Expression of the chemokine antagonist vMIP II using a non-viral vector can prolong corneal allograft survival.
BACKGROUND
The expression of chemokines is central to the recruitment of inflammatory cells for graft rejection, and modulation of chemokine action is of potential in preventing graft rejection. We have examined chemokine expression in a murine model of corneal allograft rejection, and also determined the effect of expressing a broad acting chemokine antagonist, viral macrophage inflammatory protein II (vMIP II), on graft survival.


METHODS
The expression of chemokines in a murine model of corneal transplantation was determined by real time RT-PCR and, in the case of regulated on activation normal T-cell expressed and secreted, by ELISA. The plasmid encoding the virally derived chemokine antagonist, vMIP II, was introduced into the corneal endothelial cells using a non-viral vector consisting of liposomes and transferrin. The expression and activity of vMIP II was determined by ELISA and functional assays, and the effect on graft survival noted.


RESULTS
After allotransplantation, there was up-regulation of all 11 chemokines examined. After gene delivery, there was expression of active vMIP II for more than 14 days and considerable prolongation of graft survival. This was associated with a decrease in leukocyte infiltration of the stroma of the cells.


CONCLUSIONS
As expected there was considerable up-regulation of chemokines during allograft rejection. The expression of vMIP II showed considerable prolongation of graft survival. This is the first time we have observed prolongation of graft survival after a non-viral (as opposed to viral) means of gene delivery and indicates the potential of interfering with chemokine action to prevent corneal graft failure.
Pillai
Radhakrishna G
RG
Department of Immunology, Division of Medicine, Imperial College London, Hammersmith Campus, London, UK.
Beutelspacher
Sven C
SC
Larkin
D Frank P
DF
George
Andrew J T
AJ
D000818:Animals; D015153:Blotting, Western; D018925:Chemokines / Q000235:genetics*; D019742:Chemokines, CC; D016039:Corneal Transplantation / Q000502:physiology*; D004195:Disease Models, Animal; D005500:Follow-Up Studies; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage*; D006085:Graft Survival / Q000235:genetics*; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D011379:Prognosis; D012313:RNA / Q000235:genetics*; D019707:Receptors, Chemokine / Q000037:antagonists & inhibitors*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014184:Transplantation, Homologous
D018925:Chemokines; D019742:Chemokines, CC; D019707:Receptors, Chemokine; C108059:vMIP-II; D012313:RNA