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目的 在心脏移植手术中实施CTLA 4Ig腺病毒基因治疗并联合术后输注供体骨髓细胞 ,诱导异基因大鼠心脏移植耐受 ,并对相关机制进行研究。方法 将异基因DA大鼠的心脏移植给受体LEW大鼠 ,同时经门静脉输注供体DA的脾细胞 (SC ,3× 10 8)、CTLA 4Ig腺病毒 [( 1～ 5 )× 10 9PFU ml],第 4天由舌静脉输注DA的骨髓细胞 (BMC ,3× 10 8)。观察、记录心脏移植物的存活时间。并对皮肤移植的受体作同样处理 ,观察皮肤移植存活情况。通过MLR、IL 2逆转实验及嵌合体的测定 ,探讨耐受机理 ,并检测了CTLA 4Ig的体内表达、TH1 TH2型细胞因子的表达。结果 单用CTLA 4Ig腺病毒基因治疗 ,或CTLA 4Ig腺病毒基因治疗联合单独的供体脾细胞或骨髓细胞能不同程度地延长异基因心脏移植物的存活 ,但不能延长皮肤移植物的存活。脾细胞、CTLA 4Ig腺病毒和骨髓细胞(SC Ad BMC)处理组的心脏移植物存活时间明显超过其它各耐受诱导组 ,并且能够诱导皮肤耐受。RT PCR实验证明 ,在受体内不同的组织CTLA 4Ig基因的表达量有所不同 ,并且随着时间的推移表达下降。TH1和TH2型细胞因子的检测显示 ,耐受大鼠体内未发现这两类细胞因子的偏移现象。MLR证明耐受大鼠的免疫应答表现为供体特异性降低 ,IL 2逆转实验、嵌合体检测表明 ,该耐受可能与?
Objective To perform gene therapy of CTLA 4Ig adenovirus during heart transplantation and to combine donor and donor bone marrow cells to induce allograft tolerance of allogeneic rats. The related mechanisms were studied. Methods The heart of allogeneic DA rats was transplanted to the recipient LEW rats. At the same time, donor spleen cells (SC, 3 × 10 8), CTLA 4Ig adenovirus [(1 ~ 5) × 10 9 PFU ml], and DA bone marrow cells (BMC, 3 × 10 8) were infused into the tongue vein on the fourth day. Observe and record heart graft survival time. And the skin graft recipients for the same treatment, observation skin graft survival. Tolerance mechanism was investigated by MLR, IL 2 reversal assay and chimerism assay. The expression of CTLA 4Ig in vivo and the expression of TH1 TH2 cytokines were detected. Results Single CTLA 4Ig adenovirus gene therapy, or CTLA 4Ig adenovirus gene therapy alone donor spleen cells or bone marrow cells can extend the survival of allogeneic heart allografts to varying degrees, but can not prolong the survival of skin grafts. Cardiac graft survival time of splenocytes, CTLA 4Ig adenovirus and bone marrow cells (SC Ad BMC) treatment group significantly exceeded that of other tolerance induction groups and was able to induce skin tolerance. RT PCR experiments show that different tissues in the recipient CTLA 4Ig gene expression levels vary, and with the passage of time decreased expression. The detection of TH1 and TH2 cytokines showed that no shift in these two types of cytokines was found in the tolerant rats. MLR demonstrates that immune responses in tolerant rats appear to be donor specific decreases, IL 2 reversal assays, and chimera tests indicate that this tolerance may be related to?