title: Allergen-induced changes in interleukin 1 beta (IL-1 beta) mRNA expression by human blood-derived dendritic cells: inter-individual differences and relevance for sensitization testing.
abstract: The development of in vitro methods for the identification of skin sensitizers based upon analysis of Langerhans cell (LC) function has been constrained by the fact that these cells represent only a minority population in the skin that, once isolated, alter their phenotype spontaneously and rapidly. Methods have been developed recently that allow the expansion in culture using appropriate cytokine conditions of LC-like dendritic cells (DCs) from certain tissues, including human peripheral blood. It has been demonstrated that culture of human blood-derived LC-like cells with selected potent contact allergens such as 2,4-dinitrofluorobenzene (DNFB) stimulates selective phenotypic changes, including the up-regulation of interleukin 1 beta (IL-1 beta) mRNA expression, under conditions where skin irritants are without effect. However, in our own previous investigations, we have observed that there appear to be differences between blood donors with respect to the responsiveness of DCs to DNFB-induced changes in IL-1 beta expression, differences that could compromise the utility of this approach as a screening method for contact allergens. We have therefore investigated donor variability in DC responsiveness to a panel of known human contact allergens (DNFB; paraphenylene diamine, PPD; methyl- chloroisothiazolinone/methylisothiazolinone, CMIT), to the skin irritant benzalkonium chloride and to the mitogen phorbol myristate acetate (PMA). Dendritic cells derived from all donors expressed IL-1 beta mRNA constitutively. Treatment of DCs isolated from donors with a responder phenotype to DNFB with PPD or CMIT resulted also in up-regulation of IL-1 beta mRNA expression, although such changes were always comparatively modest, generally resulting in a twofold induction compared with vehicle-treated controls. Dendritic cells derived from donors with a non-responder phenotype to DNFB failed also to respond to these additional contact allergens under conditions where the mitogen PMA caused similar increases in IL-1 beta expression to those observed for allergen-responsive donors. Benzalkonium chloride failed to provoke changes in the expression of this cytokine in any donor examined, irrespective of their responder phenotype. The temporal stability of the responder/non-responder DC phenotype was confirmed, with stable phenotypes with respect to DNFB-induced changes in IL-1 beta mRNA expression observed over a period of some 18 months. Fifty per cent (6/12) of donors tested over this period displayed a responder phenotype. These data demonstrate that chemical allergens do stimulate consistent changes in IL-1 beta mRNA expression in the proportion of donors who have a responsive phenotype, and that such responses are apparently selective for allergen using the relatively narrow range of materials assessed to date. However, the modest response to very strong contact allergens, coupled with the difficulties of responder/non-responder phenotypes, means that in its present form this approach does not lend itself to the routine assessment of skin sensitizing activity.
authors: Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire SK10 4TJ, UK.
authors: J S
authors: JS
authors: Pichowski
authors: M
authors: M
authors: Cumberbatch
authors: R J
authors: RJ
authors: Dearman
authors: D A
authors: DA
authors: Basketter
authors: I
authors: I
authors: Kimber
mesh_terms: D000328:Adult; D000485:Allergens / Q000276:immunology*; D001681:Biological Assay / Q000379:methods; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology; D003877:Dermatitis, Contact / Q000276:immunology*; D005260:Female; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis*; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D012641:Selection, Genetic
chemical_list: D000485:Allergens; D007375:Interleukin-1; D012333:RNA, Messenger