Optical Chip Test for T Cell Function in HIV Patients

描述（由申请人提供）：该提案旨在开发一种新型研究平台，用于分析HIV感染导致的免疫细胞基因表达，该平台基于I期项目中的芯片实时扩增检测（OCRAD）方法。目前用于进行基因表达分析的方法需要冗长且复杂的方案，涉及单独的RNA扩增、杂交、洗涤和标记读出步骤。OCRAD采用折射集成光学芯片（IOC）传感器进行多重芯片上核酸扩增。这种芯片上扩增和实时检测的结合将创建一个平台，该平台具有必要的功能，可以显著提高研究人员以更快速，简单和更便宜的形式进行基因表达测量的能力。基因表达分析是理解细胞分化状态的重要工具，并且在临床环境中作为诊断或预测工具可能是有用的。CD 8 T细胞对于控制许多病毒感染至关重要，并且是HIV疫苗的目标。然而，在HIV感染期间，CD 8 T细胞的免疫控制最终失败，导致艾滋病。相当多的证据表明，HIV患者的CD 8 T细胞经历了不适当的分化过程，导致功能受损，尽管人类中这一过程的分子特征尚未明确。使用基于先前使用病毒感染小鼠模型的基因表达研究的小基因集谱，OCRAD将用于分析来自HIV-健康患者、具有抑制病毒载量的HIV+患者和具有病毒血症的HIV+患者的CD 8 T细胞亚群中的基因表达。据信，这种分析应该提供更好地了解HIV感染过程中CD 8 T细胞的分子特征。这可能最终导致改进的诊断和/或预测性临床工具，并可能揭示可用作治疗干预靶点的重要生物学途径。

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.