Specific stabilization of the 4F7 molecule on dendritic cells by contact allergens

Our laboratory has recently developed the monoclonal antibody 4F7 which recognizes a molecule on dendritic cells in the dermis of mice that is upregulated after application of contact allergens in vivo. Furthermore, this antibody detects an antigen on dendritic cells in spleen, lymph nodes and colon. In order to study the influence of contact allergens on the surface expression of the 4F7 molecules on dendritic cells, FACScan analysis of splenic dendritic cells was carried out after in vitro application of contact allergens. Freshly isolated splenic dendritic cells were found to be positive for 4F7, 33D1, N418 (CD11c) and MHC class II. After overnight culture the expression of the dendritic cell-specific molecules 4F7 and 33D1 was decreased. This downregulation was not inhibited by the addition of the cytokines TNF-alpha or GM-CSF during in vitro culture. However, in vitro treatment of freshly isolated dendritic cells with the contact allergen 2,4-dinitrofluorobenzene prevented this downregulation of the 4F7 surface molecules. The same effect was observed after treatment with other contact allergens (1-chloro-2,4-dinitrobenzene or potassium dichromate). Treatment with the irritant substance sodium dodecyl sulphate, the lectins concanavalin and lipopolysaccharide or the phorbol ester PMA did not prevent the downregulation of 4F7 and 33D1. Moreover, the influence of contact allergens on the expression of the molecules 4F7 and 33D1 was not inhibited by the protein synthesis inhibitor cycloheximide. No effects of contact sensitizers were detectable on the expression of MHC class II molecules or the costimulatory molecules B7 and heat-stable antigen. Our results show a specific stabilizing effect of contact allergens on the dendritic cell-specific molecules 4F7 and 33D1 independent of de novo protein synthesis.