Contact sensitizers modulate mechanisms of receptor-mediated endocytosis but not fluid-phase endocytosis in murine epidermal Langerhans cells.
In order to define the influence of contact allergens on the fluid-phase endocytosis (FPE) of soluble molecules of murine epidermal Langerhans cells (LC), we studied the internalization of FITC-labeled bovine serum albumin (FITC-BSA), TRITC-labeled dextrane (TRITC-DEX) as well as horseradish peroxidase by LC. A 3-parameter flow-cytometric technique was performed for quantification of internalized FITC-BSA in LC using quantum red-labeled reagents for detection of Ia-antigen expression by LC and propidium iodide for exclusion of dead cells from analysis. A temperature-dependent rapid accumulation of FITC-BSA was noticed in time-course studies reaching a plateau between 1 and 2 h of in vitro culture at 37 degrees C. The quantity of FPE under stimulation with phorbol 12-myristate 13-acetate (PMA), concanavalin A (Con A), staphylococcal enterotoxin B (SEB) and contact sensitizers (DNFB, Kathon CG, K2Cr2O7) as well as the irritant SLS was determined. Treatment of LC with PMA and Con A resulted in a significant increase of total FITC-BSA uptake. The contact sensitizers as well as SEB and SLS failed to mediate augmented fluid-phase endocytosis. By use of the pH-insensitive soluble marker, TRITC-DEX and a microscope photometer for evaluation these findings could be confirmed. This excluded any artificial influence of differences in pH values in endocytotic compartments which might have influenced the fluorescence intensity of the pH-sensitive fluorochrome FITC. For qualitative analysis of FPE, the intracellular distribution of internalized horseradish peroxidase in LC was studied. An aggregated pattern became apparent in untreated LC and did not change under stimulation with any of the substances used.(ABSTRACT TRUNCATED AT 250 WORDS)
Becker
D
D
Hautklinik der Johannes Gutenberg-Universität, Mainz, Germany.
Lempertz
U
U
Enk
A
A
Saloga
J
J
Knop
J
J
D000818:Animals; D002273:Carcinogens / Q000494:pharmacology; D003208:Concanavalin A / Q000494:pharmacology; D017449:Dermatitis, Allergic Contact / Q000378:metabolism / Q000473:pathology*; D004139:Dinitrofluorobenzene / Q000494:pharmacology; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D007801:Langerhans Cells / Q000187:drug effects*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010873:Pinocytosis / Q000187:drug effects*; D012235:Rhodamines / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D002273:Carcinogens; D012235:Rhodamines; C039217:fluorescein isothiocyanate bovine serum albumin; D003208:Concanavalin A; D012710:Serum Albumin, Bovine; C009434:tetramethylrhodamine isothiocyanate; D004139:Dinitrofluorobenzene; D016650:Fluorescein-5-isothiocyanate; D013755:Tetradecanoylphorbol Acetate