GENETICS AND FUNCTIONS OF KSHV GLYCOPROTEINS GM AND GN

KSHV 糖蛋白 GM 和 GN 的遗传学和功能

• 批准号: 7959460
• 负责人: OSWALD D'AUVERGNE
• 金额: $ 9.61万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959460
• 关键词: B-Cell Lymphomas; Biomedical Research; Body cavities; Cell Line; Cells; Codon Nucleotides; Computer Retrieval of Information on Scientific Projects Database; DEAE-Dextran; Funding; Genes; Glycoproteins; Grant; Human Herpesvirus 8; Infection; Institution; Kaposi Sarcoma; Louisiana; Lymphoma; Lysine; Measurement; Multicentric Angiofollicular Lymphoid Hyperplasia; Mus; Nude Mice; Oncogenic Viruses; Pathogenesis; Pleural effusion disorder; Polybrene; Protamine Sulfate; Protamines; Protein Biosynthesis; Research; Research Personnel; Resources; Role; Small Interfering RNA; Source; System; Time; Transfection; United States National Institutes of Health; Viral; Virion; Virus; Work; base; body cavity; effusion; graduate student; matrigel; polycation; reconstitution; tumor; tumorigenesis

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Kaposi's sarcoma-associated herpesvirus (KSHV), or human herpesvirus 8, is a lymphotropic oncogenic virus that has been implicated in the pathogenesis of Kaposi's sarcoma; body cavity-based B-cell lymphoma (BCBL), or primary effusion lymphoma; and some forms of multicentric Castleman's disease. We have developed a conditional silencing system for partial inhibition of KSHV viral glycoprotein synthesis using specific siRNAs. In this system, protein synthesis can be reconstituted using codon-optimized genes that are not susceptible to siRNA inhibition. We have succesfully used this system to demonstrate that KSHV glycoprotein B (gB) is necessary for virion egress from BCBL-1 cells and virus infectivity (Subramanian, D'Auvergne, et al. J. Virol. 2008, 82:7144-54). To investigate the potential role of gB in tumorigenesis, BCBL-1 cells transiently transfected with anti-gB siRNAs and codon optimized gB were mixed with matrigel and injected subcutaneously in nude mice. Direct measurement of tumors revealed that BCBL-1 cells transfected with anti-gB siRNAs produced tumors significantly smaller than mock-transfected BCBL-1 cells. Co-transfection of codon optimized gB appeared to abrogate the inhibition of tumor formation by siRNAs. These results show that gB is important for infectivity, virion egress and pleural effusion lymphoma (PEL) formation in mice. As part of a graduate student project, we have also worked on the effect of polycations on facilitating KSHV virus entry into 293 cells. It was found that DEAE-dextran, poly-lysine, polybrene, protamine and protamine sulfate enhanced the KSHV entry into 293 and other cell lines when added at the time of infection.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 卡波西肉瘤相关疱疹病毒（Kaposi's sarcoma-associated herpesvirus，KSHV），又称人类疱疹病毒8型（human herpesvirus 8），是一种嗜淋巴细胞的致癌病毒，与卡波西肉瘤、体腔B细胞淋巴瘤（body cavity-based B-cell lymphoma，BCBL）或原发性渗出性淋巴瘤（primary effusion lymphoma）以及某些形式的多中心Castleman病（multicenter Castleman's disease）的发病机制有关。我们开发了一种条件性沉默系统，使用特异性SiRNA部分抑制KSHV病毒糖蛋白的合成。在该系统中，可以使用对siRNA抑制不敏感的密码子优化的基因来重建蛋白质合成。我们已经成功地使用该系统来证明KSHV糖蛋白B（gB）对于病毒体从BCBL-1细胞中排出和病毒感染性是必需的（Subramanian，D ′ Auvergne，et al.J.Virol. 2008，82：7144-54）。为了研究gB在肿瘤发生中的潜在作用，将用抗gB siRNA和密码子优化的gB瞬时转染的BCBL-1细胞与基质胶混合并皮下注射到裸鼠中。肿瘤的直接测量显示，用抗gB siRNA转染的BCBL-1细胞产生的肿瘤显著小于模拟转染的BCBL-1细胞。密码子优化的gB的共转染似乎消除了siRNA对肿瘤形成的抑制。这些结果表明，gB是重要的感染性，病毒粒子出口和胸腔积液淋巴瘤（PEL）的形成在小鼠中。作为研究生项目的一部分，我们还研究了聚阳离子对促进KSHV病毒进入293细胞的影响。结果发现，当在感染时加入DEAE-葡聚糖、聚赖氨酸、聚凝胺、鱼精蛋白和硫酸鱼精蛋白时，KSHV进入293和其它细胞系中增强。