HIV ENVELOPE SPECIFIC DARPIN-BASED MICROBICIDE

HIV 包膜特异性 DARPIN 杀菌剂

• 批准号: 8358133
• 负责人: Melissa J Robbiani
• 金额: $ 5.78万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358133
• 关键词: Ankyrin Repeat; Binding; Biological; Blood; Blood Circulation; CD4 Positive T Lymphocytes; Cell surface; Cells; Clinical; Dendritic Cells; Exhibits; Funding; Future; Grant; HIV; Hour; Human; In Vitro; Infection; Injection of therapeutic agent; Kinetics; Ligands; Macaca; Macaca mulatta; Methodology; National Center for Research Resources; Plasma; Primates; Principal Investigator; Production; Protein Binding; Proteins; Research; Research Infrastructure; Resources; SIV; Source; T-Lymphocyte; Technology; United States National Institutes of Health; base; cell type; cost; design; in vivo; lymph nodes; microbicide; monocyte; novel; peripheral blood

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. BACKGROUND: The recently described Designed Ankyrin Repeat Protein (DARPin) technology can produce highly selective ligands to a variety of biological targets at a low production cost. METHODOLOGY/PRINCIPAL FINDINGS: To investigate the in vivo use of DARPins for future application to novel anti-HIV strategies, we identified potent CD4-specific DARPins that recognize rhesus CD4 and followed the fate of intravenously injected CD4-specific DARPin 57.2 in rhesus macaques. The human CD4-specific DARPin 57.2 bound macaque CD4(+) cells and exhibited potent inhibitory activity against SIV infection in vitro. DARPin 57.2 or the control E3_5 DARPin was injected into rhesus macaques and the fate of cell-free and cell-bound CD4-specific DARPin was evaluated. DARPin-bound CD4(+) cells were detected in the peripheral blood as early as 30 minutes after the injection, decreasing within 6 hours and being almost undetectable within 24 hours. The amount of DARPin bound was dependent on the amount of DARPin injected. CD4-specific DARPin was also detected on CD4(+) cells in the lymph nodes within 30 minutes, which persisted with similar kinetics to blood. More extensive analysis using blood revealed that DARPin 57.2 bound to all CD4(+) cell types (T cells, monocytes, dendritic cells) in vivo and in vitro with the amount of binding directly proportional to the amount of CD4 on the cell surface. Cell-free DARPins were also detected in the plasma, but were rapidly cleared from circulation. CONCLUSIONS/SIGNIFICANCE: We demonstrated that the CD4-specific DARPin can rapidly and selectively bind its target cells in vivo, warranting further studies on possible clinical use of the DARPin technology.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 背景：最近描述的设计锚蛋白重复蛋白（DARPin）技术可以以较低的生产成本生产针对多种生物靶标的高选择性配体。 方法学/主要发现：为了研究 DARPin 的体内应用以供未来新型抗 HIV 策略的应用，我们鉴定了识别恒河猴 CD4 的有效 CD4 特异性 DARPin，并跟踪了恒河猴静脉注射 CD4 特异性 DARPin 57.2 的命运。人CD4特异性DARPin 57.2结合猕猴CD4(+)细胞，并在体外表现出对SIV感染的有效抑制活性。将 DARPin 57.2 或对照 E3_5 DARPin 注射到恒河猴中，并评估无细胞和细胞结合的 CD4 特异性 DARPin 的命运。注射后30分钟即可在外周血中检测到DARPin结合的CD4(+)细胞，6小时内减少，24小时内几乎检测不到。结合的 DARPin 量取决于注射的 DARPin 量。 30 分钟内在淋巴结中的 CD4(+) 细胞上也检测到了 CD4 特异性 DARPin，其持续存在的动力学与血液相似。使用血液进行更广泛的分析表明，DARPin 57.2 在体内和体外与所有 CD4(+) 细胞类型（T 细胞、单核细胞、树突状细胞）结合，结合量与细胞表面 CD4 的量成正比。血浆中也检测到了无细胞 DARPins，但很快就从循环中清除。结论/意义：我们证明 CD4 特异性 DARPin 可以在体内快速、选择性地结合其靶细胞，值得进一步研究 DARPin 技术的可能临床应用。