2011-05-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656671摘要：登革出血能導因於一次感染，但在不同血清型病毒所引起的二次感染時，有很高的風險會產生登革出血熱及登革休克症。一次感染所產生之非中和性抗體具有增強第二次感染之病毒進入細胞以及記憶性T 細胞產生TNF 是目前用來解釋二次感染風險的兩個假說。之前建立的一次感染出血的動物模式中，我們發現登革病毒會感染內皮細胞並吸引產生大量的TNF 的巨噬細胞聚集於血管周圍。我們更進一步建立二次感染登革出血的小鼠模式，小鼠接受第一型登革病毒後再給予第二型登革病毒，以較低的病毒量就可引發小鼠出血，除了巨噬細胞，會產生TNF 的CD4 和CD8 T 細胞也參與產生出血的過程。目前此論文正在撰寫中。在這一年期延續性計畫中，我們將持續並更深入的研究原計畫的目標二及目標三。目標二，我們將分析登革病毒感染後，內皮細胞及免疫細胞上黏著分子的變化，探討產生不同細胞激素的免疫細胞所表現之不同黏著分子如何影響免疫細胞在組織中的移動，並且研究以不同型或同型病毒胜肽刺激T 細胞是否產生有致病性或保護性的細胞激素。目標三，研究登革病毒NS2B-protease 如何調控內皮細胞的功能與命運。我們發現並已確認與NS2B-protease 結合的細胞蛋白INP-1，接下來會探討INP-1 跟登革病毒的NS2B-protease 作用之分子機制與對內皮細胞功能與命運之影響。<br> Abstract: Dengue hemorrhage can result from primary infection, but there is a greater risk to developDHF/DSS during secondary infection by a heterotypic DV. Non-neutralizing antibody fromprimary infection enhancing the uptake of the second infecting DENV and activation ofcross-reactive memory T cells and cytokine production are two most prevalent hypotheses toexplain the risk of secondary infection.In the dengue hemorrhage mouse model of primary infection we showed that both DVtargeting endothelial cells and TNF production by the infiltrating macrophages in the vicinityof endothelium are critical to hemorrhage development. As proposed in the original grantproposal, we further created a hemorrhage mouse model of secondary infection. Micereceiving sequential infections of DV-1 and DV-2 developed severe/systemic hemorrhage atmuch lower virus titers. Besides macrophages, both TNF-producing CD4+ and CD8+ Tcells were involved in hemorrhage development. This model offers an unprecedentedopportunity to study the immunopathogenic mechanisms of dengue hemorrhage aftersecondary infection by a heterotypic DV. A manuscript describing these findings is inpreparation for publication.In this one-year continuation proposal, we will continue the work proposed inSpecific Aims 2 and 3 of the original proposal. In Specific Aim 2, we will study thedifferential molecular changes and cross-reactivity relating to the migration of protective vs.pathogenic immune cells. Utilizing the animal model, we will be able to study the interplayamong DV, activated immune cells and endothelium in situ where the drama of the interplaytakes place. We will also analyze whether T cells reacting to homotypic vs. hterotypicpeptides produce pathogenic or protective cytokines and whether IFN-γ is protective indengue infection. In Specific Aim 3, we will investigate the interaction between DVNS2B-protease and endothelial cells that alters endothelial cell functions and cell fate. Wehave identified and confirmed the cellular interacting protein (dubbed INP-1) ofNS2B-protease. We will be devoted to investigate the molecular interactions betweenINP-1 and NS2B-protease and assess the effects of the interaction on endothelial cells.