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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 龐飛(Victor Fei Pang) | |
| dc.contributor.author | Chuan-Chuan Lin | en |
| dc.contributor.author | 林嫥嫥 | zh_TW |
| dc.date.accessioned | 2021-05-16T16:24:43Z | - |
| dc.date.available | 2015-07-19 | |
| dc.date.available | 2021-05-16T16:24:43Z | - |
| dc.date.copyright | 2013-07-19 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-06-19 | |
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Yu, S., Opriessnig, T., Kitikoon, P., Nilubol, D., Halbur, P.G., Thacker, E., 2007. Porcine circovirus type 2 (PCV2) distribution and replication in tissues and immune cells in early infected pigs. Vet. Immunol. Immunopathol. 115, 261-272. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/6273 | - |
| dc.description.abstract | 黃麴毒素B1(AFB1)是常見於亞熱帶及熱帶地區飼料原料穀物的黴菌毒素，豬隻可能因攝食到黴菌汙染的飼料而接觸AFB1，由於長期接觸低劑量AFB1會影響動物的免疫系統，因此AFB1可能成為豬隻產生豬環狀病毒相關症(PCVAD)的環境因子。本研究自無臨床症狀的豬第二型環狀病毒(PCV2)感染豬隻分離周邊血液單核細胞(PBMCs)，並將細胞進一步分化為單核球來源的樹突細胞(MoDCs)以做為抗原呈獻細胞(APCs)的研究模式，檢測豬免疫細胞於in vitro條件下與AFB1共同後培養，對其細胞功能及PCV2複製的影響。在對免疫細胞功能影響的結果顯示，10 μg/ml 高濃度AFB1處理72小時會減少PBMCs受到致裂原Con A刺激後之細胞存活率及細胞增殖能力；10 μg/ml高濃度AFB1處理48小時會使MoDCs變小、變圓，並失去樹突狀結構，並會降低MoDCs的抗原攝取與處理能力；10 μg/ml高濃度AFB1處理24小時後會增加MoDCs的細胞表面抗原CD40、CD83及CD86，以及免疫抑制性細胞激素IL-10的 mRNA表現量；0.001 μg/ml 低濃度AFB1則在處理後6小時造成MoDCs的CD40及CD83 mRNA表現量上升。在對PCV2複製的影響上，PBMCs受到72小時致裂原Con A的活化下，雖然10 μg/ml AFB1處理並不影響整個處理單位內細胞中的總病毒量，但卻可能增加單位細胞內所含的PCV2病毒量；而MoDCs經過10 μg/ml AFB1處理72小時後，也有處理單位內細胞量減少但細胞中總病毒量與對照組含量相近的現象。本研究結果顯示10 μg/ml 高濃度AFB1有機會增加MoDCs及Con A活化之PBMCs單位細胞內的PCV2病毒複製、傷害細胞媒介性免疫中的APCs功能及細胞增殖能力，並與PCV2同樣能刺激細胞的IL-10 mRNA表現而造成免疫抑制，故可能促使攝食到飼料被高量AFB1汙染的PCV2次臨床感染豬隻發展為PCVAD患豬。 | zh_TW |
| dc.description.abstract | Aflatoxin B1 (AFB1) is a mycotoxin commonly found in cereal grain and related products in subtropical and tropical regions, and pigs may expose to this toxin through contaminated feed. Chronic, low-dose exposure of AFB1 is known to affect the immunity, and may be a potential environmental factor in association with porcine circovirus type 2 (PCV2) in the induction of porcine circovirus-associated disease (PCVAD). In this study, peripheral blood mononuclear cells (PBMCs) were isolated from healthy subclinically PCV2-infected pigs and further differentiated to monocyte-derived dendritic cells (MoDCs) as an antigen-presenting cells (APCs) model to investigate the changes in immunocyte functions and PCV2 replication after in vitro AFB1 exposure. The PBMCs viability and Con A-induced proliferation were impaired after treated with AFB1 at 10 μg/ml for 72 hours. The cell morphology of MoDCs became smaller or rounded, contained shorter dendrites, and lost the antigen uptaking and processing abilities after exposure to 10 μg/ml AFB1 for 48 hours. The mRNA expression levels of surface markers CD40, CD83, and CD86 and regulatory cytokine IL-10 increased in MoDCs after treated with 10 μg/ml AFB1 for 24 hours. A significantly up-regulated CD40 and CD83 mRNA levels were observed in MoDCs after treated with 0.001μg/ml AFB1 for 6 hours. The cell-based PCV2 load appeared to increase in 10 μg/ml AFB1-treated Con A co-treated PBMCs, although no significant difference was seen in the total copy number of PCV2 genome; similar increase in cell-based PCV2 load might also occurr in 10 μg/ml AFB1-treated MoDCs. These results indicate that a relatively higher concentration of AFB1 such as 10 μg/ml may have the potential to increase PCV2 replication in PBMCs and MoDCs, impair the APCs cell function and cell proliferation in cell-mediated immunity, and cause immunosuppression through enhancement of IL-10 mRNA expression. Thus, it is suggested that AFB1 may potentiate PCVAD development in subclinically PCV2 infected pigs through dietary exposure to AFB1. