Adenovirus-mediated gene transfer of triple human complement regulating proteins (DAF, MCP and CD59) in the xenogeneic porcine-to-human transplantation model. Part II: xenogeneic perfusion of the porcine liver in vivo

Abstract

In this study, the adenovirus-mediated gene transfer of triple human complement regulating proteins was investigated in xenogeneic pig liver perfusion. The porcine liver was perfused in situ at 4 "C under a pump-driven veno-venous shunt of the portal vein and inferior vena cava, with 5 to 1 5~1 0~' plaqueforming units (pfu) of adenovirus vector (group 1: AxCALacZ; 2: Ax-CADAF; 4: AxCACD59 + AxCADAF + AxCAMCP) for 1 h (for each, n=3). The livers were harvested 24 h after gene transfer and then were reperfused ex-vivo with fresh human blood for 2 h. In immunohistochemical staining, each complement regulating protein (CRP) showed a distribution similar to that of the LacZ expression. The CACD59; 3: AxCACD59 + AX-C3 levels in the perfusate were also maintained at higher levels in group 4 from 60 to 120 min after reperfusion (C3: 85% to 95% of the initial level) than in groups 1 to 3 (C3: 80% to 90% of the initial level) from 60 to 120 min after reperfusion. The complement deposition on the porcine liver [C3, membrane attack component (MAC)] decreased significantly more in group 4 than in groups 1 to 3. In conclusion, the adenovirus-mediated multiple gene transfer of human CRPs (hCRPs) was found to effectively suppress the complement activation in xenogeneic pig liver perfusion.

Transgenic pigs expressing human CD59, in combination with human membrane cofactor protein and human decay‐accelerating factor

“…It was hypothesized that expression of hCD59 in combination with hMCP and hDAF could provide more effective protection to the transplanted pig tissues. Shiraishi et al [9] reported that complement deposition was decreased significantly in the xenogeneic porcine liver perfusion model after adenovirus‐mediated gene transfer of DAF, MCP, and CD59. In the present study, we report the production of pigs transgenic for hCD59 alone or in combination with hMCP and hDAF, and the initial investigation of the ability of these transgenes expressed on pig cells to regulate human complement activation.…”