猪树状突细胞和肺泡巨噬细胞受猪生殖与呼吸综合症病毒感染後其类.doc

豬樹狀突細胞和肺泡巨噬細胞受豬生殖與呼吸綜合症病毒感 染後其類鐸接受體及細胞激素mRNA之表現 Expression Alteration of Cytokines mRNA in Porcine Dendritic Cells and Alveolar Macrophages in Response to Porcine Reproductive and Respiratory Syndrome Virus Infection 研究生：謝秉倫Hsieh, Bing-Lun 指導教授：鍾文彬Chung, Wen-Bin 【摘要】 樹狀突細胞 Dendritic cell；DC 和肺泡巨噬細胞 Alveolar macrophage；AM 為重要的抗原呈獻細胞，其上的pattern recognition receptors 如類鐸接受體 Toll-like receptor；TLR 與抗原分子結合後活化DC和AM並分泌細胞激素 Cytokines ，這些細胞激素可誘導T細胞發展為適當的作用細胞。本實驗以豬生殖與呼吸綜合症病毒 Porcine reproductive and respiratory syndrome virus；PRRSV 感染豬樹狀突細胞及豬肺泡巨噬細胞，於感染後不同時間萃取其mRNA，以RT-PCR或 real-time PCR測定TLRs，促發炎細胞素，趨化激素，Th1和Th2細胞激素mRNA含量。結果顯示，PRRSV感染後6小時DC和AM之TLRs mRNA表現量呈現短暫下降但未達顯著水準 P 0.05 ；受PRRSV感染之DC 其IL-1β、IL-8、IL-12p40、TNF-α及MCP-1的mRNA表現量呈現隨感染時間之增加而上升但只有IL-1β及TNF-α達顯著水準 P 0.05 ，AM受感染後其IL-1β、TNF-α、MCP-1及IFN-γ的mRNA表現量呈現隨感染時間之增加而上升但未達顯著水準 P 0.05 。 real-time PCR與RT-PCR測定的結果除了PRRSV感染AM之IL-10和TLR3 mRNA之定量兩種方法所得結果之相關係數 r 0.12和r 0.06 較低外，其它結果成高度相關，其相關係數達到0.62-0.8。鍵詞：樹狀突細胞、肺泡巨噬細胞、豬生殖與呼吸綜合症病毒、類鐸接受體和細胞激素 Dendritic cell DC and Alveolar macrophage AM are important antigen-presenting cells. The binding between pattern recognition receptors such as Toll-like receptors TLR on DC and AM and pathogen-associated molecular pattern resulted in the secretion of various cytokines which the modulate T cell responses of appropriate effector phenotypes accordingly. In this study, porcine immature DC imDC and AM were infected with porcine reproductive and respiratory syndrome virus PRRSV . The mRNA expressions of TLRs, pro-inflammatory cytokines, chemokines, T help 1 Th1 and Th2 cytokines were detected by reverse transcription-polymerase chain reaction RT-PCR and real-time PCR from cells at various periods of times after infection. Results showed that the mRNA expression of TLRs in DCs and AMs had a transient and insignificant decrease 6 hours after the infection. Thereafter, the mRNA expression of IL-1β, IL-8, IL-12p40, TNF-α and MCP-1 in