In VivoTranstracheal Adenovirus-Mediated Transfer of Human Interleukin-10 Gene to Donor Lungs Ameliorates Ischemia-Reperfusion Injury and Improves Early Posttransplant Graft Function in the Rat

Abstract

We examined the effect of adenovirus-mediated transtracheal transfer of the human interleukin 10 (hIL-10) gene on lung ischemia-reperfusion (IR) injury, which is the insult due to hypothermic preservation plus graft reperfusion, and posttransplant lung function in Lewis rat lungs. Thirty rats were divided into 6 groups (n = 5). Groups 1 and 4 received 5 x 10(9) PFU of Ad5E1RSVhIL-10, groups 2 and 5 received 5 x 10(9) PFU of Ad5BGL2 ("empty" vector), and groups 3 and 6 received 3% sucrose (diluent). After 24 hr of in vivo transfection, lungs were stored at 4 degrees C (cold ischemic time, CIT) for 6 hr (groups 1-3) or 24 hr (groups 4-6) before transplantation. After 2 hr of reperfusion, lung function was assessed by oxygenation (FIO2, 1.0), airway pressure (AwP), and wet-to-dry (W/D) weight ratios. Rat tumor necrosis factor alpha (rTNF-alpha), interferon gamma (IFN-gamma), IL-10, and hIL-10 were measured in graft tissue and recipient plasma by ELISA and detected by immunohistochemistry (IHC). Partial pressure of oxygen (PaO2) levels in the hIL-10 group (6 hr of CIT) were higher than in empty vector and diluent groups (PaO2, 530 +/- 23 vs. 387 +/- 31 and 439 +/- 27 mmHg, respectively, p < 0.05). IL-10 rats after 24 hr of CIT showed higher PaO2 levels (260 +/- 29 mmHg) than empty vector (96 +/- 24 mmHg) or diluent (133 +/- 10 mmHg) lungs (p < 0.05). AwP and W/D ratios were reduced in hIL10 lungs (p < 0.05) compared with the other groups. rTNF-alpha and INF-gamma were reduced in tissue and plasma in groups 1 and 4 (p < 0.05). rIL-10 was reduced in the tissue of hIL-10 lungs (p < 0.05). IHC showed equal distribution of cytokines in tissue and abundant transgene expression in large and small airway epithelium in hIL-10 lungs.

Transbronchial administration of adenoviral-mediated interleukin-10 gene to the donor improves function in a pig lung transplant model

“…We have recently demonstrated that the transtracheal administration of adenoviralmediated human IL-10 to donor lungs 24 h before lung retrieval reduces ischemia-reperfusion injury and improves post-transplant graft function in a rat single lung transplant model. 16 Using the same rat model, we have also demonstrated that the transfection period could be safely reduced from 24 to 12 h, still achieving a similar improvement in lung function after transplantation in a more relevant timeframe clinically. 17 IL-10 is a potent anti-inflammatory cytokine that has a protective action in the mechanisms of ischemia-reperfusion injury.…”

“…16,17 The beneficial impact of IL-10 on ischemia-reperfusion injury has also been demonstrated by other investigators working in the field of lung transplantation. 18,19 However, these experiments were all performed in small animal models, and the applicability to large animals remains to be demonstrated before any potential application in humans appears feasible.…”

“…Hence, we performed a dose-escalation study to determine the minimal amount of vector to use to obtain significant expression of human IL-10 after 12 h of transfection. We progressively administered doses ranging from 5 Â 10 9 , which was effective in the rat, 16 Doses lower than 4 Â 10 10 pfu of AdhIL-10 did not provide measurable amounts of human IL-10 in the pig lung tissue. However, when transfection was carried out with 4 Â 10 10 pfu of AdhIL-10 administered to the left lung only (which is the lung used for transplantation in our model), a significant level of human IL-10 was detected in lung tissue after 12 h of transfection ( Figure 1).…”

“…Hence, as previously demonstrated in the rats, sufficient expression of human IL-10 before the period of cold ischemia has a beneficial effect on both the acute phase response to adenoviral vectors and ischemia-reperfusion-induced lung injury. 16,17 This finding highlights the importance of adequate expression of the transgene prior to lung retrieval in order to not only improve lung function, but also to avoid toxicity from IL-10 gene therapy for lung transplantation S Martins et al the adenoviral vector in the recipient -a critical issue if gene therapy is going to be applied clinically in lung transplantation. In the future, the development of more efficient and less inflammatory vectors such as gut-less adenovirus should limit this problem.…”

Recipient T Cells Mediate Reperfusion Injury after Lung Transplantation in the Rat

“…The peak airway pressures (PAwP) and arterial pressure of oxygen (PaO 2 ) of the transplanted lung have been shown to be reliable markers of lung function in a rat left single lung transplant model (23,25). PAwPs from the transplanted lung were measured with a three-way tap inserted between the endotracheal tube of the transplanted lung and the ventilator and connected to a pressure transducer.…”