INITIATION OF CELL INFECTION BY EPSTEIN-BARR VIRUS

Epstein-Barr 病毒引发细胞感染

• 批准号: 6349775
• 负责人: Lindsey M. Hutt-Fletcher
• 金额: $ 26.16万
• 依托单位: UNIVERSITY OF MISSOURI KANSAS CITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-01-01 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6349775
• 关键词: B lymphocyte; Epstein Barr virus; epithelium; human tissue; laboratory mouse; laboratory rabbit; membrane proteins; tissue /cell culture; transfection; virus envelope; virus infection mechanism; virus protein; virus receptors; virus replication

Epstein-Barr virus (EBV) is best known for its ability to infect and cause disease by immortalizing B lymphocytes. However, the virus also accesses epithelial cells with devastating consequences. It is strongly implicated in the development of nasopharyngeal carcinoma and may play a role in the emergence of certain gastric cancers. The long term goal of this research is to understand the molecular basis for the tissue tropism of EBV. Most is known about how EBV enters B cells. A current model proposes that virus uses minimally an attachment protein gp350/220 which binds to the complement receptor type 2 (CR2) and a complex of three glycoproteins gH-gL-gp42 which is involved in penetration. Successful penetration requires that gp42 bind to HLA class II which functions as a coreceptor on the B cell surface. Entry into epithelial cells is very different. A current model for epithelial cells proposes that entry requires neither use of CR2 nor an interaction between gp42 and HLA class II. Instead, new findings suggest the existence of both a novel receptor and a novel coreceptor on epithelial cells. The immediate goal of this application is to test these models with four specific aims. The first aim is to identify the coreceptor that enables a virus that lacks gp42 to infect epithelial cells. The second aim is to explore and compare the interactions of the gH complex with B cells and epithelial cells. The third aim is to identify the viral glycoprotein used by EBV to attach to CR2- negative epithelial cells. The final aim is to identify the receptor used by EBV to infect CR2-negative epithelial cells. Identification of receptor and coreceptor will be addressed by transfecting appropriate cells with epithelial cell cDNA expression libraries. Transfected cells will be probed with recombinant viruses lacking gp42, or expressing green fluoroscent protein. Interactions of cells with the gH complex will be explored by mutational analysis and complementation of virus lacking gH. Recombinant proteins, antibodies and viruses lacking a variety of glycoproteins will be used to identify the epithelial cell attachment protein. Dissection of the molecular events that enable EBV to access the cells it infects is critical to understanding the biology of this important human pathogen.

爱泼斯坦-巴尔病毒(EBV)以其通过使B淋巴细胞永生而感染和致病的能力而闻名。然而，该病毒也会侵入上皮细胞，造成毁灭性的后果。它与鼻咽癌的发展密切相关，并可能在某些胃癌的发生中发挥作用。这项研究的长期目标是了解EBV组织趋向性的分子基础。大多数人都知道EBV是如何进入B细胞的。目前的一个模型认为，病毒最少使用与补体受体2型(CR2)结合的附着蛋白gp350/220和参与穿透的三种糖蛋白复合体Gh-g1-GP42。成功的穿透需要GP42与作为B细胞表面辅助受体的人类白细胞抗原II类结合。进入上皮细胞是非常不同的。目前的一种上皮细胞模型认为，进入细胞既不需要使用CR2，也不需要GP42和人类白细胞抗原II类分子之间的相互作用。相反，新的发现表明，上皮细胞上既存在新的受体，也存在新的辅助受体。这个应用程序的直接目标是用四个特定目标测试这些模型。第一个目标是确定使缺乏GP42的病毒感染上皮细胞的辅助受体。第二个目的是探索和比较GH复合体与B细胞和上皮细胞的相互作用。第三个目的是鉴定EB病毒用来附着在CR2阴性上皮细胞上的病毒糖蛋白。最终目的是确定EBV用来感染CR2阴性上皮细胞的受体。受体和辅助受体的鉴定将通过用上皮细胞表达文库导入适当的细胞来解决。将用缺乏GP42的重组病毒或表达绿色荧光蛋白的重组病毒来探测转基因细胞。细胞与Gh复合体的相互作用将通过突变分析和缺乏Gh的病毒的互补来探索。缺乏多种糖蛋白的重组蛋白、抗体和病毒将被用来鉴定上皮细胞附着蛋白。剖析使EBV能够访问其感染的细胞的分子事件对于了解这种重要的人类病原体的生物学至关重要。