Enveloped Virus Glycoprotein/receptor Interactions

包膜病毒糖蛋白/受体相互作用

• 批准号: 8336108
• 负责人: Edward Berger
• 金额: $ 16.57万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8336108
• 关键词: B-Lymphocytes; CD209 gene; Cell Line; Cell membrane; Cells; DNA Sequence Rearrangement; Genetic; Genome; Genotype; Glycoproteins; Goals; Hepatitis C virus; Human; Human Herpesvirus 8; Infection; Infectious hepatitides; Integrins; Investigation; Length; Liver; Lytic; Mammalian Cell; Measures; Mediating; Membrane; Membrane Fusion; Methods; Molecular; Monitor; Mutation; Pathogenesis; Process; Proteins; RNA; Replicon; Reporter; Reporter Genes; Role; Signal Transduction; Slice; Staging; System; Tonsil; Tropism; Variant; Viral; Virion; Virus; Virus Diseases; Virus Receptors; Virus Replication; Virus-like particle; West Nile virus; cell type; insight; interest; novel; novel strategies; particle; prevent; receptor

We have extended our mechanistic studies of virus-receptor interactions, with focus on two specific enveloped viruses: 1) Hepatitis C virus (HCV). Using specialized mammalian producer cell line carrying a West Nile virus replicon, we devised a novel system to generate HCV virus-like particles in which all viral-encoded components (proteins and RNA) are derived from HCV. We have also used this system to produce infectious wild type HCV of virtually any genotype (confirmed for 1a, 1b, 2b). We have shown that the system can generate infectious particles carrying full-length HCV genomes that replicate in HCV-permissive target cells as well as human liver slices. Evidence suggests that the ability of the WNV-replicon cell line to secrete infectious HCV particles is due not to ongoing function of the WNV replicon, but rather to intracellular changes (presumably including membrane rearrangements) induced by the replicon. The system can also be used to generate HCV particles with specific mutations of interest, to study their effects on the HCV replication cycle. Variations of this system have been developed to measure entry of HCV VLPs wherein the introduction of a single RNA molecule from an infecting particle generates a robust reporter gene signal. The system has the advantage that the signal is dependent only on HCV entry, and not on downstream steps in the virus replication cycle; thus entry can be studied in various cell types, whether or not they can support downstream HCV replication steps. 2) Kaposi's sarcoma-associated herpesvirus (KSHV, human herpesvirus 8). we have begun to apply our studies to a recently described system wherein KSHV infection of primary B cells from human tonsils can be infected with KSHV, including reporter viruses encoding EGFP and RFP to monitor latent and lytic stages, respectively. Our goals are to analyze KSHV entry mechanisms in these primary B cells (low pH entry, direct plasma membrane fusion) as well as to determine the relevant cellular receptors, including assessment of the roles of the various previously implicated receptors (integrins, DC-SIGN, xCT).

我们已经扩展了对病毒受体相互作用的机理研究，重点是两个特定的包膜病毒：1）乙型肝炎病毒（HCV）。使用带有西尼罗河病毒复制子的专门哺乳动物生产者细胞系，我们设计了一个新型系统来生成HCV病毒样颗粒，其中所有病毒编码成分（蛋白质和RNA）均来自HCV。我们还使用该系统来生产几乎任何基因型的传染性野生型HCV（确认为1A，1B，2B）。我们已经表明，该系统可以产生携带全长HCV基因组的传染性颗粒，这些颗粒在HCV抗药性靶细胞和人肝切片中复制。有证据表明，WNV替代细胞系分泌感染性HCV颗粒的能力是由于复制子引起的WNV复制子的持续功能，而是由于wnv复制子的持续功能（大概是包括膜重排）。该系统还可以用于生成具有特定感兴趣突变的HCV颗粒，以研究其对HCV复制周期的影响。已经开发了该系统的变化来测量HCV VLP的进入，其中从感染粒子中引入单个RNA分子会产生强大的报告基因信号。该系统的优势是信号仅取决于HCV的进入，而不取决于病毒复制周期的下游步骤。因此，可以在各种单元格中研究进入，无论它们是否可以支持下游HCV复制步骤。 2）Kaposi的肉瘤相关疱疹病毒（KSHV，人类疱疹病毒8）。我们已经开始将研究应用于最近描述的系统，其中可以将来自人类扁桃体的原代B细胞感染受到KSHV感染，包括编码EGFP和RFP的报告病毒，分别监测潜在和裂解阶段。我们的目标是分析这些原代B细胞中的KSHV进入机制（低pH进入，直接质膜融合），并确定相关的细胞受体，包括评估各种先前涉及的受体（整合素，DC-SIGN，XCT）的作用。