TRIM家族蛋白：TRIM22、TRIM14抑制流感病毒的功能和机制的初步研究

发布时间：2018-06-17 20:15

本文选题：TRIM22 + TRIM14　；参考：《北京协和医学院》2016年硕士论文

【摘要】：流感病毒(influenza virus)是一种流行广泛、传染性极强的负链病毒,可在高危人群中造成严重疾病和死亡。每年流感在世界范围内的流行造成约300万至500万例严重疾病和约25万至50万例死亡。流感病毒分为A、B、C三型,其中A型流感病毒(influenza A virus, IAV)致病性最强。目前治疗流感的药物有NA的抑制剂扎那米韦、奥司米韦,及针对M2的抑制剂金刚烷胺,但是流感病毒通常会对药物产生耐药性。虽然疫苗能够预防病毒感染,但流感病毒每年流行的毒株都不一样而且病毒的突变性很强容易导致疫苗错配,单纯靠疫苗预防感染并不现实。因此,有必要寻找新的抑制流感病毒的机制,为新药物的研发提供线索。在固有免疫对抗流感病毒感染过程中干扰素发挥了重要作用。干扰素能够诱导表达ISG(interferon-stimulated gene)。ISG通过改变细胞的代谢和内环境来调节机体免疫反应或者与病毒的某些组分发生直接的相互作用来抑制病毒的复制。这些ISG中包括较多的TRIM蛋白,TRIM蛋白家族是由N端到C端以RING、B-box、 Coiled-coil结构域依次排列为结构特征的一类蛋白,TRIM14和TRIM22属于TRIM (tripartite motif)家族成员。目前已经发现一些TRIM蛋白能够抑制HBV、HIV、 IAV等多种病毒,但TRIM14与流感病毒的关系还不清楚。本研究以TRIM22为研究对象来验证BiLC系统能否快速有效的检测ISGs蛋白与流感蛋白相互作用；同时建立了稳定表达细胞系的构建方法及快速检测流感病毒复制的系统。与文献报道一致,TRIM22能有效抑制流感病毒的复制,并且BiLC系统检测到TRIM22与NP蛋白有特异的结合。基于上述实验方法,我们研究了TRIM14与流感病毒的关系。发现TRIM14属于ISG并且主要被I型干扰素诱导高表达。过表达TRIM14和敲除内源TRIM14细胞的病毒扩增实验都证明TRIM14对流感病毒具有抑制作用。利用高效BiLC系统检测发现TRIM 14可能通过与M2、NP、PA、NS1等蛋白相互作用来抑制流感病毒复制。上述研究的完成,为快速研究ISG与流感病毒的关系和机制提供了新的方法。丰富了固有免疫对流感病毒抑制方式的认识,同时也为研究抗流感病毒药物提供了新的视角。
[Abstract]:Influenza virus Influenza virus (Influenza virus) is a widespread and highly infectious negative chain virus, which can cause serious disease and death in high risk population. A worldwide pandemic causes about 3 million to 5 million serious diseases and 250000 to 500000 deaths each year. Influenza virus can be classified into three types, type A influenza virus influenza A virus (IAV) is the most pathogenic. Current treatments for influenza include anamivir, an inhibitor of na, osmevir, and amantadine, an inhibitor of M2, but influenza viruses are often resistant to drugs. Although the vaccine can prevent the infection of the virus, the influenza virus each year is different, and the mutation of the virus is very easy to cause the vaccine mismatch, it is not realistic to rely on the vaccine to prevent the infection. Therefore, it is necessary to find new mechanisms to inhibit influenza virus and provide clues for the development of new drugs. Interferon plays an important role in innate immunity against influenza virus infection. Interferon can induce the expression of ISG interferon-stimulated gene. ISG inhibits the replication of the virus by changing the metabolism and internal environment of the cells to regulate the immune response or to interact directly with some components of the virus. These ISGs include more trim protein family, which are members of tritrite motif family from N-terminal to C-terminal with RING-B-box. Coiled-coil domain is arranged in order of structural characteristics. Some trim proteins have been found to inhibit HIV, IAV and other viruses, but the relationship between TRIM14 and influenza virus is not clear. In this study, TRIM22 was used to verify the ability of BiLC system to detect the interaction between ISGs protein and influenza protein, and to establish a stable expression cell line and a system for rapid detection of influenza virus replication. As reported in the literature, TRIM22 can effectively inhibit the replication of influenza virus, and the specific binding of TRIM22 to NP protein was detected by BiLC system. Based on the above experimental method, we studied the relationship between TRIM14 and influenza virus. It was found that TRIM14 belongs to ISG and is mainly induced by type I interferon. Both overexpression of TRIM14 and knockout of endogenous TRIM14 cells showed that TRIM14 had inhibitory effect on influenza virus. Highly efficient BiLC detection revealed that TRIM14 may inhibit influenza virus replication by interacting with proteins such as M2, NPN, PANS1 and so on. The above research provides a new method to study the relationship and mechanism between ISG and influenza virus. It not only enriches the understanding of innate immunity to the inhibition of influenza virus, but also provides a new perspective for the study of anti-influenza drugs.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R373.13

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