Comprehensive assessment of chemokine expression profiles by flow cytometry
Comprehensive assessment of chemokine expression profiles by flow cytometry
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DOI：
10.1172/jci40645
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发表时间：
2010-03-01
影响因子：
15.9
通讯作者：
Homann, Dirk
中科院分区：
文献类型：
--
作者：
Eberlein, Jens;Nguyen, Tom T.;Homann, Dirk
趋化因子是一个大家族，主要分泌的分子参与许多生理和病理生理过程的调节。尽管多年的研究，大多数趋化因子的精确细胞来源仍然不完全定义的结果，有限的可用性合适的试剂，以可视化在单细胞水平的趋化因子蛋白质的表达。在这里，我们开发了一个简单的流式细胞术为基础的分析，使用市售的趋化因子特异性抗体的有效细胞相关检测的39种小鼠趋化因子的37。为了证明这种方法的实用性，我们用它来重新评估的性质，稳态趋化因子在造血隔室，描绘完整的趋化因子谱的NK细胞和B细胞在响应主要的多克隆刺激，并评估趋化因子反应的DC细菌感染。这种分析方法的多功能性进一步证明了其应用于选定的人类趋化因子，并应大大促进任何未来的调查趋化因子生物学在大。
The chemokines are a large family of mainly secreted molecules involved in the regulation of numerous physiological and pathophysiological processes. Despite many years of investigation, the precise cellular sources of most chemokines have remained incompletely defined as a consequence of the limited availability of suitable reagents to visualize the expression of chemokine proteins at the single-cell level. Here, we developed a simple flow cytometry-based assay using commercially available chemokine-specific antibodies for efficient cell-associated detection of 37 of 39 murine chemokines. To demonstrate the utility of this methodology, we used it to reevaluate the nature of homeostatic chemokines in the hematopoietic compartment, to delineate the complete chemokine profiles of NK cells and B cells in response to major polyclonal stimuli, and to assess the chemokine response of DCs to bacterial infection. The versatility of this analytical methodology was further demonstrated by its application to selected human chemokines and should greatly facilitate any future investigation into chemokine biology at large.