Expression data from healthy and malignant (chronic lymphocytic leukemia, CLL) human B-lymphocytes after B-cell receptor stimulation

Three different cell populations (6 healthy B-lymphocytes, 6 leukemic CLL B-lymphocyte of indolent form and 5 leukemic CLL B-lymphocyte of aggressive form) were stimulated in vitro with an anti-IgM antibody, activating the B-cell receptor (BCR). We analyzed the gene expression at 4 time points (60, 90, 210 and 390 minutes). Each gene expression measurement is performed both in stimulated cells and in control unstimulated cells. For one aggressive CLL case, we silenced expression of DUSP1 by transfecting DUSP1-specific RNAi and, as a control, transfected cells with a non-targeting RNAi. We then stimulated the BCR of these cells and analyzed the gene expression at the same time points in stimulated cells and in control unstimulated cells. B-cells were negatively selected from healthy donors and previously untreated CLL patients. BCR stimulated and unstimulated control B-cells were treated at four time points after stimulation for total RNA extraction and hybridization on Affymetrix microarrays.

本数据集涵盖三组不同的细胞群：6例健康B淋巴细胞、6例惰性亚型慢性淋巴细胞白血病（Chronic Lymphocytic Leukemia, CLL）B淋巴细胞，以及5例侵袭性亚型慢性淋巴细胞白血病（Chronic Lymphocytic Leukemia, CLL）B淋巴细胞。研究人员将上述细胞在体外以抗IgM抗体刺激，以激活B细胞受体（B-cell receptor, BCR）。我们在60、90、210及390分钟四个时间点对基因表达水平进行检测。每项基因表达分析均同时在刺激组细胞与未刺激的对照组细胞中开展。针对1例侵袭性CLL病例，我们通过转染DUSP1特异性RNA干扰（RNA interference, RNAi）沉默DUSP1的基因表达，并以转染非靶向RNA干扰的细胞作为对照。随后我们对这些细胞的B细胞受体进行刺激，并在上述相同时间点分别分析刺激组与未刺激对照组细胞的基因表达。B细胞均从健康捐献者及此前未接受治疗的CLL患者体内通过阴性分选法分离获得。BCR刺激组及未刺激对照组B细胞在刺激后的四个时间点接受处理，用于总RNA提取，并在Affymetrix基因芯片上完成杂交。