Association of TRIM22 with the type 1 interferon response during primary human cytomegalovirus infection in THP-1 macrophages
Association of TRIM22 with the type 1 interferon response during primary human cytomegalovirus infection in THP-1 macrophages
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TRIM22 与 THP-1 巨噬细胞中原发性人巨细胞病毒感染期间 1 型干扰素反应的关联
DOI：
10.32604/biocell.2019.08177
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发表时间：
2019
期刊：
影响因子：
1.2
通讯作者：
Shang Shiqiang
中科院分区：
文献类型：
--
作者：
Li Wei;Gao Huihui;Tao Ran;Liu Lifang;Shang Shiqiang
作为干扰素的反应因子，三方基序(Trim)22被报道具有抗病毒活性。本研究以人巨细胞病毒(HCMV)感染THP-1巨噬细胞，建立人巨细胞病毒裂解感染模型。IL-6、肿瘤坏死因子-α(TN-α)和干扰素-β(β)在不同感染时间和不同滴度下均显著上调。此外，首次发现在巨细胞病毒感染过程中，THP-1巨噬细胞中TRIM22的表达在mRNA和蛋白水平上均上调。此外，干扰素-β还能诱导巨噬细胞表达TRIM22。缺失TRIM22后，随着HCMV滴度和蛋白含量的增加，HCMV复制活性增强。综上所述，首次报道了巨细胞病毒可通过干扰素-β信号途径诱导TRIM22激活，而TRIM22可抑制巨噬细胞中巨细胞病毒的复制。2019.08177人巨细胞病毒是一种人类巨细胞病毒，是一种具有双链基因组的β-疱疹病毒，在人类中广泛存在(程等，2017年)。在健康的成年人中，巨细胞病毒感染通常是无症状的；然而，在免疫功能低下的患者或先天性感染的婴儿中，感染通常要严重得多(Aiello等人，2019年)。人巨细胞病毒在其人类宿主中具有广泛的细胞嗜性(Sinzger等人，2008年)。在不同的细胞或器官中，HCMV感染可能激活直接或间接参与病毒复制的细胞因子(Clement和Humphreys，2019)。三方基序(TRIM)家族包含70多种人类蛋白质，参与细胞增殖、分化、凋亡、肿瘤发生和免疫等许多生物学过程(Nisole等人，2005年；Ozato等人，2008年)。TRIM成员是广泛和保守的蛋白质，其特征是三体结构包括环域(Lian等人，2017)。许多TRIM家族成员可被干扰素诱导，并已成为参与先天性免疫和获得性免疫的抗病毒分子(Hattlmann等人，2012年)。TRIM5α是逆转录病毒的有效限制因子，包括N嗜性小鼠白血病病毒和人类免疫缺陷病毒-1(Stremlau等人，2004年；Yap等人，2004年)。TRIM19具有抗水疱性口炎病毒、HIV1和甲型流感病毒的活性(Nisole等人，2005年)。TRIM79NS5α可以通过黄病毒NS5蛋白的降解来对抗森林脑炎病毒的复制(Taylor等人，2011年)。*地址对应：石强尚，shangsq@zju.edu.cn人类TRIM22基因位于第11号染色体上，与TRIM5、TRIM6和TRIM34基因相邻(Vicenzi和Poli，2018)。与其他TRIM蛋白一样，TRIM22有一个与E3连接酶有同源性的环状结构域。通过降解病毒周期的关键因素，E3连接酶活性对于TRIM22介导的抗病毒活性非常重要(Barr等人，2008年；Di等人，2013年；Uchil等人，2008年)。据报道，TRIM22参与了对抗脑心肌炎病毒(ECMV)、乙肝病毒、艾滋病毒-1和甲型流感病毒的研究(Barr等人，2008年；Di等人，2013年；Hattlmann等人，2012年)。本研究旨在探讨人巨细胞病毒在初次感染过程中对TRIM22表达的调节作用。材料与方法采用THP-1单核细胞培养方法，在含有10%胎牛血清(美国Gibco)、青霉素G(100U/m L)、硫酸链霉素(100μg/m L)、L-谷氨酰胺(1%)和非必需氨基酸(1%)的RPMI1640培养液中培养。细胞培养温度为37℃，二氧化碳浓度为5%。用100 ng/mL佛波酯(北京，索拉比奥，中国)处理THP-1单核细胞，以确定THP-1单核细胞已分化为THP-1巨噬细胞。用100μL裂解缓冲液(大安基因有限公司，中国)，反复冻融3次，收集实时荧光定量聚合酶链式反应THP-1巨噬细胞。混合物在100°C下煮沸10分钟。然后将混合物在13000 g的温度下离心5分钟。
As a response factor of interferon, tripartite motif (TRIM) 22 was reported to exert antiviral activity against viruses. In this study, THP-1 macrophages were infected with human cytomegalovirus (HCMV) to establish the HCMV lytic infection model. The mRNA levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and interferonbeta (IFN-β) were significantly up-regulated in THP-1 macrophages at different infection time and titers. Moreover, for the first time, upregulation of TRIM22 expression was found during HCMV infection at both mRNA and protein levels in THP-1 macrophages. Furthermore, IFN-β could induce TRIM22 expression in THP-1 macrophages or HCMV infected THP-1 macrophages. Depletion of TRIM22 increased replication activity of HCMV with increasing of HCMV titers and HCMV proteins. In conclusion, it is the first report that HCMV can induce TRIM22 activation through IFN-β signaling and TRIM22 can suppress replication of HCMV in THP-1 macrophages. BIOCELL 2019 43(4): 285-291 Doi: 10.32604/biocell.2019.08177 www.techscience.com/biocell Introuction Human cytomegalovirus (HCMV) is a beta-herpesvirus with the genome of double-stranded DNA, which is widespread in human populations (Cheng et al., 2017). HCMV infection is generally asymptomatic in healthy adults; however, infections are often much more severe in immunocompromised patients or