在HTLV-1感染的T细胞中HMGB1、Tax蛋白与细胞自噬关系的研究

近来人们发现高迁移率族蛋白1（HMGB1）是一个新的Beclin 1结合蛋白参与细胞自噬的调节。但HMGB1对HTLV-1病毒感染细胞内的自噬是如何调节的？HTLV-1病毒的复制与细胞自噬的关系如何？目前尚不清楚。我们拟利用透射电子显微镜、共聚焦显微镜和流式细胞仪等仪器，采用免疫荧光染色、免疫共沉淀、荧光素酶报告基因、RNAi沉默和Western blot等方法，从分子和细胞水平对HMGB1、Tax蛋白和细胞自噬的关系进行研究，系统地观察细胞外和细胞内的HMGB1对HTLV-1病毒的复制和细胞自噬的影响。本项目将首次揭示HMGB1对HTLV-1病毒感染细胞中细胞自噬的作用机制，证实HMGB1在HTLV-1感染细胞的病毒复制和成人T细胞白血病（ATL）的发生中扮演重要角色，为将HMGB1作为阻止HTLV-1病毒复制和ATL发生的重要基因靶点，建立起有效的基因治疗方法奠定基础。

Recently, it has been shown that HMGB1 is a novel Beclin 1-binding protein and could regulate autophagy. But they are unclear that HMGB1 how to regulate autophagy in HTLV-1 infected cells, and the relationship of HTLV-1 viral replication and autophagy. We will investigate the relationship of HMGB1, Tax protein and autophagy from molecular and cellular levels，and observe systematically the effect of extracellular and intracellular HMGB1 on HTLV-1 Viral Replication and autophagy by the methods of immune fluorescence staining, co-Immunoprecipitation, luciferase reporter gene, RNAi silence and western blot etc., and using the instrument of transmission electron microscope, confocal microscope and flow cytometry etc. It will be firstly discovered that the mechanism of HMGB1 action to autophagy, and confirmed that HMGB1 play an important part in viral replication and causing adult T-cell leukemia in HTLV-1 infected cells after finishing the project. The results will lay the foundation of HMGB1 as the target of gene therapy to inhibit HTLV-1 viral replication and ATL occurrence.