Targeting gene delivery to activated vascular endothelium using anti E/P-Selectin antibody linked to PAMAM dendrimers.
BACKGROUND
The adhesion molecules P- and E-selectin are expressed on activated endothelial cells, and are good targets for gene therapy aimed at inflammatory disease. We have therefore investigated the potential of targeting PAMAM dendrimers, a non viral vector system, to cells expressing P/E-selectin using a monoclonal antibody that recognises these molecules.


METHODS
We used biotin and avidin to cross-link anti E/P-Selectin monoclonal antibody to pre-formed Superfect-DNA complexes that were then used to transfect reporter genes to CHO cells expressing E-Selectin, cytokine-activated primary Human Saphenous Vein Endothelial Cells (HSVEC) and whole vein segments.


RESULTS
The use of the anti E/P-Selectin antibody increased the transfection efficiency in CHO-E cells, activated HSVEC and saphenous vein segments ex vivo. We also showed that the antibody improved the binding of the complexes onto cells as well as the internalisation kinetics.


CONCLUSIONS
We demonstrate here that by attaching antibodies onto PAMAM dendrimers the efficiency of transfection can be significantly improved in cells or tissues expressing the receptor. This technology has potential in the treatment of cardiovascular disease by gene therapy but can also be used with different antibodies to target other diseased cells or tissues.
Theoharis
Stefanos
S
Immunology Department, Imperial College London, Hammersmith Hospital, Du Cane Rd, London W12 0NN, UK.
Krueger
Ute
U
Tan
Peng Hong
PH
Haskard
Dorian O
DO
Weber
Martin
M
George
Andrew J T
AJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D050091:Dendrimers / Q000378:metabolism*; D019040:E-Selectin / Q000276:immunology / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D015316:Genetic Therapy; D006801:Humans; D019007:P-Selectin / Q000276:immunology / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D000911:Antibodies, Monoclonal; D050091:Dendrimers; D019040:E-Selectin; D019007:P-Selectin