TRIM22调控人巨细胞病毒复制的作用及机制

Human cytomegalovirus (HCMV) is one of the main pathogens of perinatal infection, which seriously threatens the quality of population in china. But the mechanism of HCMV infection has not been fully defined to limit the prevention and treatment of HCMV infection. Although studies have shown that TRIM22 can regulate the replication of various viruses, the relationship between TRIM22 and HCMV infection is not clarified. Our previous study showed that TRIM22 was significantly elevated in THP-1 cells infected with HCMV. On this basis, firstly we will explore the regulatory role of TRIM22 in HCMV replication and the time of the regulation by controlling the expression of TRIM22 in THP1 cells infected with HCMV. Secondly we will identify HCMV proteins or DNA sequences which are combined with TRIM22 by using the co-immunoprecipitation method. Finally we will reveal the key structure of TRIM22 inhibiting HCMV replication by over-expressing the knockout RING domain or SPRY structure of TRIM22 in THP 1 cells infected with HCMV. Eventually the mechanism of TRIM22 regulating HCMV replication will be revealed through the study, and we will also provide a new idea about the relationship between HCMV infection and host immunity.

人巨细胞病毒（HCMV）是围产期感染主要病原体之一,严重威胁我国人口质量，而HCMV感染机制尚未完全明确限制了临床HCMV的防治。研究表明TRIM22对病毒感染具有调节作用，但TRIM22与HCMV感染之间的关系尚待阐明。我们前期实验发现，HCMV感染THP-1细胞后，TRIM22的表达水平显著升高。本项目拟在此基础上，首先在HCMV感染的THP1细胞中调控TRIM22的表达水平，探索TRIM22对HCMV复制的调控作用及调控时间点；然后对TRIM22的结合蛋白和DNA序列进行免疫共沉淀，鉴定TRIM22结合的HCMV蛋白或DNA序列；最后在HCMV感染的THP-1细胞中过表达敲除RING结构域或SPRY结构域的TRIM22，揭示TRIM22抑制HCMV复制的关键结构域。通过上述研究揭示TRIM22对HCMV复制水平调控的作用及机制，将为深入探索HCMV感染与宿主免疫的关系提供新思路。

人巨细胞病毒(Human cytomegalovirus, HCMV)是围产期感染的主要病原体之一，易导致严重后果。HCMV的复制增殖是HCMV感染致病的关键环节。本研究前期发现在PMA诱导分化的THP-1巨噬细胞中感染HCMV可以促进抗病毒蛋白TRIM22的表达, 且表达水平随着时间和病毒滴度的增加而增加。在此基础上，我们进一步验证了TRIM22表达水平的增加是IFN-β所介导的。同时，HCMV感染THP-1巨噬细胞也诱导了IL-6和TNF-α表达水平的增加。在HCMV感染的THP-1巨噬细胞中，下调TRIM22蛋白表达后，HCMV复制水平显著增加。进一步在HCMV感染的THP-1巨噬细胞中过表达TRIM22蛋白、缺失RING结构域的TRIM22蛋白或缺失SPRY结构域的TRIM22蛋白，HCMV在过表达TRIM22蛋白组复制水平被显著抑制，其它组无差异。最后，免疫共沉淀TRIM22的结合蛋白并进行质谱分析，提示TRIM22蛋白可能和UL84蛋白结合而抑制病毒复制。本课题通过上述研究阐明了HCMV感染THP-1巨噬细胞后，通过IFN-β介导了抗病毒蛋白TRIM22表达水平的增加，TRIM22进一步抑制HCMV的复制的具体机制。本研究初步明确了TRIM22与HCMV 感染之间的具体调控机制，一方面可为今后进一步阐明HCMV的致病机制提供新思路，另一方面也为HCMV 增殖感染的防治提供了潜在靶点。