Gene expression profiling in human peripheral blood mononuclear cells using high-density filter-based cDNA microarrays

被引:36
作者
Walker, J [1 ]
Rigley, K [1 ]
机构
[1] Edward Jenner Inst Vaccine Res, Dendritic Cell Grp, Newbury RG20 7NN, Berks, England
关键词
microarray; gene expression; cluster analysis; PBMCs;
D O I
10.1016/S0022-1759(00)00181-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Microarray technology has provided the ability to analyse the expression profiles for thousands of genes in parallel. The need for highly specialised equipment to use certain types of microarrays has restricted the application of this technology to a small number of dedicated laboratories. High-density filter-based cDNA microarrays provide a low-cost option for performing high-throughput gene expression analysis. We have used a model system in which filter-based cDNA microarrays representing over 4000 known human genes were used to monitor the kinetics of gene expression in human peripheral blood mononuclear cells (PBMCs) stimulated with phytohaemagluttinin (PHA). Using software-based cluster analysis, we identified 104 genes that altered in expression levels in response to PHA stimulation of PBMCs and showed that there was a considerable overlap between gents with similar temporal expression profiles and similar functional roles. Comparison of microarray quantitation with quantitative PCR showed almost identical expression profiles for a number of genes. Coupled with the fact that our findings are in agreement with a large number of independent observations, we conclude that the use of filter-based cDNA microarrays is a valid and accurate method for high-throughput gene expression profiling. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:167 / 179
页数:13
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