基于微流控芯片快速高分辨率检测亚群识别RAW 264.7免疫耐受细胞和筛选药物的新方法

A DC-iDEP-based fast and high-resolution method for detection of LPS tolerance of RAW264.7 macrophages and screening of therapeutic agents

摘要:

脓毒症是重症医学中一种常见且致死率高的疾病，是早期过度炎症转变为长期免疫抑制的过程，在先天免疫细胞中表现为细胞应激性降低，即脂多糖耐受性。现有大多数检测细胞脂多糖耐受的方法，无法准确识别少量存在的亚群细胞，且面对复杂的细胞体系鉴定能力有限，亟待开发一种快速、无标记的检测免疫耐受细胞状态的方法。本研究应用微流控直流介电泳芯片通过生物物理性质对炎症细胞、免疫耐受细胞状态进行区分性的表征，并尝试通过改变细胞生物物理性质筛选逆转免疫耐受的药物。结果表明，三种细胞的生物物理特征值分别为4.28×10⁸、3.13×10⁸和4.25×10⁸ V/m²，即所建立的方法可用于区分LPS耐受细胞。本芯片有望应用于医学诊断和药物筛查。

Abstract:

Sepsis is a leading life-threatening problem in intensive care medicine. The recent studies have given insights into the transition from inflammation to long-term immunosuppression in sepsis. This condition might cause physiological changes that comprise the lipopolysaccharide (LPS) tolerance. Most studies about the LPS tolerance focus on the reduced ability of macrophages to secrete pro-inflammatory cytokines. Although this method has identified various molecular changes, it remains ambiguous since changes in the whole cell population are measured as an average and markers are required for cell recognition. A fast and label-free method is in demand to detect cell tolerance and screen therapeutic agents that might reverse the process. In this study, direct current insulator-based dielectrophoresis (DC-iDEP) was used to characterize the biophysical properties (EKMr) of inflamed cells, LPS-tolerant cells, and cells treated with therapeutic agents. The results showed that the EKMr of these cells was 4.28×10⁸, 3.13×10⁸, and 4.25×10⁸ V/m², respectively, suggesting that the established method was useful in distinguishing LPS-tolerant cells. The device holds the promise to be applied in medical diagnosis and medicine screening.