The identification of markers of macrophage differentiation in PMA-stimulated THP-1 cells and monocyte-derived macrophages.
The identification of markers of macrophage differentiation in PMA-stimulated THP-1 cells and monocyte-derived macrophages.
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DOI：
10.1371/journal.pone.0008668
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发表时间：
2010-01-13
期刊：
影响因子：
3.7
通讯作者：
Dockrell DH
中科院分区：
文献类型：
--
作者：
Daigneault M;Preston JA;Marriott HM;Whyte MK;Dockrell DH
分化巨噬细胞是机体固有免疫应答的常驻组织吞噬细胞和前哨细胞。成熟组织巨噬细胞的表型是环境依赖性和分化依赖性印迹的复合表型。phorboll -12-肉豆酸酯-13-乙酸酯（PMA）和1,25-二羟基维生素D3 （VD3）是单核细胞系中常用的诱导巨噬细胞分化的刺激，但与原代组织巨噬细胞相比，其分化程度尚不清楚。我们比较了利用VD3和PMA不同分化方案后的原单核细胞THP-1细胞系与原代人单核细胞或单核细胞源性巨噬细胞（MDM）的表型。这两种刺激都诱导了细胞形态的变化，但都没有表现出与MDM相当的分化。相比之下，PMA处理后在没有PMA的培养中休息5天（PMAr）增加了细胞质与核的比率，增加了线粒体和溶酶体的数量，并改变了分化依赖的细胞表面标记物，其模式与MDM相似。此外，PMAr细胞对凋亡刺激表现出相对的抗性，并维持与MDM相似的分化依赖性抗凋亡蛋白Mcl-1的水平。PMAr细胞保留了对乳胶珠的高吞噬能力，并表达了类似MDM对TLR配体的反应的细胞因子谱，特别是对TLR2的明显反应。此外，MDM和PMAr对刺激定向极化均保持了显著的可塑性。这些发现表明，改良的PMA分化方案可以增强THP-1细胞的巨噬细胞分化，并将线粒体和溶酶体数量的增加、对凋亡的抗性和TLR2反应的效力作为转化细胞巨噬细胞分化水平的重要鉴别因子。
Differentiated macrophages are the resident tissue phagocytes and sentinel cells of the innate immune response. The phenotype of mature tissue macrophages represents the composite of environmental and differentiation-dependent imprinting. Phorbol-12-myristate-13-acetate (PMA) and 1,25-dihydroxyvitamin D3 (VD3) are stimuli commonly used to induce macrophage differentiation in monocytic cell lines but the extent of differentiation in comparison to primary tissue macrophages is unclear. We have compared the phenotype of the promonocytic THP-1 cell line after various protocols of differentiation utilising VD3 and PMA in comparison to primary human monocytes or monocyte-derived macrophages (MDM). Both stimuli induced changes in cell morphology indicative of differentiation but neither showed differentiation comparable to MDM. In contrast, PMA treatment followed by 5 days resting in culture without PMA (PMAr) increased cytoplasmic to nuclear ratio, increased mitochondrial and lysosomal numbers and altered differentiation-dependent cell surface markers in a pattern similar to MDM. Moreover, PMAr cells showed relative resistance to apoptotic stimuli and maintained levels of the differentiation-dependent anti-apoptotic protein Mcl-1 similar to MDM. PMAr cells retained a high phagocytic capacity for latex beads, and expressed a cytokine profile that resembled MDM in response to TLR ligands, in particular with marked TLR2 responses. Moreover, both MDM and PMAr retained marked plasticity to stimulus-directed polarization. These findings suggest a modified PMA differentiation protocol can enhance macrophage differentiation of THP-1 cells and identify increased numbers of mitochondria and lysosomes, resistance to apoptosis and the potency of TLR2 responses as important discriminators of the level of macrophage differentiation for transformed cells.