用于固定全血样本免疫表型表征的高维光谱流式细胞术检测panel

摘要(中文译文)

外周血单个核细胞（PBMCs）常用于临床研究中的免疫分析，但其分离和冷冻保存存在后勤方面的挑战，尤其是在研究设施可及性有限的环境中。为解决这一问题，我们实施了一种全血固定方案，以保存表面蛋白表位，并使高维流式细胞术无需即时样本处理即可进行。我们开发并优化了36标志物淋系和35标志物髓系光谱流式细胞术panel，用于表征多种免疫细胞群体，包括T细胞亚群、B细胞、NK细胞、单核细胞、树突状细胞和粒细胞。Panel设计重点关注抗体克隆与固定抗原的兼容性、尽量减少非特异性结合以及可靠检测趋化因子受体。通过比较新鲜与固定全血评估性能，显示主要免疫谱系的鉴定结果一致。通过使用来自健康供者的冷冻保存固定血液历时两年半，证明了长期稳定性，免疫图谱保持稳定。这些发现支持将固定全血用于高维光谱流式细胞术及其在纵向临床免疫监测中的实用性。

Abstract

Peripheral blood mononuclear cells (PBMCs) are commonly used for immune profiling in clinical research, but their isolation and cryopreservation present logistical challenges, particularly in settings with limited access to research facilities. To address this, we implemented a whole blood fixation protocol to preserve surface protein epitopes and enable high-dimensional flow cytometry without the need for immediate sample processing. A 36-marker lymphoid and 35-marker myeloid spectral flow cytometry panels were developed and optimized to characterize diverse immune populations, including T cell subsets, B cells, NK cells, monocytes, dendritic cells, and granulocytes. Panel design focused on antibody clone compatibility with fixed antigens, minimizing non-specific binding, and reliably detecting chemokine receptors. Performance was evaluated by comparing fresh and fixed whole blood, revealing consistent identification of major immune lineages. Long-term stability was demonstrated by using cryopreserved fixed blood from a healthy donor over two and half years, with immune profiles remaining stable. These findings support the use of fixed whole blood for high-dimensional spectral cytometry and its utility in longitudinal clinical immune monitoring.

如何引用

Cytometry. Part A : the journal of the International Society for Analytical Cytology 的其他文献

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