title: Introduction of functional chimeric E/L-selectin by RNA electroporation to target dendritic cells from blood to lymph nodes.
abstract: BACKGROUND
  Inefficient migration of dendritic cells (DC) to regional lymph nodes (LN) upon intracutaneous injection is a major obstacle for effective DC vaccination. Intravenous vaccination is unfavorable, because DC cannot migrate directly from the blood into LN.
  
  
  METHODS
  To enable human monocyte-derived (mo)DC to enter LN directly from the blood, we manipulated them by RNA electroporation to express a human chimeric E/L-selectin (CD62E/CD62L) protein, which binds to peripheral node addressin expressed on high endothelial venules.
  
  
  RESULTS
  Transfection efficiency exceeded 95%, and high E/L-selectin surface expression was detected for >48 h. E/L-selectin RNA-transfected DC displayed an identical mature DC phenotype as mock-transfected DC. Furthermore, E/L-selectin-transfected DC maintained their normal CCR7-mediated migration capacity, and their ability to prime and expand functional cytotoxic T cells recognizing MelanA. Most importantly, E/L-selectin-RNA-transfected DC gained the capability to attach to and roll on sialyl-Lewis(X) in vitro.
  
  
  CONCLUSIONS
  The presented strategy can be readily translated into the clinic, as it involves no stable genetic manipulation or viral transformation, and allows targeting of a large number of LN.
authors.affiliation: Department of Dermatology, University Hospital Erlangen, Hartmannstr. 14, 91052 Erlangen, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Dörrie
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
authors.forename: Ina
authors.initials: I
authors.lastname: Müller
authors.forename: Verena
authors.initials: V
authors.lastname: Wellner
authors.forename: Tanja
authors.initials: T
authors.lastname: Schunder
authors.forename: Jens
authors.initials: J
authors.lastname: Hänig
authors.forename: Gertie J
authors.initials: GJ
authors.lastname: Oostingh
authors.forename: Michael P
authors.initials: MP
authors.lastname: Schön
authors.forename: Caroline
authors.initials: C
authors.lastname: Robert
authors.forename: Eckhart
authors.initials: E
authors.lastname: Kämpgen
authors.forename: Gerold
authors.initials: G
authors.lastname: Schuler
mesh_terms: D002465:Cell Movement / Q000502:physiology*; D002678:Chimera; D003713:Dendritic Cells / Q000378:metabolism*; D019040:E-Selectin / Q000378:metabolism*; D018274:Electroporation; D005434:Flow Cytometry; D006801:Humans; D019041:L-Selectin / Q000378:metabolism*; D008198:Lymph Nodes / Q000166:cytology / Q000378:metabolism; D012313:RNA; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection; D014611:Vaccination / Q000379:methods
chemical_list: D019040:E-Selectin; D011993:Recombinant Fusion Proteins; D019041:L-Selectin; D012313:RNA