大豆来源的蛋白酶抑制剂（BBI和KI）抑制HIV感染巨噬细胞

发布时间：2018-03-11 09:54

本文选题：巨噬细胞　切入点：BBI　出处：《武汉大学》2016年博士论文　论文类型：学位论文

【摘要】：大豆含有多种具有生物活性的蛋白质和多肽,Kunitz抑制剂(KI)和Bowman-Birk抑制剂(BBI)是大豆中最丰富的两种蛋白酶抑制剂,具有显著的抗炎作用。BBI和KI的抗炎作用已经在体外实验、动物模型和临床研究中得到了充分的证实。炎症可导致免疫激活,而免疫激活和炎症与HIV感染者的疾病进程相关。宿主天然免疫反应在控制炎症和病毒感染中发挥了重要作用。巨噬细胞是天然免疫系统的主要免疫细胞。巨噬细胞在HIV感染的发病机制中的重要性体现在其双重作用：一方面它参与宿主的抗HIV免疫反应；另一方面它是HIV的主要靶细胞和潜藏库。大量研究表明,HIV能抑制宿主细胞的抗病毒信号通路,导致慢性持续感染。由于HIV可整合于宿主细胞的DNA内,清除靶细胞中的病毒是治愈HIV感染的关键。因此,激活被HIV抑制的靶细胞内的抗病毒免疫反应有利于控制和清除病毒。1.BBI抑制HIV进入巨噬细胞BBI是一种水溶性的小分子蛋白质。作为大豆来源的蛋白酶抑制剂,BBI具有抗炎作用。早期的研究表明,BBI在非细胞体系中可抑制HIV的逆转录酶活性。但目前尚无有关BBI在细胞水平抑制HIV感染的报道。在本研究中,我们探讨了BBI对HIV进入细胞的影响。结果显示,BBI可抑制HIV进入巨噬细胞。对其机制的研究表明,BBI可降低HIV进入细胞的主要受体CD4和辅助受体CCR5的表达,此外,BBI还能增加巨噬细胞产生CCR5的配体,CC趋化因子(RANTES,MIP-1α和MIP-1β)的表达。CC趋化因子的中和抗体可抵消BBI介导的抗HIV效应。这些结果表明,BBI具有阻止HIV进入靶细胞的功能。2.BBI和KI通过JAK/STAT信号通路抑制HIV感染巨噬细胞巨噬细胞在炎症和免疫激活中发挥着重要作用,而免疫激活和炎症与HIV感染者的疾病进程密切相关。作为大豆来源的胰蛋白酶和胰凝乳蛋白酶抑制剂,BBI和KI具有显著的抗炎作用。在本研究中,我们探讨了BBI和KI对HIV感染巨噬细胞的影响。我们的研究显示,BBI和KI能有效地抑制HIV感染巨噬细胞,并且没有细胞毒作用。对其机制的研究显示,BBI和KI处理的巨噬细胞能产生多种抗病毒ISGs,包括Mx2,OAS-1,viperin,ISG15和ISG56.BBI和KI还能诱导HIV限制因子的表达,包括APOBEC3F,APOBEC3G和tetherin.除此之外,BBI和KI处理的巨噬细胞可诱导IFN-β的表达。IFN-β是JAK/STAT信号通路的主要激活剂。用JAK抑制剂阻断JAK/STAT信号通路可抵消BBI和KI诱导的抗病毒因子和抗HIV效应。这些结果说明,BBI和KI可激活巨噬细胞内的天然免疫反应,产生多种能在HIV复制周期的多个关键步骤发挥抑制作用的抗病毒因子。因此,BBI和KI具有预防和治疗HIV感染的潜在药用价值。
[Abstract]:Soybean contains a variety of bioactive protein and polypeptide Kunitz inhibitor (Ki) and Bowman-Birk inhibitor (BBI) are two of the most abundant protease inhibitors in soybean. The anti-inflammatory effects of BBI and Ki have been tested in vitro. Animal models and clinical studies have been well documented. Inflammation can lead to immune activation, Host innate immune response plays an important role in controlling inflammation and viral infection. Macrophages are the main immune cells in the innate immune system. The importance of cell in the pathogenesis of HIV infection lies in its dual role: on the one hand, it participates in the host's anti-#en1# immune response; On the other hand, it is the main target cell and latent library of HIV. A large number of studies have shown that HIV can inhibit the antiviral signaling pathway of host cells and lead to chronic persistent infection. Because HIV can be integrated into DNA of host cells, Removing viruses from target cells is the key to curing HIV infection. Activation of antiviral immune response in target cells inhibited by HIV is beneficial to the control and clearance of virus. 1. BBI inhibits the entry of HIV into macrophages BBI is a water-soluble small molecular protein. Antiinflammatory effects. Early studies have shown that BBI can inhibit the reverse transcriptase activity of HIV in non-cellular systems. However, there is no report of BBI inhibiting HIV infection at the cell level. We investigated the effect of BBI on the entry of HIV into macrophages. The results showed that BBI could inhibit the entry of HIV into macrophages. The study of its mechanism showed that BBI could reduce the expression of CD4 and CCR5, the main receptor and coreceptor of HIV into the cells. In addition, BBI could also increase the expression of CCCC chemokines (RANTESMIP-1 伪 and MIP-1 尾) in macrophages. The neutralizing antibodies of CC chemokines could counteract the anti-#en3# effect mediated by BBI. These results suggest that BBI can prevent HIV from entering target cells. 2.BBI and Ki play an important role in inflammation and immune activation by inhibiting macrophages infected with HIV through JAK/STAT signaling pathway. However, immune activation and inflammation are closely related to the disease progression of HIV infection. Trypsin and chymotrypsin inhibitors, such as trypsin and chymotrypsin, have significant anti-inflammatory effects in this study. We investigated the effects of BBI and Ki on HIV infected macrophages. Our results showed that BBI and Ki could effectively inhibit HIV infected macrophages. The study of its mechanism showed that macrophages treated with BBI and Ki could produce many kinds of anti-virus ISGs, including Mx2OAS-1 viperinine ISG15, ISG56.BBI and Ki, which could also induce the expression of HIV limiting factors. Besides, macrophages treated with BBI and Ki can induce IFN- 尾 expression. IFN- 尾 is the main activator of JAK/STAT signaling pathway. Blocking JAK/STAT signaling pathway with JAK inhibitor can counteract anti-viral factor and anti-#en4# effect induced by BBI and Ki. These results suggest that BBI and Ki can activate innate immune responses in macrophages, It produces a variety of antiviral factors that can inhibit HIV infection in several key steps of HIV replication cycle. Therefore, BBI and Ki have potential medicinal value in the prevention and treatment of HIV infection.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R512.91

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