Label-free detection of ConA-induced T-lymphocyte activation at single-cell level by microfluidics.

IF 3 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Yameng Liu, Xiaohu Wang, Yuxia Lan
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引用次数: 0

Abstract

Lymphocyte activation is critical in regulating immune responses. The resulting T-cell proliferation has been implicated in the pathogenesis of a variety of autoimmune diseases, such as SLE and rheumatoid arthritis. ConA (concanavalin A)-induced activation has been widely used in the T lymphocytes model of immune-mediated liver injury, autoimmune hepatitis, and so on. In those works, it usually requires fluorescent labeling or cell staining to confirm whether the cells are transformed successfully after medicine treatment to figure out efficacy/pharmacology. The detection preparation steps are time-consuming and have limitations for further proteomic/genomic identifications. Here, a label-free microfluidic method is established to detect lymphocyte activation degree. The lymphocyte and ConA-activated lymphocyte were investigated by a microfluidic device. According to where single cells in the sample were captured in the designed channel, lymphocyte and ConA-activated samples are differentiated and characterized by population electric field factors, 2.08 × 104 and 2.21 × 104 V/m, respectively. Furthermore, salidroside, a herbal medicine that was documented to promote the transformation, was used to treat lymphocyte cells, and the treated cell population is detected to be 2.67 × 104 V/m. The characterization indicates an increasing trend with the activation degree. The result maintains a high consistency with traditional staining methods with transformed cells of 15.8%, 28.8%, and 48.3% in each cell population. Dielectrophoresis is promising to work as a tool for detecting lymphocyte transformation and medical efficacy detection.

利用微流体技术在单细胞水平对 ConA 诱导的 T 淋巴细胞活化进行无标记检测。
淋巴细胞活化是调节免疫反应的关键。由此导致的 T 细胞增殖与系统性红斑狼疮和类风湿性关节炎等多种自身免疫性疾病的发病机制有关。ConA(concanavalin A)诱导的活化已被广泛应用于免疫介导的肝损伤、自身免疫性肝炎等的 T 淋巴细胞模型中。在这些工作中,通常需要通过荧光标记或细胞染色来确认药物治疗后细胞是否成功转化,以了解疗效/药理。这些检测准备步骤不仅耗时,而且对进一步的蛋白质组/基因组鉴定也有限制。本文建立了一种检测淋巴细胞活化程度的无标记微流控方法。淋巴细胞和 ConA 激活的淋巴细胞由微流控装置进行检测。根据样品中单细胞在设计通道中被捕获的位置,淋巴细胞和 ConA 激活的样品被区分开来,并分别以 2.08 × 104 V/m 和 2.21 × 104 V/m 的群体电场因子为特征。此外,用一种据记载能促进转化的中药--水杨甙来处理淋巴细胞,检测到处理后的细胞群电场系数为 2.67 × 104 V/m。表征结果表明,活化程度呈上升趋势。结果与传统染色方法保持高度一致,每个细胞群中转化细胞的比例分别为 15.8%、28.8% 和 48.3%。压电效应有望成为检测淋巴细胞转化和医疗效果检测的一种工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
ELECTROPHORESIS
ELECTROPHORESIS 生物-分析化学
CiteScore
6.30
自引率
13.80%
发文量
244
审稿时长
1.9 months
期刊介绍: ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.). Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences. Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases. Papers describing the application of standard electrophoretic methods will not be considered. Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics: • Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry • Single cell and subcellular analysis • Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS) • Nanoscale/nanopore DNA sequencing (next generation sequencing) • Micro- and nanoscale sample preparation • Nanoparticles and cells analyses by dielectrophoresis • Separation-based analysis using nanoparticles, nanotubes and nanowires.
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