Adantisense -ERK2 Gene Therapy Attenuates Chronic Rejection In Rat Aorta And Renal Transplantation

Objective: To construct recombinant adenoviral vector carring the anti-ERK2 gene of rat.Methods: The plasmid of p3XFLAG-CMV7.1-ERK2 was digested with enzymes, obtained fragment of ERK2cDNA,then connected it to T-vector. After sequencing, it was reversely inserted into the plasmid of pShuttle, at last transcribed it to AdEasy(tm) XL Adenoviral Vector System.Results: The plasmid of pshuttle-anti-ERK2 was digested with DraI and NotI enzymes, and found that the sequence and transcription orientation were identical with expectation.Conclusion: Recombinant adenoviral vector carring the anti-ERK2 gene been constructed successfully by reverse orientation cloning the target fragment into pshuttle. It provided tools for researching the role of anti-ERK2 gene therapy in graft rejection. Objective: To investigate the protective effects on aorta and renal allografts and the possible mechanism of Adantisense–ERK2 gene therapy against chronic rejection injury.Methods: Ex vivo Adantisense-ERK2 gene therapy was performed in vascular and renal transplantation models. At 60 days after transplantation in aorta transplantation and at 90 days in renal model, we analysed the structure and function of allografts, the expression of therapeutic genes and proteins, and the immune modulation.Results: Adantisense-ERK2 gene therapy reduces ERK protein expression in allografts and protected allogrfts against chronic rejection injury. The transfected empty vehicle aggravated chronic rejection damage in aortra and renal allografts. Adantisense–ERK2 gene therapy decreased the infiltration of macrophages and T lymphocyte cells, and also reduced TNF-αand ICAM-1 expression in serum.Conclusion: Adanti-ERK2 gene therapy can protect aorta and renal allografts against chronic rejection injury, the protective role may correlate with decreased TNF-αand ICAM-1expression, and inhibition the infiltration of immune cells.