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-16T16:24:43Z (GMT). No. of bitstreams: 1 ntu-102-R00644002-1.pdf: 5922532 bytes, checksum: e6bd793cca7dfb3c7f29dd8990fde7f4 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 中文摘要 i
英文摘要 ii 目錄 iii 表次 vii 圖次 viii 簡寫對照表 x 第一章 序言 1 第二章 文獻回顧 3 第一節 黃麴毒素 3 1-1 黃麴毒素的生成與常見病徵 3 1-2 黃麴毒素B1的主要毒性及作用機制 4 1-3 氣候變遷與黃麴毒素的關係 5 第二節 黃麴毒素對免疫的影響 6 2-1 黃麴毒素對免疫的影響 6 2-2 活體試驗中黃麴毒素對免疫的影響 7 2-3 黃麴毒素對細胞激素的影響 8 2-4 黃麴毒素對巨噬細胞的影響 8 2-5 黃麴毒素對病毒的影響 9 第三節 豬單核球來源的樹突狀細胞研究模式 10 3-1 豬單核球來源的樹突狀細胞簡介 10 3-2 黴菌毒素對單核球來源的樹突狀細胞(MoDCs)的影 12 響 第四節 豬環狀病毒相關症 13 4-1 豬環狀病毒及離乳後多系統消耗症後群 13 4-2 豬環狀病毒相關症 14 第五節 豬環狀病毒相關症的免疫調控 15 5-1 離乳後多系統消耗症候群(PMWS)的免疫學變化 15 5-2 豬體內豬第二型環狀病毒(PCV2)病毒量與PMWS發 16 病的關係以及致裂原concanavalin A (Con A)刺激模式 5-3 豬第二型環狀病毒與先天性免疫系統(innate immune 17 system)的交互作用 5-4 豬第二型環狀病毒對細胞激素的影響 19 第三章 材料與方法 22 第一節 實驗設計 22 1-1 黃麴毒素對於豬周邊血液單核細胞之功能及豬環狀 22 病毒病毒量之影響 1-2 黃麴毒素對於豬單核球來源的樹突狀細胞之功能及 23 豬環狀病毒病毒量之影響 第二節 實驗材料 24 2-1 實驗動物與採血 24 2-2 黃麴毒素B1、致裂原concanavalin A (Con A)與脂多 24 醣(lipopolysaccharide, LPS)泡製 2-3 細胞培養液 24 2-4 血球分離及清洗用之溶液 25 2-5 核酸萃取及聚合酶鍊鎖反應之藥品 26 2-6 流式細胞儀相關藥品 28 2-7 儀器 28 第三節 實驗方法 28 3-1 細胞的分離及培養模式之建立 28 3-1.1 豬周邊血液單核細胞的分離與純化 28 3-1.2 豬單核球來源的樹突狀細胞的分離與純化 29 3-2 黃麴毒素B1及致裂原Con A對周邊血液單核細胞 (PBMCs)的影響 29 3-2.1 以propridium iodide (PI)染色檢測細胞存活率 30 3-2.2 以CFSE染色檢測細胞增生能力(blastogenesis) 30 3-2.3 以Real-Time PCR偵測豬第二型環狀病毒(PCV2)量 31 3-3 黃麴毒素B1及脂多糖(LPS)對樹突狀細胞的影響 32 3-3.1 倒立顯微鏡觀察形態學變化 32 3-3.2 以細胞表面抗原CD14染色檢測單核球來源細胞純 度 32 3-3.3 以propridium iodide染色檢測細胞存活率 33 3-3.4 以DQ™ ovalbumin檢測MoDCs攝取及處理抗原能 33 力(antigen uptaking and processing) 3-3.5 MoDCs表面抗原及細胞激素mRNA表現量 34 3-3.6 以Real-Time PCR偵測PCV2含量 36 3-4 統計分析 36 第四章 結果 38 第一節 黃麴毒素B1及致裂原Con A對豬周邊血液單核細 38 胞的影響 1-1 豬周邊血液單核細胞(PBMCs)的純化 38 1-2 細胞存活率 38 1-3 細胞增生能力(blastogenesis) 39 1-4 豬第二型環狀病毒量變化情形 40 第二節 黃麴毒素B1及脂多醣對豬單核球來源樹突狀細胞 41 (MoDCs)的影響 2-1 豬MoDCs的純化 41 2-2 MoDCs細胞形態學變化 41 2-3 MoDCs細胞存活率、細胞大小及顆粒性變化 43 2-4 MoDCs抗原攝取及處理能力 43 2-5 MoDCs細胞表面抗原及細胞激素mRNA表現的變化 44 2-6 豬第二型環狀病毒量變化情形 49 第五章 討論 64 第六章 參考文獻 75 附錄 88 | |
| dc.language.iso | zh-TW | |
| dc.title | 黃麴毒素B1對豬免疫細胞及豬第二型環狀病毒細胞內複製的影響 | zh_TW |
| dc.title | The Effect of Aflatoxin B1 (AFB1) on Swine Immunocytes and Intracellular Porcine Circovirus Type 2 (PCV2) Replication | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 鄭謙仁(Chian-Ren Jeng) | |
| dc.contributor.oralexamcommittee | 張志成(Chih-Cheng Chang) | |
| dc.subject.keyword | 黃麴毒素B1,豬周邊血液單核細胞,豬單核球來源的樹突細胞,豬第二型環狀病毒,豬環狀病毒相關症, | zh_TW |
| dc.subject.keyword | Aflatoxin B1,porcine peripheral blood mononuclear cells,porcine monocyte-derived dendritic cells,porcine circovirus type 2,porcine circovirus-associated diseases, | en |
| dc.relation.page | 89 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2013-06-20 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 分子暨比較病理生物學研究所 | zh_TW |
| 顯示於系所單位： | 分子暨比較病理生物學研究所 | |
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