Inhibitory effect of a modified adenovirus type 5E1A gene on the NF-κB activity in porcine aortic endothelial cells induced by TNF-α

The transcription factor nuclear factor κB (NF-κB) plays a key role in the delayed xenograft rejection (DXR). One of the important objects in the field is how to inhibit the NF-κB activity at optimal level. Thus, a modifiedE1A gene (E1AΔ) containing function domain (1–80 aa) and nuclear localization domain (139–243 aa) was used and cloned into an eucaryotic expression vector pcDNA3 to transfect the porcine aortic endothelial cells (PAEC). The stable transfectants were screened with G418.E1AΔ gene was able to be stably expressed in the PAEC and could not affect the growth of PAEC as analyzed by RT-PCR and cell growth rate. Reporter gene assay demonstrated thatE1AΔ was capable of inhibiting NF-κB activity in the PAEC induced by TNF-α without sensitizing to apoptosis, and the rate of inhibition was 53%. Furthermore,E1AΔ inhibited the expression of a NF-κB-dependent inflammatory geneE-selectin in the cells, and the rate of inhibition was 63%. In summary, the usage ofE1AΔ gene may be a new strategy to overcome DXR in the xenotransplantation.