2011-01-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/648977摘要：CD4+ T 淋巴細胞與巨嗜細胞(Macrophage)是愛滋病毒(HIV)主要感染的標的細胞。愛滋病毒感染巨噬細胞時會導致低量病毒複製與持續性感染現象(persistent infection)，但受感染的CD4+ T 淋巴細胞則會導致細胞增生(clonal expansion)、大量複製病毒、繼而產生細胞凋亡(apoptosis)。CD4+ T 淋巴細胞與巨嗜細胞對愛滋病毒[易感染性](cell susceptibility)的差異性在科學的領域一直是重要的研究議題。我們實驗室發現DnaJ 伴護蛋白家族(DnaJ chaperon protein family)的兩種同型異構蛋白(isoform protein): DNAJB6 與MRJ是可以與愛滋病毒type 1 (HIV-1)Vpr蛋白交互作用的細胞蛋白。初步的西方墨點實驗結果顯示DNAJB6 與MRJ蛋白均可大量表現在活化的周邊淋巴細胞(PBL)，但周邊單核球培養出來的巨嗜細胞可表現同等量MRJ蛋白，卻表現極低量的DNAJB6。相關研究文獻指出DNAJB6、大型的MRJ 同型異構蛋白，是愛滋病毒type 2 (HIV-2) Vpx蛋白的作用蛋白，它的功能是促進HIV-2 PIC (pre-intergration complex)的核運送(nuclear import)，幫助病毒複製。在這個三年計畫中，我們想釐清究竟DNAJB6 與MRJ這兩種同型異構伴護蛋白在不同類型的細胞上對愛滋病毒的易感性上扮演的角色與功能是什麼。我們預定在CD4+ T細胞株與巨嗜細胞株建立DNAJB6 與MRJ 降低( knock-down )或過量表現(over-expression)的穩定系統(stable clones)，再來檢視分析愛滋病毒如何感染這些細胞株及後續的複製現象。其它與DNAJB6 與MRJ相關參與在病毒感染、複製(replication)等不同步驟的分子調控機轉，也將在這個計畫內陸續被研究釐清。我們相信此研究計畫可以幫忙了解不同類型的細胞對愛滋病毒感染的差異性，與引領開啟不同伴護蛋白可能在細胞[易感性]扮演不同的重要生物功能的嶄新研究方向。<br> Abstract: CD4+ T cells and macrophage are main targets for HIV-1 infection. HIV-1 infection in macrophage is usually low grade and persistent, while infection of CD4 T cells leads to explosive productive infection and death of T cells. This different susceptibility of CD4+ T cells and macrophage to HIV-1 has been an important topic. We had identified two isoforms of DnaJ-related chaperon proteins, DNAJB6 and MRJ, to be interacting proteins of Vpr. Our preliminary data showed that high levels of DNAJB6 and MRJ were present in activated peripheral blood lymphocyte (PBL).On the contrast monocyte-derived macrophages (MDM) possessed equivalent levels of MRJ but low level of DNAJB6. One previous study demonstrated that DNAJB6, large isoform of MRJ, is an HIV-2 Vpx-interacting protein and could promote nuclear import of preintergration complex (PIC). In this 3 year coming project, we want to address whether the differential expression of DNAB6 and MRJ determines cell susceptibility to HIV-1 infection. We will set up DnaJB6 and MRJ knock-down or over-expressed stable clones both in CD4+ T cells and macrophage cell line and examine how HIV-1 replicate in these different cells. Detailed mechanism about how DNAJB6 and MRJ moderate HIV-1 infection will be further dissected in this project. We believe this project will shed the light on how different cells react to HIV-1 infection and demonstrate that chaperon proteins may play an important role in cellular susceptibility to some virus infection.愛滋病毒伴護蛋白易感染性HIV-1DNAJB6MRJcell susceptibility