人表面抗体基因在肝移植中抑制乙型肝炎病毒再感染

To investigate gene transfer of denovirus vector for inducing immune tolerance of liver graft by blockade of T cell costimulatory pathway, we constructed an adenovirus shuttle plasmid containing human CTLA4-Ig recombinant fusion gene and cotransfected with an intact supercoiled Ad backbone plasmid named pAdEasy-1/△E1、△E 3 into E. Coli strain BJ5183 for preparing recombinant adenovirus plasmid pAd-Tracker-CTLA4-Ig. Subsequently, it was transfected into 293 cells for packaging recombinant adenovirus d-Tracker-CTLA4-Ig. After infecting LO2 cells with Ad-Tracker-CTLA4-Ig for 72 hours, the expression of protein CTLA4-Ig was detected by ELISA. The results showed that the titer of Ad-Tracker-CTLA4-Ig was 6×1013pfu／L. Soluble fusion protein CTLA4-Ig was expressed in LO2 cells infected with Ad-Tracker-CTLA4-Ig for 72 hours(P/N=4.6). It was suggested that Ad-Tracker-CTLA4-Ig can efficiently transfer exogenous gene into LO2 cells and induce the expression of protein CTLA4-Ig. It may be used as a novel immunosuppressive agent for gene therapy in organ transplantation.

肝移植是治疗终末肝硬化的唯一有效方法。然而移植后乙型肝炎复发严重影响其治疗效果。我们采用重组腺相关病毒介导人乙型肝炎病毒表面抗体，使其特异表达于肝细胞内。观察其表达产物在树鼯中预防乙型肝炎病毒感染的作用，以了解该系统在移植肝中预防乙型肝炎病毒再感染的作用。通过本研究可望用于肝移植中治疗肝硬化预防乙型肝炎病毒复发。