原发性血管内皮细胞中高效的人类巨细胞病毒复制依赖于 SOCS3。

Efficient Human Cytomegalovirus Replication in Primary Endothelial Cells Is SOCS3 Dependent.

机构信息

Division of Hematology/Oncology, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts, USA,

Division of Infectious Diseases and Hospital Epidemiology, University Hospital Zurich, University of Zurich, Zurich, Switzerland.

出版信息

Intervirology. 2019;62(2):80-89. doi: 10.1159/000501383. Epub 2019 Jul 17.

DOI:10.1159/000501383

PMID:31315128

Abstract

BACKGROUND

In immunocompromised patients, human cytomegalovirus (HCMV) infection is a major cause of morbidity and mortality. Suppressor of cytokine signaling (SOCS) proteins are very potent negative regulators of the janus kinase/signal transducer and activator of transcription (JAK/STAT) pathways. We hypothesized that HCMV exploits SOCS1 and/or SOCS3 to its advantage.

METHODS

All experiments were carried out with primary human lung-derived microvascular endothelial cells (HMVEC). SOCS1 and SOCS3 were silenced by transfecting the cells with siRNA. HCMV was propagated and titered on human lung-derived fibroblasts MRC5. Real-time PCR and Western blot were used to detect mRNA and protein levels, respectively.

RESULTS

The data presented show that an efficient replication of HCMV in HMVEC is dependent on SOCS3 protein. Time course analysis revealed an increase in SOCS3 protein levels in infected cells. Silencing of SOCS3 (siSOCS3) resulted in inhibition of viral immediate early, early, and late antigen production. Consistently, HCMV titers produced by siSOCS3 cultures were significantly decreased when compared to control transfected cultures (siCNTRs). STAT1 and STAT2 phosphorylation was increased in siSOCS3-infected cells when compared to siCNTR-treated cells.

CONCLUSION

These findings indicate the implication of SOCS3 in the mechanism of HCMV-mediated control of cellular immune responses.

摘要

背景

在免疫功能低下的患者中，人巨细胞病毒（HCMV）感染是发病率和死亡率的主要原因。细胞因子信号转导抑制因子（SOCS）蛋白是 Janus 激酶/信号转导和转录激活因子（JAK/STAT）途径的非常有效的负调控因子。我们假设 HCMV 利用 SOCS1 和/或 SOCS3 为其所用。

方法

所有实验均使用原代人肺微血管内皮细胞（HMVEC）进行。通过转染细胞用 siRNA 沉默 SOCS1 和 SOCS3。在人肺衍生成纤维细胞 MRC5 上繁殖和滴定 HCMV。实时 PCR 和 Western blot 分别用于检测 mRNA 和蛋白质水平。

结果

呈现的数据表明，HCMV 在 HMVEC 中的有效复制依赖于 SOCS3 蛋白。时间进程分析显示感染细胞中 SOCS3 蛋白水平增加。SOCS3 沉默（siSOCS3）导致病毒即刻早期、早期和晚期抗原产生受到抑制。一致地，与对照转染培养物（siCNTRs）相比，siSOCS3 培养物产生的 HCMV 滴度显着降低。与 siCNTR 处理的细胞相比，siSOCS3 感染的细胞中 STAT1 和 STAT2 磷酸化增加。