IDENTIFICATION OF CD8+ CELL ANTI HIV FACTOR

CD8细胞抗HIV因子的鉴定

• 批准号: 7369044
• 负责人: JAY A LEVY
• 金额: $ 1.89万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7369044
• 关键词: IDENTIFICATION; CD8+; CELL; ANTI; HIV

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Introduction Individuals who have been infected with HIV and remain healthy for over 10 years are considered long-term survivors. These asymptomatic individuals have a strong cell-mediated immune response that suppresses HIV replication. This activity is lost as the infected person advances to disease. The antiviral response is mediated by a secreted CD8+ cell antiviral factor (CAF) which blocks transcription via the HIV promoter. CAF is produced at low levels by CD8+ cells from these healthy infected individuals. The protein appears to be unlike any of the known cytokines, chemokines and human growth factors. CAF identification will most likely be achieved through protein purification and mass spectrometry. Methods CD8+ cells from healthy HIV-infected individuals are grown in culture medium lacking serum and where possible, albumin. Fluids are assayed for antiviral activity by standard procedures in the laboratory. Active fluids are then concentrated and fractionated by various biochemical techniques including ion exchange, chromatography, and by sizing columns Fractions with enriched CAF activity are analyzed by 2-D gel and evaluated by mass spectrometry. Results Thus far, we have been able to show that CAF has a molecular size of about 50 kD. By differential 2D gel analysis, there appears to be at least 20 different spots that are associated with CAF activity. Studies are in progress to analyzed by mass spectrometry. Those protein spots that are unique to CAF-containing fluids will be further evaluated. Discussion By mass spectrometry, we hope to be able to identify the protein (CAF) that mediates this anti-HIV activity.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构适用于该中心，这不一定是调查员的机构。引言已经感染了艾滋病毒并保持健康已有10多年的人被认为是长期幸存者。这些无症状的个体具有抑制HIV复制的强细胞介导的免疫反应。随着感染者发展为疾病，这项活动失去了。抗病毒反应是由分泌的CD8+细胞抗病毒因子（CAF）介导的，该因子通过HIV启动子阻止转录。 CAF是由这些健康感染个体的CD8+细胞低水平产生的。该蛋白质似乎与任何已知的细胞因子，趋化因子和人类生长因子不同。 CAF鉴定很可能是通过蛋白质纯化和质谱法实现的。 来自健康HIV感染个体的方法CD8+细胞在缺乏血清的培养基中生长，并且在可能的情况下，白蛋白。通过实验室中的标准程序测定液体的抗病毒活性。然后，通过各种生化技术将活性流体浓缩，并通过包括离子交换，色谱法以及通过富含Caf活性的尺寸柱分数进行分馏，并通过2-D凝胶分析，并通过质谱法评估。 结果到目前为止，我们已经能够证明CAF的分子大小约为50 kD。通过差异2D凝胶分析，似乎至少有20个与CAF活性相关的不同斑点。正在进行研究以通过质谱分析。将进一步评估那些含CAF流体的蛋白质斑点。 通过质谱法讨论，我们希望能够鉴定介导这种抗HIV活性的蛋白质（CAF）。