Introduction of functional chimeric E/L-selectin by RNA electroporation to target dendritic cells from blood to lymph nodes.
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.
Dörrie
Jan
J
Department of Dermatology, University Hospital Erlangen, Hartmannstr. 14, 91052 Erlangen, Germany.
Schaft
Niels
N
Müller
Ina
I
Wellner
Verena
V
Schunder
Tanja
T
Hänig
Jens
J
Oostingh
Gertie J
GJ
Schön
Michael P
MP
Robert
Caroline
C
Kämpgen
Eckhart
E
Schuler
Gerold
G
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
D019040:E-Selectin; D011993:Recombinant Fusion Proteins; D019041:L-Selectin; D012313:RNA