Phenotypic and functional characterization of macrophages with therapeutic potential generated from human cirrhotic monocytes in a cohort study.
Phenotypic and functional characterization of macrophages with therapeutic potential generated from human cirrhotic monocytes in a cohort study.
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DOI：
10.1016/j.jcyt.2015.07.016
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发表时间：
2015-11
期刊：
影响因子：
4.5
通讯作者：
Forbes SJ
中科院分区：
文献类型：
--
作者：
Moore JK;Mackinnon AC;Wojtacha D;Pope C;Fraser AR;Burgoyne P;Bailey L;Pass C;Atkinson A;Mcgowan NW;Manson L;Turner ML;Campbell JD;Forbes SJ
巨噬细胞在肝脏中具有复杂的作用。本研究的目的是比较对照组和肝硬化患者之间人单核细胞衍生的巨噬细胞的概况，以确定慢性炎症是否影响前体数量或表型，最终目的是开发肝硬化的细胞疗法。在鼠肝纤维化模型中输注人巨噬细胞显示肝损伤标志物（丙氨酸转氨酶、胆红素、天冬氨酸转氨酶）和纤维化标志物（转化生长因子-β、α-平滑肌肌动蛋白、磷脂酰丝氨酸受体）降低，肝再生标志物（基质金属蛋白酶[MMP]-9，MMP-12和TNF相关弱凋亡诱导物）增加。然后从对照中分离CD 14+单核细胞。使用药品生产质量管理规范兼容技术，将单核细胞成熟为巨噬细胞7天。从血小板减少患者（n = 9）和对照组（n = 10）中分离的CD 14+单核细胞的平均数量之间无显著差异;分别为2.8 ± SEM 0.54 × 108和2.5 ± 0.56 × 108。成熟巨噬细胞培养的平均产量在肺炎患者和对照组之间也没有显著差异（分别为0.9 × 108 ± 0.38 × 108，存活率超过90%和0.65 × 108 ± 0.16 × 108。成熟为巨噬细胞导致许多基因（MMP-9、CCL 2、白细胞介素[IL]-10和TNF相关的弱凋亡诱导物）上调。一个细胞因子和趋化因子聚合酶链反应阵列，比较对照组和炎性巨噬细胞，显示没有统计学上的显着差异。巨噬细胞可以从血小板减少患者的血小板来源的CD 14单核细胞中分化出来，并形成与对照巨噬细胞相同的促消退表型，表明它们适用于临床治疗。
Macrophages have complex roles in the liver. The aim of this study was to compare profiles of human monocyte-derived macrophages between controls and cirrhotic patients, to determine whether chronic inflammation affects precursor number or the phenotype, with the eventual aim to develop a cell therapy for cirrhosis. Infusion of human macrophages in a murine liver fibrosis model demonstrated a decrease in markers of liver injury (alanine transaminase, bilirubin, aspartate transaminase) and fibrosis (transforming growth factor-β, α-smooth muscle actin, phosphatidylserine receptor) and an increase in markers of liver regeneration (matrix metalloproteinases [MMP]-9, MMP-12 and TNF-related weak inducer of apoptosis). CD14+ monocytes were then isolated from controls. Monocytes were matured into macrophages for 7 days using a Good Manufacturing Practice–compatible technique. There was no significant difference between the mean number of CD14+ monocytes isolated from cirrhotic patients (n = 9) and controls (n = 10); 2.8 ± SEM 0.54 × 108 and 2.5 ± 0.56 × 108, respectively. The mean yield of mature macrophages cultured was also not significantly different between cirrhotic patients and controls (0.9 × 108 ± 0.38 × 108, with more than 90% viability and 0.65 × 108 ± 0.16 × 108, respectively. Maturation to macrophages resulted in up-regulation of a number of genes (MMP-9, CCL2, interleukin [IL]-10 and TNF-related weak inducer of apoptosis). A cytokine and chemokine polymerase chain reaction array, comparing the control and cirrhotic macrophages, revealed no statistically significant differences. Macrophages can be differentiated from cirrhotic patients' apheresis-derived CD14 monocytes and develop the same pro-resolution phenotype as control macrophages, indicating their suitability for clinical therapy.