Mapping of disease-associated expression polymorphisms in primary peripheral blood CD4+ lymphocytes. Homo sapiens

Analysis of expression quantitative trait loci (eQTLs) using RNA derived from freshly harvested peripheral blood CD4+ lymphocytes from 200 asthmatics collected in clinical settings. Overall design: RNA was obtained from peripheral blood CD4+ lymphocytes at four study centers. Peripheral blood (17 cc) was collected into BD Vacutainer CPT tubes (BD Diagnostics, Franklin Lakes, New Jersey) and placed on ice. Samples were centrifuged within 1 hour of collection for 20 minutes at 1700RCF, followed by mononuclear cell layer isolation and suspension in 10 ml of PBS. We isolated CD4+ lymphocytes using anti-CD4+ microbeads by column separation (Miltenyi Biotec, Auburn, CA) using 20 µl anti-CD4+ Micro beads per 106 total cells

本研究以临床收集的200名哮喘患者的新鲜采集外周血CD4+淋巴细胞提取的RNA为材料，开展表达数量性状基因座（expression quantitative trait loci, eQTLs）分析。实验设计概述：本研究在四个研究中心采集外周血CD4+淋巴细胞并提取RNA。具体样本处理流程如下：采集17毫升外周血至BD Vacutainer CPT采血管（BD诊断公司，新泽西州富兰克林莱克斯），并将样本置于冰上；样本于采集后1小时内以1700相对离心力离心20分钟，随后分离单个核细胞层并重悬于10毫升磷酸盐缓冲液（PBS）中。我们采用柱式分选法结合抗CD4+磁珠（Miltenyi Biotec公司，加利福尼亚州奥本）分离CD4+淋巴细胞，每10^6个总细胞使用20微升抗CD4+磁珠。