ROLE OF GLYCOSPHINGOLIPIDS IN HIV 1 INFECTION OF THE CNS

鞘糖脂在 HIV 1 中枢神经系统感染中的作用

• 批准号: 6313406
• 负责人: RICHARD D KENSINGER
• 金额: $ 1.96万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6313406
• 关键词: AIDS dementia complex; HIV envelope protein gp120; antiAIDS agent; carbohydrates; cell line; ceramides; drug design /synthesis /production; enzyme linked immunosorbent assay; glycosphingolipids; human immunodeficiency virus 1; intermolecular interaction; recombinant proteins; thin layer chromatography; virus receptors

The mechanism by which HIV-associated dementia develops is unclear. It is known, however, that astrocytes, neurons, and brain capillary endothelial cells can be infected in a CD4- independent pathway, in which glycosphingolipids (GSLs) are thought to serve as receptors for HIV. Although CD4 expressing macrophages and microglial cells are the main reservoirs for HIV in the CNS, GSLs were shown to be needed for gp120 mediated fusion of CD4 expressing cells. The overall goal of the proposed research is to determine the role of GSLs in the attachment/fusion process of HIV-1 infection of CNS cells, and to inhibit that interaction. The specific aims of this proposal are to: 1) Confirm that GSLs are necessary for HIV-1 infection, and identify which GSLs are adhered to best by recombinant soluble gp120 and purified virus. This will be accomplished by determining the ability of purified GSLs, isolated from neural cells known to be infected by HIV-1, to function as ligands using TLC overlay and ELISA assays. 2) Determine which portion of the GSL(s) identified in Aim 1, interacts with gp120-the oligosaccharide, ceramide, or both. The oligosaccharide portion of the best GSL ligand will be tested for its ability to inhibit the adherence of gp120 to the immobilized GSL ligand. 3) Prepare a multivalent binding inhibitor based on the GSL moiety found to interact with gpl20 in Aim 2. Then, determine the ability of the synthesized inhibitor to block gp120 adherence to the immobilized GSL ligand, and HIV-1 entry into cultured neural cells.

尚不清楚与HIV相关的痴呆发育的机制尚不清楚。 然而，众所周知，星形胶质细胞，神经元和脑毛细血管内皮细胞可以在CD4独立途径中感染，其中糖磷脂脂（GSL）被认为是HIV的受体。 尽管表达巨噬细胞和小胶质细胞的CD4是CNS中HIV的主要储藏，但GSL被证明是GP120介导的CD4表达细胞融合所需的。 拟议的研究的总体目标是确定GSL在CNS细胞HIV-1感染的附着/融合过程中的作用，并抑制这种相互作用。 该提案的具体目的是：1）确认GSL对于HIV-1感染是必需的，并确定通过重组可溶性GP120和纯化病毒遵守哪些GSL。 这将通过确定从已知被HIV-1感染的神经细胞中分离出的纯化GSL的能力来实现，该能力使用TLC叠加层和ELISA测定法功能充当配体。 2）确定在AIM 1中鉴定出的GSL的哪一部分，与GP120-寡糖，神经酰胺或两者相互作用。 最佳GSL配体的寡糖部分将通过抑制GP120对固定的GSL配体的粘附能力进行测试。 3）根据发现在AIM 2中与GPL20相互作用的GSL部分的多价结合抑制剂准备。