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題名 | Gene Expression Profiling of the Liver in Chimpanzees during HCV Infection Using Microarray: Genes Related to Viral Persistence and Clearance=利用微陣列探討黑猩猩受C型肝炎病毒感染後的肝臟基因表達圖譜:病毒續存與清除的相關基因 |
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作 者 | 胡志棠; | 書刊名 | 慈濟醫學 |
卷期 | 17:3 民94.06 |
頁次 | 頁145-154+200 |
分類號 | 415.5332 |
關鍵詞 | C型肝炎; 基因表達圖譜; 黑猩猩; Hepatitis C; Gene expression profiling; Chimpanzees; |
語文 | 英文(English) |
中文摘要 | 目的:在於比較黑猩猩受C型肝炎病毒(HCV)感染後,追蹤其病毒持續存在或已被排除後的肝臟基因表達圖譜的變化。另外,藉黑猩猩感染後此不同的預後,試圖找出與HCV感染性、病毒清除、或病毒持續存在相關的基因群。材料與方法:在美國西南基金會生物醫學研究中心(Southwest Foundation for Biomedical Research)四隻黑猩猩接受肝臟HCV(基因型1b)的接種或由血清感染的途徑引發肝炎的產生。黑猩猩每週接受抽血及定期肝臟組織切片檢查,並定期測是其病毒血症的濃度、肝功能指數、判讀肝臟組織發炎程度、及肝臟組織中干擾素-γ、干擾素-α、和interleukin-4的分泌濃度。肝臟組織切片中其RNA藉Trizol萃取後用RT-PCR擴增。一個cDNA選殖基因庫PCR擴增其內的cDNA後,利用基因微陣列技術在玻片上鑲點出8500點不同的基因以探討黑猩猩受C型肝炎病毒感染後肝臟基因表達濃度的變化。結果:九個與第一型干擾素反應相關的基因在受感染後能康復的黑猩猩X0190於感染早期即明顯地表達增加,此情況在變為慢性化感染的黑猩猩(X0142及X0234)也有相同的現象。十四個與生長/訊號傳遞相關的基因在黑猩猩X0190受感染後都在早期即明顯地表增加,但大部分這些基因的表達在變為慢性化感染的黑猩猩(X0142及X0234)中卻沒有增加。於HCV病毒被清除期間及之後,黑猩猩X0190受感染後導致與細胞免疫力相關的基因表達增加,此與其免疫系統能引發足夠的CD8+T細胞反應以清除病毒相吻合,但此等基因的表達在變為慢性化感染的黑猩猩(X0142及X0234)卻沒有增加的現象。黑猩猩X0132在各種HCV病毒接種後,在不增加與第一型干擾素反應與細胞免疫力相關的基因表達的情況下,都能限制和清除病毒感染。結論:十五個與生長/訊號傳遞相關的基因中,其中至少六個基因僅於HCV急性、非持續性感染時表達明顯增加,可作為早期判斷HCV感染後病毒將可被清除或變成慢性肝炎的重要指標。至於黑猩猩X0132如何在沒有引發明顯先天性和適應性免疫反應下,仍能阻止並清除HCV感染,其機轉值得深入探討。 |
英文摘要 | Objectives: This study characterized and compared hepatic gene expression profiles during hepatitis C virus (HCV) infection in which viremia persists or is eliminated. In addition, we identified genes associated with infectivity, clearance and persistence. Materials and Methods: Four chimps housed at the Southwest Foundation for Biomedical Research were inoculated with transcripts from a HCV genotype 1b molecular clone or monotypic serum derived from the infection. Animals were bled weekly and needle punch biopsies were collected at the indicated time points. Assays were performed to measure viremia, alanine transaminase (ALT) levels, biopsy histology, and intrahepatic cytokine levels (IFN-gamma, IFN-alpha, and IL-4). RNA was collected from frozen biopsy materials using Tizol and was then amplified. An 8500-spot cDNA microarray print was generated by mechanically applying PCR products from a library of cDNA clones. Results: Of the 9interferon type 1 response genes significantly induced early during infection in chimp X0190 which acquired acute infection with recovery, most were also significantly induced at some point in chimps that became chronically infected (X0142 and X0234). Most of the 14 growth/signal transduction genes significantly induced early during infection in chimp X0190 were not induced in the chronically infected chimps X0142 and X0234. Significantly induced genes involved in cell-mediated immunity during and after viral clearance in chimp X0190 were consistent with a CD8+ T-cell response, which was not so universally evident and outcome-specific in chimps X0142 and X0234. Chimp X0132 was able to limit and clear the infection without induction of these cell-mediated immunity and interferon type 1 response genes. Conclusions: At least 6 out of 15 of the growth/signal transduction genes highly induced in acute but not chronic infection can be used as potential markers for predicting subsequent viral clearance or persistence after infection. The mechanism for chimp X0132 being able to limit and clear different strains of HCV without induction or robust innate and adaptive immune responses remain elusive. |
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