Optical Chip Test for T Cell Function in HIV Patients

描述(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，该平台基于第一阶段计划产生的芯片上实时扩增检测(OCRAD)方法。目前进行基因表达分析的方法需要冗长而复杂的程序，包括单独的RNA扩增、杂交、洗涤和标记读出步骤。OCRAD采用折射式集成光学芯片(IOC)传感器进行多路芯片上核酸放大。这种芯片内扩增和实时检测的结合将创建一个具有必要功能的平台，使研究人员以更快、更简单和更便宜的形式进行基因表达测量的能力取得重大进步。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。CD8T细胞对许多病毒感染的控制至关重要，也是艾滋病毒疫苗努力的目标。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CD8T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很好地确定。使用基于先前使用病毒感染模型进行的基因表达研究的小基因集图谱，OCRAD将被用于分析来自HIV健康患者、HIV+病毒载量抑制患者和HIV+病毒血症患者CD8 T细胞亚群的基因表达。人们认为，这一分析可以更好地了解HIV感染过程中CD8T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，并可能发现可用作治疗干预目标的重要生物学途径。

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.