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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 龐飛(Victor Fei Pang) | |
dc.contributor.author | Chun-Ming Lin | en |
dc.contributor.author | 林俊明 | zh_TW |
dc.date.accessioned | 2021-06-13T05:47:53Z | - |
dc.date.available | 2009-07-27 | |
dc.date.copyright | 2006-07-27 | |
dc.date.issued | 2006 | |
dc.date.submitted | 2006-07-09 | |
dc.identifier.citation | Adair, B.M., 2000, Immunopathogenesis of chicken anemia virus infection. Dev Comp Immunol 24, 247-255.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/33871 | - |
dc.description.abstract | 豬隻離乳後多系統消耗性症候群(PMWS)是豬隻新興的疾病,其主要病原為豬第二型環狀病毒(PCV2),並且以免疫系統內的淋巴球流失以及組織球浸潤為其主要病理特徵。截至目前為止,PCV2是否會於免疫系統內複製以及PMWS的致病機制仍不甚清楚。在本實驗中,我們建立了以致裂原刺激豬隻周邊血液淋巴球(PBL)的PCV2感染模式。首先,在第一階段實驗中針對健康無PCV2帶原的豬隻進行測試,當同時對其PBL以致裂原刺激與PCV2攻毒處理後,其PBL中僅0.1%的細胞呈現PCV2抗原陽性;藉由MTT與blastogenesis分析,均顯示PCV2可以降低PBL對於致裂原刺激的反應,然而PCV2對於PBL的存活率或細胞凋亡率則無顯著影響。在第二階段的實驗中,針對臨床上健康的PCV2帶原豬隻進行測試,結果顯示經由免疫螢光染色或原位雜交的方式,PCV2的抗原或核酸可以顯著地在致裂原刺激後的PBL中觀測得到;在致裂原刺激後第4天,PCV2抗原陽性率於PBL中最高可達20%,其中以表面抗原呈現IgM+的細胞最具感受性,病毒力價亦有顯著上升的情況;此外,PCV2 ORF1蛋白的表現通常位於細胞核的周邊或同時出現於核内,而ORF2的蛋白則主要侷限於細胞質內;相較於ORF2蛋白的表現,ORF1蛋白於細胞內的表現與TUNEL陽性訊號呈現密切的相關性。本實驗的結果顯示PBL對於PCV2是具有感受性的,並且PCV2傾向於在分裂中的淋巴球內複製。根據目前所得的實驗結果,我們更進一步推測PCV2導致淋巴球凋亡與PCV2在其內複製可能具某種相關性,這些證據可以對PMWS的致病機制提供一部份的解釋。 | zh_TW |
dc.description.abstract | The porcine circovirus type Ⅱ (PCV2)-associated newly emerged disease, postweaning multisystemic wasting syndrome (PMWS), is characterized by lymphoid depletion in lymphoid organs and replacement by macrophages. Until now, whether PCV2 replicates in immune system and the pathogenesis of PMWS are still not well understood. In this study, we established a model of PCV2 infection in mitogen-treated peripheral blood lymphocytes (PBL). In the first part of the study, it was demonstrated that there was only 0.1% of PCV2 antigen-containing rate in PBL isolated from PCV2-free pigs and treated simultaneously with mitogen and PCV2. The results of both MTT assay and blastogenesis indicated that PCV2 could reduce the responsiveness of PBL to mitogen stimulation, but the results of survival and apoptotic rates indicated that PCV2 had no effects on the viability of PBL. In the second part of study, it was found PCV2 nucleic acid and antigens could be more easily detected in mitogen-stimulated PBL from clinically healthy PCV2-carrier pigs by in situ hybridization and immunofluorescence analysis, respectively. After 4 days of mitogen treatment, the PCV2 antigen-containing rate in PBL could reach 20% with IgM-positive cells predominant. The mitogen-stimulated PBL had higher PCV2 titer. Additionally, PCV2 ORF1-associated protein was usually present perinuclearly or within the nuclei; ORF2-associated protein was chiefly restricted in the cytoplasm; the expression of ORF1-associated protein, but not ORF2, as well matched with TUNEL signal. Results of this study indicate that PBL are indeed susceptible to PCV2 infection and PCV2 is capable to replicate in dividing lymphocytes. This replication may play a role on PCV2-induced apoptosis in PBL. The result may explain in part the pathogenesis of PMWS. