Gene-expression monitoring in dendritic cells after stimulation with potential allergens - DNA microarrays and real-time PCR opening new strategies in toxicological testing

  • Nöhammer, C. (Author)
  • Waltraud Novak (Author)
  • Roland Schafleitner (Author)
  • Helga Tuschl (Author)

Activity: Talk or presentation / LecturePresentation at a scientific conference / workshop

Description

Motivation and background for the present research was the development of an in vitro toxicological test system for reliable evaluation and prediction of the sensitising potential of chemicals. To this end gene expression of 20 different cytokines was analysed in human dendritic cells after stimulation with contact allergens (NiSO4, cinnamaldehyd, Bandrowski base) or irritants (SDS). For identification of potential stressing agent-specific changes in gene expression we established real-time PCRs for each of the 20 investigated cytokine and chemokine genes as well as for 7 house keeping genes. In parallel we designed gene-specific 50-mer oligonucleotide probes to be included in a DNA microarray, which was finally optimized to give specific signals using only 2 g of total RNA. Both methods consistently exhibited varying expression profiles depending on the stressing compound. In detail our studies revealed that cultured dendritic cells stressed with nickel show strong transcriptional activation of IL-1#alpha, IL-1ß, MIP-1#alpha, and MIP-2#alpha whereas Bandrowski base treated cells exhibit only slightly increased expression of IL-1ß, MIP-1#alpha, MIP-2#alpha and TNF-#alpha. Cinnamaldehyde stimulation led to a prominent induction of IL-1#alpha and MIP-2#alpha. In contrast to NiSO4, cinnamaldehyd and Bandrowski base, all representing potential allergens, the irritant SDS provoked only very modest changes in dendritic cell cytokine expression. After stimulation with SDS the extent of induction pretty much varied depending on the individual donor of dendritic cells and was only consistent for IL-6 and IL12p35. From our preliminary results we conclude that the analysis of cytokine expression profiles allows to predict the allergenicity of chemicals and to classify them as sensitisers.
Period26 Jan 200427 Jan 2004
Event titleStatusseminar: Chiptechnologien / Microarrays, Hygiene, Gesundheit
Event typeOther
Degree of RecognitionInternational

Research Field

  • Not defined