Expression data from human macrophages

Human CD14 positive monocytes were purified from healthy volunteers’ blood and cultured in vitro for 4, 12, 24, 72 hours. While culturing, macrophages were activated alternatively with interleukin-4 (IL-4 100 ng/ml) or classically with interferon-gamma (IFNg 100 ng/ml)+tumor necrosis factor (TNF 50 ng/ml) or left without activation. Simultaneously, macrophages were also treated with vehicle (DMSO:ethanol) or 1mM synthetic PPARg agonist, Rosiglitazone. We used Affymetrix microarrays (U133Plus 2.0) to analyze activation and PPARg-induced gene expression changes. Monocytes from 3 donors were used and treated as indicated in Summary.

本研究从健康志愿者血液中纯化人CD14阳性单核细胞（human CD14-positive monocytes），并进行体外培养，培养时长分别设置为4、12、24、72小时。培养期间，对巨噬细胞分别采用替代性激活方案（添加100 ng/ml白细胞介素-4（IL-4））、经典激活方案（联合添加100 ng/ml γ干扰素（IFN-γ）与50 ng/ml肿瘤坏死因子（TNF）），或不进行任何激活处理作为对照。同时，部分巨噬细胞采用溶剂对照（二甲基亚砜:乙醇，DMSO:ethanol）处理，或加入1 mM合成型过氧化物酶体增殖物激活受体γ（PPARγ）激动剂罗格列酮（Rosiglitazone）进行干预。本研究采用Affymetrix公司的U133Plus 2.0基因芯片，分析巨噬细胞激活状态及PPARγ介导的基因表达变化情况。本实验共使用3名健康志愿者的单核细胞，具体处理方案详见实验摘要。