LMP1+SLAMF1high cells are associated with drug resistance in Epstein-Barr virus-positive Farage cells. Homo sapiens

We isolated Farage cells expressing the cell surface marker SLAMF1. LMP1 and its target gene CCL22 were highly expressed in SLAMF1high Farage cells. These cells survived longer following treatment with a combination of cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP). Genes associated with interferon-alpha, allograft rejection, NF-kB, STAT3 and 5 were also overexpressed in the surviving Farage cells. Overall design: The three experimental conditions were designed: 1) Farage cells were sorted according to expression level of SLAMF1 and labeled as SLAM_high or SLAM_low.; 2) Farage cells treated with 1 μg/ml CHOP for 2 days and 3 days compared with untreated Farage cells; 3) Farage cells incubated with 1 μg/ml CHOP plus 50 ng/ml IL4 compared with 50 ng/ml IL4 alone for 2 days. Genome-wide analysis of all cell lines were performed using the Affymetrix HuGene ST 2.0 Array Platform. Genes with differential expression were analyzed using Statistical Analysis of Microarrays (SAM) software, and a signature of genes determined.

我们分离了表达细胞表面标志物信号淋巴细胞活化分子家族成员1（SLAMF1）的Farage细胞。在SLAMF1高表达的Farage细胞中，潜伏膜蛋白1（LMP1）及其靶基因趋化因子配体22（CCL22）呈高表达水平。经环磷酰胺、多柔比星、长春新碱、泼尼松（CHOP）联合方案处理后，该类细胞的存活时长显著延长。在存活的Farage细胞中，与α干扰素（interferon-alpha）、同种异体移植排斥、核因子κB（NF-κB）、信号转导与转录激活因子3（STAT3）及信号转导与转录激活因子5（STAT5）相关的基因同样存在过表达现象。

总体实验设计：本研究设置了三组实验条件：
1）依据SLAMF1的表达水平对Farage细胞进行分选，分别标记为SLAM_high组与SLAM_low组；
2）将经1 μg/ml CHOP分别处理2天、3天的Farage细胞与未处理的Farage细胞进行对照；
3）将经1 μg/ml CHOP联合50 ng/ml IL4处理2天的Farage细胞与仅经50 ng/ml IL4处理的Farage细胞进行对照。

所有细胞系均采用Affymetrix HuGene ST 2.0芯片平台开展全基因组表达分析。差异表达基因采用微阵列统计分析（SAM）软件进行分析，并确立了特征基因集。