F Protein In Liver Transplantation Immune Tolerance Induced By A Preliminary Study

With the continuous progress of liver transplantation and improve, making more and more the survival of patients with end-stage liver disease has been a large degree of extension. However, acute rejection (Acute Rejection, AR) after liver transplantation remains the most serious complication, its incidence remains high at 40%-60%, so immune tolerance is still the focus of the study of liver transplantation. There are many mechanisms of immune tolerance theory, a variety of effective immunosuppressive agents are constantly introduced, but more and more attention in recent years, the liver specific protein (F protein) induction of immune tolerance after liver transplantation research, so that we on the occurrence of liver transplant tolerance, the development has been further understood.F protein expression and purificationIn this paper, based on previous studies, the F of prokaryotic expression vector pET-15b expression into E. coli strain BL21 (DE3) pLysS, induced with IPTG expressed by affinity chromatography after the (Ni2+-charged IDA His-Bind column) and purified using sodium dodecyl sulfate-gel electrophoresis and polypropylene amine phthalocyanine immunoblotting (Western blot) to detect them. F protein expression in human strains BL21 (DE3) pLysS after the expression of whole-cell protein SDS-PAGE electrophoresis, molecular size of the target band of approximately 43kD, consistent with the literature reports. A western blot showed the purified F protein and F protein purified guinea pig anti-human polyclonal antibody response was positive, that we obtained by genetic engineering of the F protein purified immune activity.F protein in liver transplantation immune tolerance induced by a preliminary studyTo observe the acute rejection and the immune tolerance after orthotopic liver transplantation in rats between inbred line Wistar and closed colony SD. We established models with Kamada's two cuff technique and then randomly divided into three groups:Group A:SDâ†'SD;Group B:Wistarâ†'SD; Group C:F protein intervention:intrathymic injection one week before surgery. Three recipients were killed after 3d,6d,9d and 100d. Representative liver allografts tissue and serum specimens were collected for corresponding detection. Remaining recipients were observed for long-term survival. During the experiment 108 models were established, the total success rate was 62.96%(68/108). The success rate was only 24.44%(11/45) during the protophase of model manufacture. Afterward the success rate gradually comes up to more than 90%.The causes of death were hemorrhage, anesthetic accident, long anhepatic phase, thrombus formation and so on. In recipients of A group, rejection was not founded. In recipients of B group, I grade rejection occurred at 3rd day and rejection gradually reached the peak 6 days after transplantation, and most died between 12 to 21 days after operation. In recipients of C group rejection was not founded either. Conclusions:(1) Two cuff technique may provide a practical and stable experimental model.(2)The keys of establishing successful models include high quality donor liver, practiced microsurgical techniques and short anhepatic phase.(3) The model can be used to study acute rejection or immunologic tolerance effectively.(4) F protein can induce immune tolerance in liver transplantation.