Clinical Chemistry
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Clinical Chemistry 52: 310-313, 2006; 10.1373/clinchem.2005.059774
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(Clinical Chemistry. 2006;52:310-313.)
© 2006 American Association for Clinical Chemistry, Inc.

Technical Briefs

Human ATP-Binding Cassette Transporter TaqMan Low-Density Array: Analysis of Macrophage Differentiation and Foam Cell Formation

Thomas Langmanna, Richard Mauerer and Gerd Schmitz

Institute of Clinical Chemistry, University of Regensburg, Regensburg, Germany;

aaddress correspondence to this author at: Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, D-93053 Regensburg, Germany; fax 49-941-944-6202, e-mail thomas.langmann{at}klinik.uni-regensburg.de


Abstract

Background: ATP-binding cassette (ABC) transporters cause various diseases and regulate many physiologic processes, such as lipid homeostasis, iron transport, and immune mechanisms. Several ABC transporters are involved in bile acid, phospholipid, and sterol transport, and their expression is itself controlled by lipids. In addition, ABC proteins mediate drug export in tumor cells and promote the development of multidrug resistance.

Methods: We created an ABC Transporter TaqMan Low-Density Array based on an Applied Biosystems 7900HT Micro Fluidic Card. We used a 2-µL reaction well with 2 ng of sample. To evaluate this method for lipidomic research and to characterize expression patterns of ABC transporters in cells relevant for atherosclerosis research, we monitored mRNA expression in human primary monocytes, in vitro–differentiated macrophages, and cells stimulated with the liver-X-receptor and retinoid-X-receptor agonists T0901317 and 9-cis retinoic acid, mimicking sterol loading.

Results: The method enabled simultaneous analysis of 47 human ABC transporters and the reference gene 18S rRNA in 2 replicates of 4 samples per run.

Conclusions: The new system uses only 2 ng of sample and small volumes of reagent, and the precaptured primers and probes avoided labor-intensive pipetting steps. The ABC Transporter TaqMan Low-Density Array may be a useful tool to monitor dysregulated ABC transporter mRNA profiles in human lipid disorders and cancer-related multidrug resistance and to analyze the pharmacologic and metabolic regulation of ABC transporter expression important for drug development in large-scale screening approaches.




The following articles in journals at HighWire Press have cited this article:


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 J. Leukoc. Biol.Home page
R. van de Ven, R. Oerlemans, J. W. van der Heijden, G. L. Scheffer, T. D. de Gruijl, G. Jansen, and R. J. Scheper
ABC drug transporters and immunity: novel therapeutic targets in autoimmunity and cancer
J. Leukoc. Biol., November 1, 2009; 86(5): 1075 - 1087.
[Abstract] [Full Text] [PDF]


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