congenitally infected infants (Aiello et al., 2019). HCMV has a wide cell tropism within its human host (Sinzger et al., 2008). In different cells or organs, HCMV infection may activate cytokines that are directly or indirectly involved in virus replication (Clement and Humphreys, 2019). The tripartite motif (TRIM) family contains more than 70 human proteins that are involved in many biological processes such as cell proliferation, differentiation, apoptosis, oncogenesis and immunity (Nisole et al., 2005; Ozato et al., 2008). TRIM members are wide and well-conserved proteins characterized by a tripartite structure comprising a RING domain (Lian et al., 2017). Many TRIM family members are inducible by interferon (IFN) and have emerged as antiviral molecules involved in both innate and adaptive immunity (Hattlmann et al., 2012). TRIM5α is a potent restriction factor of retroviruses, including N-tropic murine leukemia virus (NMuLV), and human immunodeficiency virus-1 (HIV-1) (Stremlau et al., 2004; Yap et al., 2004). TRIM19 has activity against vesicular stomatitis virus, HIV1, and influenza A virus (Nisole et al., 2005). TRIM79α can against tick-borne encephalitis virus replication through the flavivirus NS5 protein degradation (Taylor et al., 2011). *Address correspondence to: Shiqiang Shang, shangsq@zju.edu.cn The human TRIM22 gene lies on chromosome 11, which is adjacent to TRIM5, TRIM6, and TRIM34 genes (Vicenzi and Poli, 2018). Like other TRIM proteins, TRIM22 has a RING domain that has homology with E3 ligases. E3 ligase activity is important for TRIM22-mediated antiviral activity through the degradation of key factors of the virus cycle (Barr et al., 2008; Di et al., 2013; Uchil et al., 2008). TRIM22 has been reported to be involved in against of encephalomyocarditis virus (ECMV), HBV, HIV-1, and influenza A virus (Barr et al., 2008; Di et al., 2013; Hattlmann et al., 2012). In this study, we would investigate whether HCMV could regulate TRIM22 expression during HCMV primary infection. Materials and Methods Cell culture THP-1 monocytic cells were used in this study and cells were maintained in RPMI Medium 1640 (Gibco, USA) supplemented with 10% fetal calf serum (Gibco, USA), penicillin G (100 U/mL), streptomycin sulfate (100 μg/mL), l-glutamine (1%), and nonessential amino acids (1%). Cells were cultured at 37°C under 5% CO2. THP-1 monocytic cells were treated with 100 ng/mL phorbol myristate acetate (PMA) (Solarbio, Beijing, China) for 24 h to make sure that THP-1 monocytic cells had differentiated into THP-1 macrophages. Real-time PCR THP-1 macrophages were collected by using 100 μL lysis buffer (Da’an Gene Co., Ltd, China) and repeated freezing and thawing for 3 times. The mixture was boiled with 100°C for 10 min. Then the mixture was centrifuged at 13000 g for 5 min.
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影响因子：
6.7
作者：
Barr SD;Smiley JR;Bushman FD
通讯作者：
Bushman FD
DOI：
10.1038/nri2413
发表时间：
2008-11
期刊：
Nature reviews. Immunology
影响因子：
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作者：
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通讯作者：
Morse HC 3rd
DOI：
10.1155/2012/153415
发表时间：
2012
期刊：
Molecular biology international
影响因子：
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作者：
Hattlmann CJ;Kelly JN;Barr SD
通讯作者：
Barr SD
影响因子：
13
作者：
E. Vicenzi;G. Poli
通讯作者：
E. Vicenzi;G. Poli
影响因子：
5.4
作者：
M. Schwartz;Noam Stern-Ginossar
通讯作者：
M. Schwartz;Noam Stern-Ginossar