IDENTIFICATION OF CD8+ CELL ANTI HIV FACTOR

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 感染艾滋病毒并保持健康超过10年的个人被认为是长期幸存者。这些没有症状的人有一种强大的细胞介导的免疫反应，可以抑制艾滋病毒的复制。随着感染者病情的发展，这种活性会丧失。抗病毒反应是由分泌的CD8+细胞抗病毒因子(CAF)介导的，它通过HIV启动子阻止转录。CAF是由这些健康感染者的CD8+细胞低水平产生的。这种蛋白质似乎不同于任何已知的细胞因子、趋化因子和人类生长因子。CAF的鉴定最有可能通过蛋白质纯化和质谱分析来实现。 来自健康的HIV感染者的CD8+细胞在没有血清和白蛋白的培养液中培养。液体通过实验室中的标准程序进行抗病毒活性检测。然后，活性流体通过各种生化技术进行浓缩和分级，包括离子交换、层析和通过调整色谱柱大小，对具有丰富CaF活性的馏分进行二维凝胶分析和质谱分析。

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. 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. 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.