肝移植术后排斥严重影响患者生存，目前调控Breg分化已成为减轻肝移植排斥反应的热点。在前期研究中我们发现GSK-3β可调节Treg分化和功能减轻排斥反应，还发现急性排斥患者外周血Breg与Treg同步减少，呈正相关，且Breg的GSK-3β活性升高，体外抑制GSK-3β使B细胞向Breg分化增多，并增强Breg抑制CD4+CD25-T细胞的增殖和促进naïve CD4+T细胞向Treg分化。为此提出假说：抑制GSK-3β活性促进Breg分化，并增强Breg调节naiveCD4+T细胞和肝内Kupffer等免疫细胞的分化和功能促进移植肝免疫耐受。本项目将建立大鼠肝移植模型，采用流式细胞仪、病毒载体转染等手段，从分子、细胞、动物模型等方面研究，明确GSK-3β调控Breg分化进而影响移植肝内外多种免疫细胞分化和功能，最终诱导移植肝免疫耐受的机制。项目成功实施将为提高肝移植长期疗效提供新策略。

Liver transplantation has become a lifesaving procedure for patients with chronic end-stage liver disease. However, rejection remains a major cause of early graft loss and a risk factor that impairing long-term recipient post-transplant survival. Recent studies have shown the critical role of regulatory B cells(Breg) in regulating immunological response of transplantation. We conducted a preliminary study and found that GSK-3β could regulate Treg differentiation and function leading to reduced acute rejection. We also found that the proportion of Breg in the peripheral blood was lower in patients with acute rejection as compared with that of non-acute-rejection patients. Meanwhile, higher levels of GSK-3β activation in Breg cells were found in patients with acute rejection. Moreover, GSK-3β inhibition by SB216763 significantly enhanced Breg differentiation, resulting in increased Treg differentiation from CD4+ naive T cells by Breg co-culture in vitro. We hypothesize that inhibition of GSK-3β promotes Breg differentiation and enhances their regulation of naive CD4 + T cell and Kupffer cell differentiation and immune response in the liver graft, leading to alleviated graft rejection and better transplant tolerance after liver transplantation. To verify this hypothesis, we plan to take advantages of lentiviral vector transduction, RNA interference and molecular inhibitors for interference of GSK-3β activation to investigate the roles of GSK-3β in the liver transplant tolerance both in vivo and vitro. This project could provide new ideas and strategies for improving the patient long-term outcome after liver transplantation.