T细胞疫苗对肝脏移植物内浸润细胞凋亡的影响  

Effects of inoculation of T cell vaccine on apoptosis of GICs after orthotopic liver transplantation in rats

作　　者：赵振林[1] 吴永嘉[1] 祝庆华[1] 

机构地区：[1]山西医科大学第二医院普通外科,太原市030001

出　　处：《中华肝胆外科杂志》2006年第5期340-342,共3页Chinese Journal of Hepatobiliary Surgery

摘　　要：目的探讨 T 细胞疫苗对肝脏移植物内浸润细胞凋亡的影响。方法应用近交系 DA大鼠和近交系 Lewis 大鼠作为实验动物。制备 DA 大鼠针对 Lewis 大鼠的特异性 T 细胞疫苗(TCV)。第一组(n=8):以 Lewis 大鼠作为供者,以 DA 大鼠作为受者。第二组(n=8):用 Flt3-L(fms-liketyrosine kinase 3 ligand)治疗供者 Lewis 大鼠,以 DA 大鼠作为受者。第三组(n=8):用 Flt3-L 治疗Lewis 大鼠,用 TCV 免疫受者 DA 大鼠。应用三袖套法进行大鼠原位肝移植,观察肝移植物平均存活时间(MST);于移植后第5天获取肝移植物制作组织切片,HE 染色,光镜观察排斥反应情况。同时分离肝脏移植物非实质细胞(NPC),应用流式细胞仪对移植物浸润细胞(GICs)的凋亡和 CD4^+T细胞和 CD8^+T 细胞进行检测。结果第一组进行单纯移植,平均存活时间(MST)为(17±2.6)d、第二组用 Flt3-L 治疗供者后 MST 下降为(5.7±1.4)d(P<0.01)、第三组是在第二组的基础上用 TCV免疫受体诱导耐受,使 MST 显著延长(15.7±4.2)d(P<0.01);移植物组织切片 HE 染色光镜观察所见:第二组可见明显的排斥病理损伤征象,在第一组和第三组未发现明显的排斥性病理改变;GICs凋亡率第一组为(11.94±3.12)%、第二组为(7.92±2.25)%,和第一批比较显著降低(P<0.05),应用 TCV 免疫的第三组 GICs 凋亡率比第二组显著上升,为(25.99±2.49)%(P<0.01);应用 T 细胞疫苗的第三组和单纯移植的第一组其 CD_4/CD_8显著低于第二组(P<0.01)。结论异品系大鼠肝移植物在没有任何免疫抑制治疗的情况下可以被完全接受,供体抗原特异性 TCV 通过诱导肝脏GICs 凋亡抑制 Flt3-L 诱导的肝移植急性排斥反应,GICs 凋亡在同种移植耐受过程中发挥关键作用。Objective To investigate the effects of T cell vaccination on apoptosis of GICs after orthotopic liver transplantation （OLT） in rats. Methods Inbred male DA rats and inbred male Lewis rats were used as the recipients and donors, respectively. T cell vaccine was made from the attenuated spleen cells of DA rats, which was induced by Con A and challenged with the spleen cells of Lewis rats. In group A （n = 8）, OLT was performed using the recipients of DA rats and donors of Lewis rats. In group B （n = 8）, OLT was performed using the recipients of DA rats and donors of Lewis rats treated with Flt3-L. In group C （n = 8）, OLT was conducted using the recipients of DA rats inoculated with T cell vaccine and donors of Lewis rats treated with Flt3-L. The median survival time （MST） of liver grafts was recorded. On the 5^th d after the operation, the rejection was defined by histological observation. Meanwhile, the NPC of liver grafts was isolated so as to determine the apoptosis of GICs, CD4^＋ T cells and CD8^＋ T cells were assayed using flow cytometry. Results The MST were 17±2.6 d, 5.7±1.4d and 15.7±4.2 d in group A, group B and group C, respectively. Histological examination showed that the liver damage was noticeable in group B but the liver grafts had good preservation with no overall architectural changes in group A and group C. Flow cytometry revealed that the percentage of apoptotic GICs was significantly increased in group A and group C as compared with group B （P 〈 0.05）. Conclusions Liver grafts can be completely accepted by the recipient rats under the condition of no immunosuppressive therapy. Donor-Ag specific T cell vaccination can successfully inhibit acute liver graft rejection induced by Flt3-L. The apoptosis of GICs is critical for liver grafts to be accepted.

关 键 词：肝移植 自动耐受 排斥 T细胞疫苗 

分 类 号：R657.3[医药卫生—外科学] R392.7[医药卫生—临床医学]