Effects of vaccinia virus anti-inflammatory protein 35K and TIMP-1 gene transfers on vein graft stenosis in rabbits.
BACKGROUND
Vein graft stenosis is a common problem after bypass surgery. Vein grafts are ideal targets for gene therapy because transduction can be made ex vivo before grafting. Since chemokines and inflammatory factors are involved in vein graft thickening, we tested a hypothesis that the vaccinia virus anti-inflammatory protein 35K which can sequester CC-chemokines, can reduce vein graft thickening in vivo.


METHODS
We used adenovirus-mediated gene transfer (1x10(9) pfu/ml) of 35K and compared its effects on reducing stenosis in a rabbit jugular vein graft model with tissue inhibitor of metalloproteinase-1 (TIMP-1) and LacZ control gene. TIMP-1 was used in this study because it has previously been shown to inhibit vein graft stenosis in other model systems. The expression of transgenes in the transduced segments was confirmed by RT-PCR. Vein grafts were analyzed using immunohistological and morphometric methods at the three-day time-point and at two-week and four-week time-points.


RESULTS
It was found that the anti-inflammatory protein 35K was an efficient factor in reducing neointima formation at the two-week time-point, indicating that inflammatory factors play an important role in vein graft stenosis. At the four-week time-point, 35K still showed a reduced accumulation of macrophages. TIMP-1 also tended to reduce neointimal thickening at the two-week time-point as compared to LacZ.


CONCLUSIONS
It was found that 35K is an efficient factor in reducing neointima formation, macrophage accumulation and proliferation in rabbit vein grafts after adenoviral ex vivo gene transfer.
Puhakka
Hanna L
HL
A. I. Virtanen Institute, University of Kuopio, Kuopio, Finland.
Turunen
Päivi
P
Gruchala
Marcin
M
Bursill
Christine
C
Heikura
Tommi
T
Vajanto
Ismo
I
Greaves
David R
DR
Channons
Keith
K
Ylä-Herttuala
Seppo
S
D000256:Adenoviridae / Q000235:genetics*; D000714:Anastomosis, Surgical; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D017536:Carotid Artery, Common / Q000601:surgery; D002460:Cell Line; D003251:Constriction, Pathologic / Q000517:prevention & control*; D017931:DNA Primers; D018014:Gene Transfer Techniques; D006801:Humans; D007601:Jugular Veins / Q000187:drug effects / Q000502:physiology / Q000601:surgery; D008297:Male; D018919:Neovascularization, Physiologic; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics* / Q000494:pharmacology; D017539:Tunica Intima / Q000166:cytology; D020246:Venous Thrombosis / Q000628:therapy*; D014759:Viral Envelope Proteins / Q000494:pharmacology*; D014764:Viral Proteins / Q000494:pharmacology*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D017931:DNA Primers; D019715:Tissue Inhibitor of Metalloproteinase-1; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C072495:immunodominant protein p35, vaccinia virus