Microarray studies are increasingly used for toxicological research and even for the development of new toxicological test methods. Since gene-expression changes in cultured cells can be conveniently measured with microarrays, this method might be of use for in vitro toxicity testing, for example, in the field of contact sensitization. Allergic contact dermatitis, the clinical manifestation of contact sensitization, may occur when sensitizing chemicals enter the skin and get in contact with epidermal and dermal antigen-presenting cells.
View Article and Find Full Text PDFBackground: Animal tests have been used to characterize the potential of chemicals to produce allergic contact dermatitis, but this approach is increasingly a matter of public and political concern. Our aim was to develop and validate an alternative in vitro test that can identify contact allergens.
Methods: We developed a targeted microarray containing oligonucleotide probes for 66 immune-relevant genes and analyzed gene expression in monocyte-derived dendritic cells (Mo-DCs) treated with 1 irritant (SDS) and 2 prominent contact allergens, nickel and Bandrowski's base (BB), which is the oxidation product of the most important hair dye allergen, p-phenylenediamine.
Available ribonucleic acid (RNA) amplification methods are extensively tested for reproducibility, but only a few studies additionally deal with potential amplification bias. On targeted arrays, we evaluated three amplification protocols, which are less time consuming than the commonly used T7-RNA polymerase based in vitro transcription protocols and therefore may be more suitable for clinical use: Template-switching polymerase chain reaction (PCR), Ribo-single primer isothermal amplification and a random primer-based PCR. Additionally, a more sensitive labelling method, Dendrimer labelling, was evaluated.
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