GENETICS AND FUNCTIONS OF KSHV GLYCOPROTEINS GM AND GN

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 卡波西肉瘤相关疱疹病毒(KSHV)，或人类疱疹病毒8型，是一种嗜淋巴致癌病毒，与卡波西肉瘤、体腔B细胞淋巴瘤(BCBL)或原发渗出性淋巴瘤以及某些形式的多中心Castleman病的发病有关。我们开发了一种条件沉默系统，利用特定的siRNA部分抑制KSHV病毒糖蛋白的合成。在这个系统中，蛋白质合成可以使用密码子优化的基因重组，这些基因不受siRNA抑制的影响。我们已经成功地用这个系统证明了KSHV糖蛋白B(GB)是病毒粒子从BCBL-1细胞中排出和病毒感染性所必需的(Subramanian，D‘Auvergne等人)。J.维罗尔。2008年，82：7144-54)。为探讨GB在肿瘤发生中的作用，将瞬时转染抗GB siRNAs和密码子优化的GB的BCBL-1细胞与Matrigel混合后注射到裸鼠皮下。肿瘤的直接测量显示，转染抗GB siRNAs的BCBL-1细胞产生的肿瘤明显小于模型转染的BCBL-1细胞。共转染密码子优化的GB似乎取消了siRNAs对肿瘤形成的抑制作用。这些结果表明，GB对小鼠的传染性、病毒粒子外泄和胸腔积液淋巴瘤(PEL)的形成具有重要作用。作为研究生项目的一部分，我们还研究了聚阳离子在促进KSHV病毒进入293细胞方面的作用。结果发现，DEAE-葡聚糖、多赖氨酸、聚丙稀胺、鱼精蛋白和鱼精蛋白硫酸盐在感染时加入可促进KSHV进入293和其他细胞系。

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.