the protection rate of T-PC-Lip vaccine showed 100% survival rate. The molecular mechanism of liposomal-adjuvant was study both in vitro and in vivo. Confocal laser scan microscopy showed that PC-Lip were uptaken into chicken spleen macrophages. PC-Lip and PS-Lip stimulated chickens spleen macrophages produce NO. In response to PC-Lip stimulated, Bay 11-7085 and U0126 inhibited macrophages produced NO. PC-Lip was shown to be involved in both activation of NF-κB and upstream ERK of MEK pathway. Moreover, PC-Lip and T-PC-Lip were shown to be activated ERK pathway in vovo. In conclusion, T-PC-Lip is an active adjuvant for chickens resulting in trigger Th1 pathway of cellular immunity and Th2 pathway of humoral immunity. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-20T20:16:53Z (GMT). No. of bitstreams: 1 ntu-98-D93548009-1.pdf: 2315678 bytes, checksum: a086640eda350a6fef23484309dadb91 (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 口試委員會審定書 ...................................................................................................... i
誌謝 ............................................................................................................................. ii 中文摘要 .................................................................................................................... iii 英文摘要 .................................................................................................................... iv 表目錄 .....................................................................................................................…vi 圖目錄 ....................................................................................................................... vii 第一章 緒言及文獻回顧 ............................................................................................1 1.1 黏膜免疫之介紹 ...................................................................................................1 1.2 動物用黏膜接種之疫苗.........................................................................................2 1.3 佐劑之介紹 ...........................................................................................................3 1.4 顆粒性疫苗傳輸系統 ...........................................................................................6 1.5 微脂粒佐劑 ...........................................................................................................6 1.6 脂多醣體與黏著性多醣體 .................................................................................10 1.7 雞隻黏膜感染病毒 .............................................................................................11 第二章 論文研究目的 ..............................................................................................13 第三章 材料與方法 ..................................................................................................14 3.1 實驗動物 .............................................................................................................14 3.2病毒 .......................................................................................................................14 3.3實驗試藥及分析試劑 ...........................................................................................14 3.3.1試劑藥品..............................................................................................................14 3.3.2抗體 ....................................................................................................................15 3.3.3塑膠、玻璃製品 ................................................................................................16 3. 4微脂粒包覆病毒抗原 ..........................................................................................16 3.4.1 微脂粒配方........................................................................................................16 3.4.2 製備流程 ..........................................................................................................17 3.4.3 粒徑、界面電位與包覆率 ..............................................................................17 3.4.4 相轉移溫度........................................................................................................18 3.4.5 螢光微脂粒 ......................................................................................................18 3.4.6 雙佐劑微脂粒 ..................................................................................................19 3.4.7 銀耳多醣微脂粒和三仙膠微脂粒 ..................................................................19 3.5免疫接種、採血和鼻與氣管沖洗液 ...................................................................20 3.6血球凝集抑制試驗與酵素免疫連結分析法 .......................................................21 3.7攻毒實驗 ...............................................................................................................22 3.8雞脾臟細胞分離 ...................................................................................................22 3.9共軛焦顯微鏡 .......................................................................................................22 3.10 RT-PCR偵測病毒 ..............................................................................................23 3.11材料對細胞之毒性偵測 ....................................................................................24 3.12偵測細胞培養液中NO .......................................................................................24 3.13西方點墨法偵測蛋白質表現 ............................................................................25 第四章 結果 .............................................................................................................27 第五章 討論 .............................................................................................................40 第六章 結論 .............................................................................................................48 參考文獻 ...................................................................................................................49 表 ...............................................................................................................................64 圖 ...............................................................................................................................72 發表論文列表 ...........................................................................................................93 | |
| dc.language.iso | zh-TW | |
| dc.title | 微脂粒佐劑應用於雞點鼻免疫之研究 | zh_TW |
| dc.title | The intranasal delivery study of liposomal-vaccine to SPF chickens | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 李龍湖,劉得任,鍾次文,鄧明中 | |
| dc.subject.keyword | 微脂粒,佐劑,黏膜,新城病毒,一氧化氮,黏膜黏著,銀耳多醣, | zh_TW |
| dc.subject.keyword | liposome,adjuvant,mucosal,NDV,nitric oxide,mucoadhesive,Tremella, | en |
| dc.relation.page | 94 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2009-07-03 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
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