Optical Chip Test for T Cell Function in HIV Patients

描述（由申请人提供）：本提案旨在开发一种新的研究平台，用于分析HIV感染导致的免疫细胞基因表达，该平台基于芯片上实时扩增检测（ocad）方法，该方法来自I期项目。目前进行基因表达分析的方法需要冗长和复杂的协议，包括单独的RNA扩增，杂交，洗涤和标签读出步骤。ocad采用折光集成光学芯片（IOC）传感器进行多路片上核酸扩增。这种芯片上扩增和实时检测的结合将创造一个具有必要功能的平台，使研究人员能够以更快速，更简单，更便宜的方式进行基因表达测量。基因表达分析是了解细胞分化状态的重要工具，可能在临床环境中作为诊断或预测工具有用。CD8 T细胞对于控制许多病毒感染至关重要，也是HIV疫苗工作的目标。然而，在HIV感染期间，CD8 T细胞的免疫控制最终失败，导致艾滋病。大量证据表明，HIV患者的CD8 T细胞经历了不适当的分化过程，导致功能受损，尽管这一过程在人类中的分子特征尚未得到很好的定义。利用基于先前病毒感染小鼠模型的基因表达研究的小基因集谱，ocad将应用于分析HIV-健康患者、病毒载量受抑制的HIV+患者和病毒血症HIV+患者的CD8 T细胞亚群中的基因表达。相信这一分析将更好地理解HIV感染过程中CD8 T细胞的分子特征。这可能最终导致改进的诊断和/或预测临床工具，并可能发现重要的生物学途径，可作为治疗干预的目标。

DESCRIPTION (provided by applicant): This proposal is targeted at the development of a novel research platform for analyzing immune cell gene expression as a result of HIV infection that is based on the on-chip real-time amplification detection (OCRAD) method that came out of the Phase I program. Current methods for performing gene expression analysis require lengthy and complex protocols, involving separate RNA amplification, hybridization, washing and label read-out steps. OCRAD employs a refractometric integrated optic chip (IOC) sensor to perform multiplex on-chip nucleic acid amplification. This combination of on-chip amplification and real-time detection will create a platform with the features necessary to make a significant advancement in the ability of researchers to perform gene expression measurements in a more rapid, simple and less expensive format. Gene expression analysis is an important tool for understanding a cell's differentiation state and may be useful in a clinical setting as a diagnostic or predictive tool. CD8 T cells are critical for the control of many viral infections and are the target of vaccine efforts for HIV. However, during HIV infection immune control by CD8 T cells eventually fails leading to AIDS. Considerable evidence suggests that CD8 T cells in HIV patients undergo an inappropriate differentiation process leading to impaired function, although the molecular signature of this process in humans is not well defined. Using a small gene set profile based on previous gene expression studies using mouse models of viral infection, OCRAD will be applied to analyzing gene expression in CD8 T cell subsets from HIV- healthy patients, HIV+ patients with suppressed viral load and HIV+ patients with viremia. It is believed that this analysis should provide a better understanding of the molecular profile of CD8 T cells during HIV infection. This may ultimately lead to improved diagnostic and/or predictive clinical tools and may uncover important biological pathways that can be used as targets for therapeutic intervention.