Department of Bio-medical Sciences, University of Catania, Via Androne 83, 95124 Catania, Italy.
Inflamm Res. 2012 Oct;61(10):1131-9. doi: 10.1007/s00011-012-0506-y. Epub 2012 Jun 16.
To evaluate the heat shock protein (HSP) variation during the differentiation and polarization of human macrophages.
Gene expression analysis was investigated by real-time PCR from mRNA of human monocytes obtained from the buffy coats of healthy volunteers, polarized to classically activated macrophages (or M1), whose prototypical activating stimuli are interferon-gamma and lipopolysaccharide, and alternatively activated macrophages (or M2) obtained by interleukin-4 exposure. The modulation of HSPs at the transcriptomic levels was investigated using oligonucleotide microarray in the process of primary human monocyte-to-macrophage maturation and subsequent polarization into M1 or M2 cells.
We found that 11 HSPs transcripts were modulated throughout monocyte-to-macrophage differentiation. Furthermore a considerable effect on HSP expression was detected in conjunction with the M1 polarizing condition. This affected 21 transcripts in M1 cells, with 6 of them significantly upregulated in comparison to unpolarized macrophages, whereas 15 were downregulated. Slight changes in HSPs expression were observed in M2 cells when compared to unpolarized macrophages. Under these circumstances only five transcripts were significantly modulated. Interestingly, HSPBAP1 was the only HSP significantly downregulated in both M1 and M2 conditions parallel to a significant up-regulation of its target HSPB1.
Our study revealed that monocytes undergoing maturation differentially regulate the expression of several members of HSPs and that distinct patterns of HSP expression characterize the M1 and M2 effector stages of macrophage life.
评估人巨噬细胞分化和极化过程中的热休克蛋白（HSP）变化。
通过实时 PCR 从来自健康志愿者的外周血单个核细胞（Buffy coat）中的人单核细胞的 mRNA 中进行基因表达分析，将其极化为经典激活的巨噬细胞（或 M1），其典型激活刺激物是干扰素-γ和脂多糖，以及通过白细胞介素-4 暴露获得的替代性激活的巨噬细胞（或 M2）。在原代人单核细胞向巨噬细胞成熟以及随后向 M1 或 M2 细胞极化的过程中，使用寡核苷酸微阵列研究 HSP 在转录组水平上的调节。
我们发现 11 个 HSP 转录本在单核细胞向巨噬细胞分化过程中被调节。此外，在 M1 极化条件下，HSP 表达也受到了相当大的影响。这影响了 M1 细胞中的 21 个转录本，其中 6 个与未极化的巨噬细胞相比显著上调，而 15 个下调。与未极化的巨噬细胞相比，在 M2 细胞中观察到 HSPs 表达的轻微变化。在这种情况下，只有 5 个转录本被显著调节。有趣的是，HSPBAP1 是唯一在 M1 和 M2 条件下均显著下调的 HSP，同时其靶标 HSPB1 显著上调。
我们的研究表明，成熟中的单核细胞差异调节 HSPs 的几个成员的表达，并且 HSP 表达的不同模式特征化巨噬细胞生命的 M1 和 M2 效应阶段。
