title: Gene expression changes in peripheral blood-derived dendritic cells following exposure to a contact allergen.
abstract: A critical step in the induction of allergic contact allergy is the activation and subsequent migration of Langerhans cells (LC), an important antigen presenting dendritic cell (DC) of the skin. As the Langerhans cells migrate, they undergo a maturation process. It has been proposed that contact allergen exposure can induce DC maturation. While changes in DC gene expression profiles induced by various maturation stimuli have been explored, there are no published reports describing genomic-scale analysis of the changes induced by chemical allergen exposure. Therefore, to explore the concept of chemical allergen-induced DC maturation and to identify genes that are regulated by exposure to allergens we examined, at the transcriptional level, the effects of exposure to a contact allergen on DC. Peripheral blood-derived DC were exposed for 24 h to either 1mM or 5 mM dinitrobenzenesulfonic acid (DNBS). Changes in gene expression were analyzed using Affymetrix U95Av2 GeneChip. Comparison of mean signal values from replicate cultures revealed 173 genes that were significantly different (P < or = 0.001) between 1 mM DNBS treated and untreated control DC and 1249 significant gene changes between 5 mM DNBS treated and control DC. Real-time reverse-transcriptase polymerase chain reaction (RT-PCR) was used to evaluate the observed transcript changes for selected genes in DC derived from a second donor. Comparison of the fold-changes in transcript levels between the two platforms and donors revealed a good correlation in both direction and magnitude. RT-PCR analysis was also used to assess the allergen specificity of a selected number of genes in DC derived from a third donor. Many of the gene expression changes were found to be induced only by exposure to the allergen, DNBS, and not by exposure to a structurally similar non-allergen, benzenesulfonic acid. A number of gene expression changes induced by allergen exposure were found to be consistent with what is known of the DC maturation process, and thus provide support for the theory of contact allergen-induced DC maturation. Additionally, it is hoped that some of the transcript changes identified through this approach will be shown to be suitable for use in the development of an in vitro predictive assay for contact sensitization.
authors.affiliation: Miami Valley Laboratories, Central Product Safety Department, The Procter & Gamble Company, P.O. Box 538707, Cincinnati, OH 45253, USA. ryan.ca@pg.com
authors.forename: Cindy A
authors.initials: CA
authors.lastname: Ryan
authors.forename: Lucy A
authors.initials: LA
authors.lastname: Gildea
authors.forename: Ben C
authors.initials: BC
authors.lastname: Hulette
authors.forename: Rebecca J
authors.initials: RJ
authors.lastname: Dearman
authors.forename: Ian
authors.initials: I
authors.lastname: Kimber
authors.forename: G Frank
authors.initials: GF
authors.lastname: Gerberick
mesh_terms: D000485:Allergens / Q000494:pharmacology*; D001557:Benzenesulfonates / Q000494:pharmacology*; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000187:drug effects* / Q000378:metabolism; D017449:Dermatitis, Allergic Contact / Q000209:etiology*; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000485:Allergens; D001557:Benzenesulfonates; C007488:dinitrobenzenesulfonic acid