Gene expression analysis illuminates the transcriptional programs underlying the functional activity of ex vivo-expanded granulocytes
Gene expression analysis illuminates the transcriptional programs underlying the functional activity of ex vivo-expanded granulocytes
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DOI：
10.1152/physiolgenomics.00053.2007
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发表时间：
2007-09-19
影响因子：
4.6
通讯作者：
Miller, William M.
中科院分区：
文献类型：
--
作者：
Huang, Li Ting;Paredes, Carlos J.;Miller, William M.
全球基因表达分析建立了体外扩增(EXE)人粒细胞功能活性发展的时间表达模式和程序，以及与外周血(PB)粒细胞的差异。CD34(+)祖细胞培养3wk诱导快速扩增和粒细胞分化，第3天CD15(+)细胞达到40%，第12天达到90%。吞噬和呼吸爆发活性随着CD15(+)(+)CD11b(+)细胞比例的增加而增加，并在第17天达到高峰。然而，CD15(+)(+)CD11b(+)细胞只有25%吞噬，呼吸爆发活性是外周血粒细胞的1/3。晚期EXE粒细胞和PB粒细胞的Fcγ受体(-1A、-2A、-2C、-3A)和补体受体(-1、-3、-4)的表达相似。后来CD36(由巨噬细胞表达)的下调表明粒细胞分化早期的谱系可塑性。在成熟的EXE和PB粒细胞中，大多数Fcγ受体介导的吞噬信号蛋白的表达相似，包括高水平表达HCK、FGR和肌动蛋白相关蛋白2/3复合体。EXE粒细胞Lyn、CDC42、pleckstrin和PKCβ(I)的低表达可能是吞噬功能降低的原因。外周血粒细胞和成熟EXE粒细胞表达相似水平的NADPH氧化酶复合体基因和fMLP介导的呼吸爆发受体。EXE粒细胞爆裂活性降低可能与Raf1和PKC Zeta的低表达有关。成熟EXE和外周血粒细胞中Toll样受体(TLR2)2、TLR1和CD14的高表达支持TLR2和CD14途径在酵母多糖介导的呼吸爆发活动中的作用。EXE粒细胞活性降低可能是由于抑制TLR介导的信号转导的IRAK3表达增加所致。
Global gene expression analysis established the temporal expression patterns and programs underlying the development of functional activity of ex vivo-expanded (EXE) human granulocytes, as well as differences compared with peripheral blood (PB) granulocytes. CD34(+) progenitor cells were cultured for 3 wk to induce rapid expansion and granulocytic differentiation, with 40% CD15(+) cells by day 3 and 90% by day 12. Phagocytic and respiratory burst activity increased with the fraction of CD15(+) (+)CD11b(+) cells (myelocytes to segmented) and peaked by day 17. However, only 25% of CD15(+) (+)CD11b(+) cells were phagocytic, and respiratory burst activity was one-third that of PB granulocytes. EXE granulocytes from later days and PB granulocytes showed similar expression of Fc gamma receptors (-1A, -2A, -2C, -3A) and complement receptors (-1, -3, -4). Later downregulation of CD36 (expressed by macrophages) suggests lineage plasticity early in granulocytic differentiation. Expression in mature EXE and PB granulocytes was similar for most Fc gamma receptor-mediated phagocytosis signaling proteins, including high-level expression of Hck, Fgr, and the actin-related protein 2/3 complex. Lower expression of Lyn, Cdc42, pleckstrin, and PKC beta(I) by EXE granulocytes may explain decreased phagocytosis. PB and mature EXE granulocytes expressed similar levels of NADPH oxidase complex genes and receptors for fMLP-mediated respiratory burst. Lower burst activity by EXE granulocytes may result from lower expression of Raf1 and PKC zeta. Elevated expression of toll-like receptor (TLR)2, TLR1, and CD14 in mature EXE and PB granulocytes supports a role for the TLR2 and CD14 pathway in zymosan-mediated respiratory burst activity. Lower activity in EXE granulocytes may be due to greater expression of IRAK3, which inhibits TLR-mediated signaling.