Virus-Specific Nanobody-Chimeras Degrade the Human Cytomegalovirus US28 Protein in CD34+ Cells.

IF 3.3 3区 医学 Q2 MICROBIOLOGY
Emma Poole, Janika Schmitt, Stephen C Graham, Bernard T Kelly, John Sinclair
{"title":"Virus-Specific Nanobody-Chimeras Degrade the Human Cytomegalovirus US28 Protein in CD34+ Cells.","authors":"Emma Poole, Janika Schmitt, Stephen C Graham, Bernard T Kelly, John Sinclair","doi":"10.3390/pathogens13100821","DOIUrl":null,"url":null,"abstract":"<p><p>After primary infection, human cytomegalovirus (HCMV) establishes lifelong persistence, underpinned by latent carriage of the virus with spontaneous reactivation events. In the immune-competent, primary infection or reactivation from latency rarely causes disease. However, HCMV can cause significant disease in immune-compromised individuals such as immune-suppressed transplant patients. Latency, where the viral genome is carried in the absence of the production of infectious virions, can be established in undifferentiated cells of the myeloid lineage. A number of stimuli can cause virus reactivation from latency to occur, beginning with the induction of viral immediate-early (IE) lytic gene expression. The suppression of viral IE gene expression to establish and maintain latent infection is known to result from a balance of viral and cellular factors. One key viral factor involved in this is the G protein-coupled receptor US28. Recently, we have shown that US28 is targeted for degradation by a modified nanobody (PCTD-Vun100bv) based on the novel PACTAC (PCSK9-antibody clearance-targeting chimeras) approach for targeted protein degradation. Furthermore, we have shown that this PCTD-Vun100bv-induced degradation of US28 results in IE gene expression in experimentally latently infected CD14+ monocytes. However, HCMV also establishes latency in CD34+ bone marrow cells, the progenitors of CD14+ cells. Here, we show that PCTD-Vun100bv also causes US28 degradation in these CD34+ primary cells, again resulting in the induction of viral IE gene expression. Additionally, we show that PCTD-Vun100bv can target US28 in naturally latently infected CD14+ monocytes from an HCMV-seropositive donor, allowing these latently infected cells to be killed by HCMV-specific cytotoxic T cells from that same donor. These observations support the view that targeting US28 for degradation during natural latency could be a tractable 'shock-and-kill' strategy to target the latent HCMV reservoir in myeloid cells.</p>","PeriodicalId":19758,"journal":{"name":"Pathogens","volume":null,"pages":null},"PeriodicalIF":3.3000,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11510245/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Pathogens","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3390/pathogens13100821","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

After primary infection, human cytomegalovirus (HCMV) establishes lifelong persistence, underpinned by latent carriage of the virus with spontaneous reactivation events. In the immune-competent, primary infection or reactivation from latency rarely causes disease. However, HCMV can cause significant disease in immune-compromised individuals such as immune-suppressed transplant patients. Latency, where the viral genome is carried in the absence of the production of infectious virions, can be established in undifferentiated cells of the myeloid lineage. A number of stimuli can cause virus reactivation from latency to occur, beginning with the induction of viral immediate-early (IE) lytic gene expression. The suppression of viral IE gene expression to establish and maintain latent infection is known to result from a balance of viral and cellular factors. One key viral factor involved in this is the G protein-coupled receptor US28. Recently, we have shown that US28 is targeted for degradation by a modified nanobody (PCTD-Vun100bv) based on the novel PACTAC (PCSK9-antibody clearance-targeting chimeras) approach for targeted protein degradation. Furthermore, we have shown that this PCTD-Vun100bv-induced degradation of US28 results in IE gene expression in experimentally latently infected CD14+ monocytes. However, HCMV also establishes latency in CD34+ bone marrow cells, the progenitors of CD14+ cells. Here, we show that PCTD-Vun100bv also causes US28 degradation in these CD34+ primary cells, again resulting in the induction of viral IE gene expression. Additionally, we show that PCTD-Vun100bv can target US28 in naturally latently infected CD14+ monocytes from an HCMV-seropositive donor, allowing these latently infected cells to be killed by HCMV-specific cytotoxic T cells from that same donor. These observations support the view that targeting US28 for degradation during natural latency could be a tractable 'shock-and-kill' strategy to target the latent HCMV reservoir in myeloid cells.

病毒特异性纳米抗体-嵌合体能降解 CD34+ 细胞中的人类巨细胞病毒 US28 蛋白。
人类巨细胞病毒(HCMV)在原发感染后会终生潜伏,潜伏期内病毒会自发重新激活。在免疫功能正常的人群中,原发感染或潜伏期再活化很少会导致疾病。然而,HCMV 可在免疫力低下的人群(如免疫抑制移植患者)中引起严重疾病。潜伏期是指病毒基因组在不产生感染性病毒的情况下携带病毒,可在未分化的髓系细胞中建立。一些刺激因素可导致病毒从潜伏期重新活化,首先是诱导病毒即刻早期(IE)溶解基因的表达。抑制病毒 IE 基因表达以建立和维持潜伏感染,是病毒和细胞因素平衡的结果。其中一个关键的病毒因子是 G 蛋白偶联受体 US28。最近,我们发现 US28 可被一种基于新型 PACTAC(PCSK9-抗体清除-靶向嵌合体)方法的改良纳米抗体(PCTD-Vun100bv)靶向降解。此外,我们还证明,PCTD-Vun100bv 诱导的 US28 降解会导致潜伏感染的 CD14+ 单核细胞中 IE 基因的表达。然而,HCMV 也会在 CD34+ 骨髓细胞(CD14+ 细胞的祖细胞)中潜伏。在这里,我们发现 PCTD-Vun100bv 也会导致这些 CD34+ 原始细胞中的 US28 降解,从而再次诱导病毒 IE 基因的表达。此外,我们还发现 PCTD-Vun100bv 可以靶向 HCMV 血清阳性供体中自然潜伏感染的 CD14+ 单核细胞中的 US28,从而使这些潜伏感染的细胞被来自同一供体的 HCMV 特异性细胞毒性 T 细胞杀死。这些观察结果支持这样一种观点,即在自然潜伏期靶向降解 US28 可能是针对髓系细胞中潜伏 HCMV 库的一种可行的 "冲击-杀伤 "策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Pathogens
Pathogens Medicine-Immunology and Allergy
CiteScore
6.40
自引率
8.10%
发文量
1285
审稿时长
17.75 days
期刊介绍: Pathogens (ISSN 2076-0817) publishes reviews, regular research papers and short notes on all aspects of pathogens and pathogen-host interactions. There is no restriction on the length of the papers. Our aim is to encourage scientists to publish their experimental and theoretical research in as much detail as possible. Full experimental and/or methodical details must be provided for research articles.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信