Department of Molecular and Cell Biology, University of Connecticut, 91 North Eagleville Road, BSP 318, Storrs, CT 06269, USA.
Biosens Bioelectron. 2012 Jan 15;31(1):264-9. doi: 10.1016/j.bios.2011.10.029. Epub 2011 Oct 29.
The circulating population of peripheral T lymphocytes obtained from a blood sample can provide a large amount of information about an individual's medical status and history. Recent evidence indicates that the detection and functional characterization of antigen-specific T cell subsets within the circulating population may provide a diagnostic indicator of disease and has the potential to predict an individual's response to therapy. In this report, a microarray detection platform that combines grating-coupled surface plasmon resonance imaging (GCSPRI) and grating-coupled surface plasmon coupled emission (SPCE) fluorescence detection modalities were used to detect and characterize CD4(+) T cells. The microspot regions of interest (ROIs) printed on the array consisted of immobilized antibodies or peptide loaded MHC monomers (p/MHC) as T cell capture ligands mixed with additional antibodies as cytokine capture ligands covalently bound to the surface of a corrugated gold sensor chip. Using optimized parameters, an unlabeled influenza peptide reactive T cell clone could be detected at a frequency of 0.1% in a mixed T cell sample using GCSPRI. Additionally, after cell binding was quantified, differential TH1 cytokine secretion patterns from a T cell clone cultured under TH1 or TH2 inducing conditions was detected using an SPCE fluorescence based assay. Differences in the secretion patterns of 3 cytokines, characteristic of the inducing conditions, indicated that differences were a consequence of the functional status of the captured cells. A dual mode GCSPRI/SPCE assay can provide a rapid, high content T cell screening/characterization tool that is useful for diagnosing disease, evaluating vaccination efficacy, or assessing responses to immunotherapeutics.
从血液样本中获得的外周循环 T 淋巴细胞可以提供大量有关个体医疗状况和病史的信息。最近的证据表明，在循环人群中检测和功能表征抗原特异性 T 细胞亚群可能提供疾病的诊断指标，并有可能预测个体对治疗的反应。在本报告中，使用结合光栅耦合表面等离子体共振成像（GCSPRI）和光栅耦合表面等离子体耦合发射（SPCE）荧光检测模式的微阵列检测平台来检测和表征 CD4(+) T 细胞。阵列上的微点感兴趣区域（ROI）由固定化抗体或负载肽的 MHC 单体（p/MHC）作为 T 细胞捕获配体组成，与作为细胞因子捕获配体的额外抗体混合，共价结合到波纹金传感器芯片的表面。使用优化的参数，在混合 T 细胞样品中，可以使用 GCSPRI 以 0.1%的频率检测未标记的流感肽反应性 T 细胞克隆。此外，在量化细胞结合后，使用基于 SPCE 荧光的测定法检测在 TH1 或 TH2 诱导条件下培养的 T 细胞克隆的差异 TH1 细胞因子分泌模式。诱导条件特征的 3 种细胞因子分泌模式的差异表明，差异是由于捕获细胞的功能状态所致。双模式 GCSPRI/SPCE 测定法可提供快速、高内涵的 T 细胞筛选/表征工具，可用于诊断疾病、评估疫苗效力或评估免疫治疗的反应。
