Phenotypic analysis of latently infected CD4+ T cells.

潜伏感染的 CD4 T 细胞的表型分析。

• 批准号: 8713921
• 负责人: JOEL N BLANKSON
• 金额: $ 20.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-05 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8713921
• 关键词: Antibodies; BCL2 gene; Binding; CD4 Positive T Lymphocytes; Cell Surface Proteins; Cell surface; Cells; DNA; Development; Dose; Frequencies; Gene Expression; HIV; HIV Infections; HIV-1; Highly Active Antiretroviral Therapy; In Vitro; Individual; Infection; Lead; Libraries; Life; Mitogens; Modeling; Monoclonal Antibodies; Patients; Phase; Phenotype; Physiology; Population; Regimen; Rest; Sorting - Cell Movement; Surface; System; T-Lymphocyte; Toxin; Tumor Debulking; Viral Genes; Virus; Vorinostat; Work; antibody conjugate; antiretroviral therapy; cellular transduction; chemotherapy; gag Gene Products; in vivo; memory CD4 T lymphocyte; novel; optimism; physiologic model; public health relevance; screening; small molecule; tumor

DESCRIPTION: There has been a lot of optimism about potentially eradicating HIV-1 using strategies such as reactivation of latently infected cells by small molecules. A recent study has suggested that this approach may in fact be possible; a single dose of the molecule vorinostat was shown to induce HIV expression in CD4+ T cells in vivo. While this strategy may end up being effective, it may be very challenging to reactivate every last latently infected CD4+ T cell. A complementary strategy may be to physically eliminate latently infected cells using antibodies conjugated to toxins. This may be analogous to surgically "debulking" a tumor prior to initiating chemotherapy. However, targeting latently infected CD4+ T cells has been a challenging task because the phenotype of these cells remains largely unknown. In this new proposal, we plan to use antibodies to more than 200 different T cell markers in order to determine the phenotype of latently infected CD4+ T cells. Such an approach has been successfully used to determine the phenotype of activated T cells in a recent study. This will be analogous to screening for small molecules that activate latently infected CD4+ T cells, but in this case, a library of monoclonal antibodies (Mabs) rather than compounds will be used and the readout will be binding of Mabs to infected resting CD4+ T cells or recently reactivated latently infected CD4+ T cells rather than reactivation itself. We plan to validate our results by sorting resting CD4+ T cells from HIV-1 infected patients on suppressive HAART regimens. We specifically will sort for markers that are identified in our screen to determine whether there is an overrepresentation of HIV-1 DNA and latent replication-competent virus in these cells. This work will have major implications for strategies for HIV-1 eradication since it will potentially lead to the clearance of latently infectd cells.

描述：对于使用小分子重新激活潜伏感染细胞等策略有可能根除HIV-1，人们有很多乐观的看法。最近的一项研究表明，这种方法实际上是可能的；单剂量的涡旋剂分子被证明在体内诱导CD4+T细胞中HIV的表达。虽然这一策略最终可能是有效的，但重新激活每一个潜伏感染的CD4+T细胞可能是非常具有挑战性的。 一种补充策略可能是使用与毒素结合的抗体来物理消除潜伏感染的细胞。这可能类似于在开始化疗前对肿瘤进行手术“去瘤”。然而，靶向潜伏感染的CD4+T细胞一直是一项具有挑战性的任务，因为这些细胞的表型在很大程度上仍不清楚。在这项新提议中，我们计划使用200多种不同T细胞标志物的抗体来确定潜伏感染的CD4+T细胞的表型。在最近的一项研究中，这种方法已经成功地用于确定激活的T细胞的表型。这将类似于筛选激活潜伏感染的CD4+T细胞的小分子，但在这种情况下，将使用单抗(MAb)库而不是化合物，读出的将是单抗与感染的静止的CD4+T细胞或最近重新激活的潜伏感染的CD4+T细胞的结合，而不是 重新激活本身。我们计划通过在抑制性HAART方案中对HIV-1感染患者的静止CD4+T细胞进行分选来验证我们的结果。我们特别将筛选在我们的屏幕上识别的标记，以确定这些细胞中是否存在HIV-1DNA和潜伏复制能力病毒的过度表达。这项工作将对根除艾滋病毒-1的战略产生重大影响，因为它可能导致清除潜伏感染的细胞。