Unraveling Macrophage Heterogeneity in Erythroblastic Islands.
Unraveling Macrophage Heterogeneity in Erythroblastic Islands.
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揭示成红细胞岛中巨噬细胞的异质性。
DOI：
10.3389/fimmu.2017.01140
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发表时间：
2017
影响因子：
7.3
通讯作者：
Kalfa TA
中科院分区：
文献类型：
--
作者：
Seu KG;Papoin J;Fessler R;Hom J;Huang G;Mohandas N;Blanc L;Kalfa TA
哺乳动物红细胞生成发生在成红细胞岛（EBI）内，成熟的成红细胞与中央巨噬细胞密切相互作用。虽然普遍认为EBI巨噬细胞在红细胞生成中起重要作用，但对它们支持红细胞生成的机制的彻底研究在很大程度上受到无法鉴定和分离构成EBI的特定巨噬细胞亚群的限制。早期的研究利用免疫组织化学或免疫荧光来研究EBI的形态和结构，而最近的努力已经使用流式细胞术对EBI及其中央巨噬细胞进行高通量定量表征。然而，这些基于EBI巨噬细胞是同质群体（例如F4/80+/CD 169 +/VCAM-1+）的预期的方法提供了不完整的图片，并且可能忽略了关于岛及其中央巨噬细胞的性质和生物学的关键信息。在这里，我们提出了一种新的方法，用于分析EBI巨噬细胞从造血组织的小鼠和大鼠使用多光谱成像流式细胞术（IFC），它结合了高通量的优势，流式细胞术的形态和荧光特征来自显微镜。该方法提供了EBI的定量分析，以及中央巨噬细胞和相关细胞的结构和形态学细节。重要的是，结合定量软件特征的图像可用于评估表型标志物的共表达，这是至关重要的，因为用于鉴定巨噬细胞的一些抗原（例如，F4/80和CD 11b）可以在与岛相关的非红系细胞上表达，而不是或除了中央巨噬细胞本身之外。我们已经使用这种方法来分析来自不同造血组织的天然EBI，并评估了先前报道在EBI巨噬细胞上表达的几种标记物的表达。我们发现，VCAM-1，F4/80和CD 169在EBI内的中央巨噬细胞中不均匀表达，而CD 11b，尽管在岛内的细胞中大量表达，但在EBI巨噬细胞上不表达。此外，与小鼠相比，大鼠中EBI表型的差异表明这些物种之间存在潜在的功能差异。这些数据证明了IFC在EBI的分析和表征中的有用性，更重要的是在探索EBI巨噬细胞的异质性和可塑性中的有用性。
Mammalian erythropoiesis occurs within erythroblastic islands (EBIs), niches where maturing erythroblasts interact closely with a central macrophage. While it is generally accepted that EBI macrophages play an important role in erythropoiesis, thorough investigation of the mechanisms by which they support erythropoiesis is limited largely by inability to identify and isolate the specific macrophage sub-population that constitute the EBI. Early studies utilized immunohistochemistry or immunofluorescence to study EBI morphology and structure, while more recent efforts have used flow cytometry for high-throughput quantitative characterization of EBIs and their central macrophages. However, these approaches based on the expectation that EBI macrophages are a homogeneous population (F4/80+/CD169+/VCAM-1+ for example) provide an incomplete picture and potentially overlook critical information about the nature and biology of the islands and their central macrophages. Here, we present a novel method for analysis of EBI macrophages from hematopoietic tissues of mice and rats using multispectral imaging flow cytometry (IFC), which combines the high-throughput advantage of flow cytometry with the morphological and fluorescence features derived from microscopy. This method provides both quantitative analysis of EBIs, as well as structural and morphological details of the central macrophages and associated cells. Importantly, the images, combined with quantitative software features, can be used to evaluate co-expression of phenotypic markers which is crucial since some antigens used to identify macrophages (e.g., F4/80 and CD11b) can be expressed on non-erythroid cells associated with the islands instead of, or in addition to the central macrophage itself. We have used this method to analyze native EBIs from different hematopoietic tissues and evaluated the expression of several markers that have been previously reported to be expressed on EBI macrophages. We found that VCAM-1, F4/80, and CD169 are expressed heterogeneously by the central macrophages within the EBIs, while CD11b, although abundantly expressed by cells within the islands, is not expressed on the EBI macrophages. Moreover, differences in the phenotype of EBIs in rats compared to mice point to potential functional differences between these species. These data demonstrate the usefulness of IFC in analysis and characterization of EBIs and more importantly in exploring the heterogeneity and plasticity of EBI macrophages.
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影响因子：
5.4
作者：
Gordon, Siamon;Hamann, Joerg;Stacey, Martin
通讯作者：
Stacey, Martin
影响因子：
82.9
作者：
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--
影响因子：
20.3
作者：
Korolnek, Tamara;Hamza, Iqbal
通讯作者：
Hamza, Iqbal
影响因子：
20.3
作者：
Konstantinidis, Diamantis G.;Pushkaran, Suvarnamala;Kalfa, Theodosia A.
通讯作者：
Kalfa, Theodosia A.
影响因子：
--
作者：
Giger KM;Kalfa TA
通讯作者：
Kalfa TA