Effect of adenoviral titer and instillation pressure on gene transfer efficiency to arterial and venous grafts ex-vivo.
Effect of adenoviral titer and instillation pressure on gene transfer efficiency to arterial and venous grafts ex-vivo.
复制标题
腺病毒滴度和滴注压力对离体动脉和静脉移植物基因转移效率的影响。
DOI：
10.1067/mva.2002.125029
复制
发表时间：
2002
影响因子：
4.3
通讯作者：
Messina,LouisM
中科院分区：
文献类型：
--
作者：
Brevetti,LucyS;Chang,DavidS;Sarkar,Rajabrata;Messina,LouisM
ObjectiveAdenoviral-mediated gene transfer to arterial and venous grafts has potential in the treatment of a number of vascular diseases. Despite widespread use of these vectors to mediate gene transfer to blood vessel walls, the optimal transduction conditions for each type of vessel has yet to be determined. Our objective was to study the effect of adenoviral titer and instillation pressure on efficiency of gene transfer to arterial and venous grafts ex-vivo.MethodsJugular vein and carotid artery segments of 8 cm were harvested from Yorkshire Cross pigs. Tissue culture media or different titers of an adenoviral vector encoding human placental alkaline phosphatase (hpAP) were instilled into venous and arterial grafts at 0 mm Hg or 80 to 100 mm Hg of pressure and bathed externally in the same solution at 37° C for 30 minutes. The grafts were rinsed, opened longitudinally, and incubated in culture media at 37° C for 48 hours. Grafts were fixed and stained for hpAP transgene expression to quantitate percent luminal transduction or homogenized for alkaline phosphatase (AP) activity to determine total transmural transduction.ResultsFor venous grafts, the percent luminal area stained for hpAP was greatest with 108plaque-forming units/mL at 0 mm Hg (81% ± 7%) and decreased with increasing titers (53% ± 9% at 109pfu/mL and 44% ± 11% at 5 × 109pfu/mL; n = 7; P < .05). No increase in percent luminal area stain was achieved with an instillation pressure of 80 to 100 mm Hg at any viral titer. The inverse finding was observed in arterial grafts. For arterial grafts, the greatest percent luminal area stained was achieved with 5 × 109pfu/mL at 80 to 100 mm Hg (76% ± 7%). An instillation pressure of 80 to 100 mm Hg increased the percent luminal area stained at 108pfu/mL from 31% ± 9% to 66% ± 8% (n = 8; P = .01). For venous grafts, total AP activity peaked with 109pfu/mL at 0 mm Hg and decreased with an instillation pressure of 80 to 100 mm Hg (30.6 ± 9.7 U/mg versus 10.9 ± 2.5 U/mg; n = 7; P < .01). However, for arterial grafts, total AP activity peaked with 5 × 109pfu/mL (0 mm Hg) and increased with an instillation pressure of 80 to 100 mm Hg (32.8 ± 9.9 U/mg versus 63.4 ± 20.5 U/mg; n = 8; P < .05).ConclusionHigh transduction efficiency can be achieved with adenoviral-mediated gene transfer of arterial and venous grafts. Gene transfer with the vascular graft's physiologic pressure conditions improved transduction efficiency for the artery (80 to 100 mm Hg) and vein (0 mm Hg). Comprehensive analysis of adenoviral transduction conditions is important to realize the full promise of adenoviral-mediated gene transfer. (J Vasc Surg 2002;36:263-70.)
