title: Cellular handling of a dexamethasone-anti-E-selectin immunoconjugate by activated endothelial cells: comparison with free dexamethasone.
abstract: OBJECTIVE
  For selective inhibition of endothelial cell activation in chronic inflammation, we have developed a dexamethasone-anti-E-selectin immunoconjugate. The present study was performed to evaluate the cellular handling of this immunoconjugate by activated primary endothelial cells and to compare its drug delivery capacity with free dexamethasone.
  
  
  METHODS
  The binding, uptake, and degradation of 125I-radiolabeled dexamethasone-anti-E-selectin immunoconjugate by TNFalpha-activated endothelial cells were studied for different time periods and at different concentrations, as well as in the presence of inhibitors for E-selectin binding and lysosomal degradation. Its drug delivery capacity was compared with the uptake of unconjugated 3H-labeled dexamethasone.
  
  
  RESULTS
  The immunoconjugate was internalized by E-selectin expressing activated endothelial cells and degraded in the lysosomal compartment. The receptor-mediated binding and uptake was saturable, implying a maximal attainable intracellular concentration of the drug. In contrast, free dexamethasone entered both resting and activated endothelial cells by passive diffusion.
  
  
  CONCLUSIONS
  The dexamethasone-anti-E-selectin immunoconjugate is capable of selective delivering the coupled drug into activated endothelial cells. This targeting concept enables disease-induced drug delivery in which intracellular concentrations can be reached comparable with those obtained after incubation with 3 FM dexamethasone.
authors.affiliation: Department of Pharmacokinetics and Drug Delivery, University Centre for Pharmacy, Groningen University Institute for Drug Exploration (GUIDE), Groningen, The Netherlands. r.j.kok@farm.rug.nl
authors.forename: Robbert J
authors.initials: RJ
authors.lastname: Kok
authors.forename: Maaike
authors.initials: M
authors.lastname: Everts
authors.forename: Sigridur A
authors.initials: SA
authors.lastname: Asgeirsdóttir
authors.forename: Dirk K F
authors.initials: DK
authors.lastname: Meijer
authors.forename: Grietje
authors.initials: G
authors.lastname: Molema
mesh_terms: D002478:Cells, Cultured; D002626:Chemistry, Pharmaceutical; D003907:Dexamethasone / Q000493:pharmacokinetics*; D019040:E-Selectin / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D018796:Immunoconjugates / Q000493:pharmacokinetics*
chemical_list: D019040:E-Selectin; D018796:Immunoconjugates; D003907:Dexamethasone