供体脾细胞联合抗CD154输注诱导肺移植免疫耐受、

Combined donor specific transfusion and anti-CD154 therapy achieves airway allograft tolerance

目的探讨供体特异性输注（DST）联合共刺激阻断（DST／抗CDl54）诱导肺移植后闭塞性细支气管炎（OB）免疫耐受机制。方法建立小鼠DST／抗CD154方案诱导的免疫耐受模型。术后15、25、100d取出移植气道检测形态学改变，利用CSME-80鼠cDNA芯片检测15d耐受组和同种异体移植组移植物内基因表达差异，分析差异有统计学意义的基因表达与排斥和耐受之间的关系。选择部分差异表达基因行实时荧光定量逆转录一聚合酶链反应（RT—qPCR），鉴定验证基因芯片的可靠性。结果免疫耐受组OB前期存在大量显著表达差异基因，Rapl信号通路、CD40／CD40L相关及其他T细胞表面分子相关基因、细胞周期、细胞生长等促进纤维增殖的部分基因显著下调表达。部分促炎因子同同种异体移植组一样上调表达。结论Rapl信号通路、CD40／CD40L相关及其他T细胞表面分子相关基因、细胞周期、细胞生长等促进纤维增殖下调表达的部分基因等可能与肺移植慢性排斥反应耐受相关。

Objective To-define the molecular mechanism of obliterative bronchiolitis （ OB ） fol- lowing heterotopic tracheal transplantation during the early stages of tolerance induction by donor specific transfusion and anti-CD154. Methods Tolerance induction was achieved in tracheal allografts from BALB/C to C57BL/6 mice by donor specific transfusion of splenocytes intravenously and intraperitoneal injection of anti-CD154 mAbs. The differential gene expression was detected in tolerant murine heterotopic tracheal allografts by means of gene microarrays. Differential expression of selected genes was confirmed by real-time fluorescent quantitative reverse transcription （ RT-qPCR）. Results Comparative analysis between DST/an- ti-CD154 protocol-induced tolerant and rejected grafts revealed many genes were specifically downregulated in the tolerant allografts, including RAP1 pathway, CD40/CD40L concerned T cell surface molecules and many profibrotic genes, which were upregulated in the rejected allografts. In the tolerant allografts there was also marked expression of a number of genes for proinflammatory factors. Conclusion These results suggest a vital role for RAP1 pathway, CD40/CD40L concerned T cell surface molecules and many profi- brotic genes in inducing airway transplant tolerance in this model.