Modulation of B-cell exosome proteins by gamma herpesvirus infection
Modulation of B-cell exosome proteins by gamma herpesvirus infection
复制标题
DOI：
10.1073/pnas.1303906110
复制
发表时间：
2013-07-30
影响因子：
11.1
通讯作者：
Raab-Traub, Nancy
中科院分区：
文献类型：
--
作者：
Meckes, David G., Jr.;Gunawardena, Harsha P.;Raab-Traub, Nancy
人类γ疱疹病毒、卡波西肉瘤相关病毒（KSHV）和EBV与多种癌症相关。最近的证据表明，EBV和可能的其他病毒可以通过将特定的病毒和细胞组分分泌到外泌体中来操纵肿瘤微环境，外泌体是从细胞释放的小的内吞衍生的囊泡。由EBV感染的鼻咽癌细胞产生的外泌体含有高水平的病毒致癌基因潜伏膜蛋白1和激活受体细胞中关键信号通路的病毒microRNA。在这项研究中，为了确定EBV和KSHV对外泌体含量的影响，对从未感染、感染EBV或KSHV或感染两种病毒的11个B细胞系纯化的外泌体进行定量蛋白质组学技术。使用质谱法，鉴定了871种蛋白质，其中类似于360种是病毒外泌体所特有的。通过2D差异凝胶电泳和光谱计数分析，与未感染的对照细胞相比以及病毒组之间鉴定出多个显著变化。这些数据预测EBV和KSHV外泌体可能调节细胞死亡和存活、核糖体功能、蛋白质合成和哺乳动物雷帕霉素靶信号传导。对外来体的不同病毒特异性作用表明，KSHV外来体将影响细胞代谢，而EBV外来体将激活通过整联蛋白、肌动蛋白、IFN和NF κ B介导的细胞信号传导。本研究中鉴定的外泌体含量的变化表明这些致癌病毒调节肿瘤微环境的方式，并可能提供EBV和KSHV相关恶性肿瘤的特异性诊断标志物。
The human gamma herpesviruses, Kaposi sarcoma-associated virus (KSHV) and EBV, are associated with multiple cancers. Recent evidence suggests that EBV and possibly other viruses can manipulate the tumor microenvironment through the secretion of specific viral and cellular components into exosomes, small endocytically derived vesicles that are released from cells. Exosomes produced by EBV-infected nasopharyngeal carcinoma cells contain high levels of the viral oncogene latent membrane protein 1 and viral microRNAs that activate critical signaling pathways in recipient cells. In this study, to determine the effects of EBV and KSHV on exosome content, quantitative proteomics techniques were performed on exosomes purified from 11 B-cell lines that are uninfected, infected with EBV or with KSHV, or infected with both viruses. Using mass spectrometry, 871 proteins were identified, of which similar to 360 were unique to the viral exosomes. Analysis by 2D difference gel electrophoresis and spectral counting identified multiple significant changes compared with the uninfected control cells and between viral groups. These data predict that both EBV and KSHV exosomes likely modulate cell death and survival, ribosome function, protein synthesis, and mammalian target of rapamycin signaling. Distinct viral-specific effects on exosomes suggest that KSHV exosomes would affect cellular metabolism, whereas EBV exosomes would activate cellular signaling mediated through integrins, actin, IFN, and NF kappa B. The changes in exosome content identified in this study suggest ways that these oncogenic viruses modulate the tumor microenvironment and may provide diagnostic markers specific for EBV and KSHV associated malignancies.