Morphologic, phenotypic, and transcriptomic characterization of classically and alternatively activated canine blood-derived macrophages in vitro.
Morphologic, phenotypic, and transcriptomic characterization of classically and alternatively activated canine blood-derived macrophages in vitro.
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DOI：
10.1371/journal.pone.0183572
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Spitzbarth I
中科院分区：
文献类型：
--
作者：
Heinrich F;Lehmbecker A;Raddatz BB;Kegler K;Tipold A;Stein VM;Kalkuhl A;Deschl U;Baumgärtner W;Ulrich R;Spitzbarth I
巨噬细胞是在组织稳态和炎症中起关键作用的异质细胞群体，并且它们的表型强烈依赖于微环境。尽管其作为人类疾病的转化动物模型的重要性日益增加，但关于犬中巨噬细胞极化的知识存在相当大的差距。本研究在体外对未刺激（M0）、M1-（GM-CSF、LPS、IFNγ刺激）和M2-（M-CSF、IL-4刺激）极化的犬血液源性巨噬细胞的形态学、表型和转录组学特征进行了全面研究。扫描电子显微镜显示极化的巨噬细胞的不同形态，在M2-巨噬细胞中形成多核细胞，而免疫荧光采用基于文献的原型抗体抗CD 16，CD 32，iNOS，MHC II类（M1-标志物）、CD 163、CD 206和CD 206酶-1（M2-标志物）表明，只有CD 206能够区分M2-巨噬细胞与其他两种表型，突出了该分子作为犬M2-巨噬细胞的有希望的标志物。全球微阵列分析揭示了极化犬巨噬细胞转录组的深刻变化。功能分析指出，M1-极化与生物过程，如“呼吸爆发”，而M2-极化与过程，如“有丝分裂”。基于文献的标记基因选择显示，与鼠和人巨噬细胞的原型标记相比，狗的基因组中只有微小的重叠。使用监督聚类的生物标志物选择表明，乳胶蛋白（LXN）和跨膜4-域，亚家族A，成员2（MS 4A 2）分别是犬M1-和M2-巨噬细胞最强大的预测生物标志物。两种标记物的免疫荧光表明，这两种蛋白质的表达，巨噬细胞在体外，但未能揭示犬M1和M2-巨噬细胞之间的差异。目前的研究提供了一个坚实的基础，为今后的研究中的作用，巨噬细胞极化的自发性疾病的狗，一个物种，具有新兴的重要性的转化研究。
Macrophages are a heterogeneous cell population playing a pivotal role in tissue homeostasis and inflammation, and their phenotype strongly depends on the micromilieu. Despite its increasing importance as a translational animal model for human diseases, there is a considerable gap of knowledge with respect to macrophage polarization in dogs. The present study comprehensively investigated the morphologic, phenotypic, and transcriptomic characteristics of unstimulated (M0), M1- (GM-CSF, LPS, IFNγ-stimulated) and M2- (M-CSF, IL-4-stimulated)-polarized canine blood-derived macrophages in vitro. Scanning electron microscopy revealed distinct morphologies of polarized macrophages with formation of multinucleated cells in M2-macrophages, while immunofluorescence employing literature-based prototype-antibodies against CD16, CD32, iNOS, MHC class II (M1-markers), CD163, CD206, and arginase-1 (M2-markers) demonstrated that only CD206 was able to discriminate M2-macrophages from both other phenotypes, highlighting this molecule as a promising marker for canine M2-macrophages. Global microarray analysis revealed profound changes in the transcriptome of polarized canine macrophages. Functional analysis pointed out that M1-polarization was associated with biological processes such as “respiratory burst”, whereas M2-polarization was associated with processes such as “mitosis”. Literature-based marker gene selection revealed only minor overlaps in the gene sets of the dog compared to prototype markers of murine and human macrophages. Biomarker selection using supervised clustering suggested latexin (LXN) and membrane-spanning 4-domains, subfamily A, member 2 (MS4A2) to be the most powerful predicting biomarkers for canine M1- and M2-macrophages, respectively. Immunofluorescence for both markers demonstrated expression of both proteins by macrophages in vitro but failed to reveal differences between canine M1 and M2-macrophages. The present study provides a solid basis for future studies upon the role of macrophage polarization in spontaneous diseases of the dog, a species that has emerging importance for translational research.
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影响因子：
--
作者：
Estko M;Baumgartner S;Urech K;Kunz M;Regueiro U;Heusser P;Weissenstein U
通讯作者：
Weissenstein U
影响因子：
3.7
作者：
Derlindati E;Dei Cas A;Montanini B;Spigoni V;Curella V;Aldigeri R;Ardigò D;Zavaroni I;Bonadonna RC
通讯作者：
Bonadonna RC
影响因子：
5.7
作者：
Aagaard, A;Listwan, P;Martin, JL
通讯作者：
Martin, JL
影响因子：
6.2
作者：
Durafourt, Bryce A.;Moore, Craig S.;Antel, Jack P.
通讯作者：
Antel, Jack P.
影响因子：
29
作者：
Bouhlel, M. Amine;Derudas, Bruno;Chinetti-Gbaguidi, Giulia
通讯作者：
Chinetti-Gbaguidi, Giulia