Journal Article

Loss of FADS2 Function Severely Impairs the Use of HeLa Cells as an In Vitro Model for Host Response Studies Involving Fatty Acid Effects

Schlörmann, Wiebke, Rohrer, Carsten, Jaudszus, Anke, Degen, Christian, Jahreis, Gerhard, Sauerwein, Helga, Sachse, Konrad, Barth, Stephan W., Klempt, Martin, Roth, Alexander

2014

Overview

Established epithelial cell lines equipped with pattern recognition receptors such as the Toll-like receptor (TLR)-2 are common tools for immune response studies on invading pathogens, e.g. the obligate intracellular species of Chlamydia. Moreover, such models are widely used to elucidate fatty acid-mediated immune effects. In several transformed cell lines, however, unusual loss of metabolic functions was described. The cell lines A549 and HeLa are poorly characterized in this respect. Therefore, we comparatively assessed the metabolic capacity of A549 and HeLa prior to proposed application as in vitro model for fatty acid effects on chlamydial infection. We incubated both cell lines either with substrates (C18:2n-6 or C18:3n-3) or products (C18:3n-6, C18:4n-3) of fatty acid desaturase-2 (FADS2), and analysed the fatty acid profiles after 24 h and 72 h by gas chromatography. Based on these data, we suspected that the complete discontinuation of normal biosynthesis of long-chain polyunsaturated fatty acids (LC-PUFA) in HeLa was due to loss of FADS2 function. Consequently, prostaglandin E2 (PGE2) formation was less inducible by TLR2 stimulation in HeLa, likely as a result of not only insufficient supply of precursors but also weak cyclooxygenase-2 (COX-2) response. In accordance, Chlamydia infection rates were consistently lower in HeLa than in A549. Sequence analysis revealed no alteration within the FADS2 gene in HeLa. The FADS2 expression level, however, was significantly lower and, in contrast to A549, not regulated by C18:2n-6. A549 exhibited regular fatty acid metabolism and enzyme functionality. Our data show that HeLa cells considerably differ from A549 at several stages of fatty acid metabolism. The poor metabolic potential of HeLa, mainly concerning FADS2 upstream of COX-2 function, calls into question whether these cells represent a good model to unveil fatty acid or downstream eicosanoid effects in the course of intracellular bacterial infection.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Bacteria / Bacterial diseases / Biochemistry / Biology and life sciences / Biosynthesis / Biotechnology / Cell lines / Chlamydia / Chlamydia - metabolism / Chlamydia Infections - genetics / Chlamydia Infections - metabolism / Chlamydiaceae / Cyclooxygenase 2 - genetics / Cyclooxygenase 2 - metabolism / Cyclooxygenase-2 / Data processing / Desaturase / Dinoprostone - genetics / Dinoprostone - metabolism / Epithelial cells / Fatty Acid Desaturases - deficiency / Fatty acids / Fatty Acids - genetics / Fatty Acids - metabolism / Flow cytometry / Food / Gas chromatography / Gene expression / HeLa Cells / Humans / Immune response / Immune system / Infections / Intracellular / Kinases / Lipids / Metabolism / Nutrition / Pattern recognition / Pattern recognition receptors / Physiology / Polyunsaturated fatty acids / Prostaglandin E2 / Proteins / Receptors / Rodents / Sexually transmitted diseases / STD / Substrates / TLR2 protein / Toll-Like Receptor 2 - genetics / Toll-Like Receptor 2 - metabolism / Toll-like receptors