Guironnet, G., C. Dalbiez-Gauthier, F. Rousset, D. Schmitt and J. Peguet-Navarro. (2000).In Vitro Human T Cell Sensitization to Haptens by Monocyte-Derived Dendritic Cells Toxicology in Vitro 14: 517-522. [Reprinted with permission from Elsevier Science]

fluorescein isothiocyanate - 27072-45-3; eugenol - 00097-53-0; isoeugenol - 00097-54-1

We previously reported that in vitro primary sensitization of hapten-specific T cells by cultured human epidermal Langerhans cells (LC) provides an alternative approach to discriminate strong contact sensitizers from irritants (Krasteva et al, 1996; Moulon et al., 1993). However, this LC-based immunoassay was limited by the availability of human skin samples. In the present study, we used monocyte-derived dendritic cells (DC) to analyse the autologous proliferative T cell response to several allergens. Monocytes were purified from the peripheral blood of healthy donors and cultured for 6-8 days in the presence of GM/CSF and IL-4 and then for 2 days in the presence of GM/CSF and TNF-alpha. The resulting cells exhibited the phenotype of mature DC, as assessed by the strong expression of HLA-DR, CD80, CD83 and CD86 antigens. We showed that trinitrophenyl (TNP) treated mature DC induced a significant T cell proliferative response in all experiments, while fluorescein isothiocyanate (FITC) gave positive results in about half of them. The prohaptens eugenol and isoeugenol induced significant proliferation in one out of eight and in four out of 12 experiments, respectively. Interestingly, in 16 assays T cells never proliferated in the presence of sodium lauryl sulfate (SLS)-treated DC. Thus, this in vitro model allows discrimination between strong contact sensitizers and irritants. It might be very useful, therefore, for restriction of animal experimentation.