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T05:47:53Z (GMT). No. of bitstreams: 1 ntu-95-R93629001-1.pdf: 3498272 bytes, checksum: 97d06cd0c3b8e49cc7f1fc41960c6973 (MD5) Previous issue date: 2006 | en |
dc.description.tableofcontents | 目錄
中文摘要 ………………………………………………………………….. I 英文摘要 ………………………………………………………………….. II 目錄 ………………………………………………………………….. III 表次 ………………………………………………………………….. VI 圖次 ………………………………………………………………….. VI 簡寫對照表 ………………………………………………………………….. VIII 第一章 序言 1 第二章 文獻回顧 3 第一節 豬環狀病毒 3 1-1 環狀病毒的病毒學分類 3 1-2 豬環狀病毒的結構組成 3 1-3 豬環狀病毒的感染以及複製模式 4 1-4 豬環狀病毒感染之標的細胞與組織內分佈 6 第二節 豬隻離乳後系統性耗弱症候群 8 2-1 歷史背景與流行病學 8 2-2 臨床症狀與病理變化 8 2-3 淋巴流失 10 2-3.1 淋巴組織內細胞組成之改變 10 2-3.2 周邊血液中細胞組成之改變 11 2-4 致病因子 12 2.4-1 其他病原的共同感染 12 2.4-2 免疫刺激或抑制 14 2.4-3 個體免疫的差異以及中和抗體的保護 15 2-5 其他PCV2相關的疾病或症候群 16 第三節 豬隻淋巴球對致裂原之反應 17 第三章 材料與方法 19 第一節 實驗設計 19 第二節 實驗材料 21 第三節 實驗方法 24 3-1 細胞的分離以及培養模式之建立 24 3-1.1 豬周邊血液淋巴球的分離與純化 24 3-1.2 致裂原刺激與病毒感作模式之建立 24 3-1.3 形態學觀察與樣本收取 25 3-1.4 trypan blue色素排除試驗 25 3-2 免疫螢光染色 25 3-2.1 細胞表面抗原染色 25 3-2.2 以螢光顯微鏡偵測細胞內抗原 26 3-2.3 以流式細胞儀偵測細胞內抗原 26 3-3 細胞凋亡之偵測 27 3-3.1 染色體螢光染料 27 3-3.2 TUNEL assay 28 3-3.3 Annexin V/ propidium iodide雙重染色 28 3-4 螢光顯微鏡與流式細胞儀的使用 29 3-5 細胞活性 30 3-5.1 細胞代謝能力MTT assay 30 3-5.2 細胞增生能力 Blastogenesis 30 3-6 原位雜交 31 3-7 病毒力價之檢測 31 第四章 結果 32 第一節 以無PCV2帶原、健康豬隻周邊血液淋巴球建立致裂原刺激與PCV2人工感染模式 32 4-1.1 豬周邊血液淋巴球的純化與對致裂原的反應 32 4-1.2 豬周邊血液淋巴球經致裂原刺激後的形態轉變 33 4-1.3 PCV2病毒感染的成立以及抗原表現量 33 4-1.4 細胞的存活率以及凋亡率 34 4-1.5 細胞活性 35 第二節 以PCV2自然帶原、健康豬隻周邊血液淋巴球進行致裂原刺激以及PCV2自然感染模式 42 4-2.1 PCV2抗原表現量 42 4-2.2 PCV2病毒抗原陽性細胞的形態學 43 4-2.3 PCV2病毒抗原陽性細胞所帶的表面抗原 44 4-2.4 細胞內PCV2 ORF1、ORF2的表現位置以及細胞凋亡之相關性 44 4-2.5 病毒核酸之偵測 46 4-2.6 病毒力價之檢測 46 第五章 討論 58 豬隻的淋巴球對PCV2具感受性 59 PCV2可在淋巴球內複製且其複製機制與淋巴球的細胞週期具顯著相關性 63 PCV2在淋巴球內大量複製的同時亦導致宿主細胞的死亡 67 第六章 參考文獻 72 | |
dc.language.iso | zh-TW | |
dc.title | 豬隻淋巴球對第二型豬環狀病毒感受性之探討 | zh_TW |
dc.title | The Susceptibility of Swine Lymphocytes to Porcine Circovirus Type Ⅱ | en |
dc.type | Thesis | |
dc.date.schoolyear | 94-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 鄭謙仁(Chian-Ren Jeng) | |
dc.contributor.oralexamcommittee | 鄭益謙(I-Chian Jeng) | |
dc.subject.keyword | 淋巴球,豬隻第二型環狀病毒, | zh_TW |
dc.subject.keyword | lymphocyte,porcine circovirus type 2, | en |
dc.relation.page | 81 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2006-07-11 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 獸醫學研究所 | zh_TW |
顯示於系所單位: | 獸醫學系 |
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