SOCS1调控未成熟树突细胞分化诱导肝移植免疫耐受及机制研究

目的探讨细胞因子信号传导抑制因子-1(Suppressors of cytokine signaling-1,SOCS1)基因腺病毒转染未成熟的树突细胞(Dendritic cells,DC),并免疫受体小鼠,通过小鼠肝移植模型,观察免疫耐受的效果并探讨可能的机制。方法构建SOCS1腺病毒并PCR鉴定,感染体外分离、培养的小鼠骨髓树突细胞,Westernblot法检测SOCS1表达。以未转染组及空白转染组为对照,免疫肝移植小鼠模型,检测受体小鼠脾脏淋巴细胞混合淋巴细胞反应(MLR),检测血清IL-2和IFN-γ水平及肝脏组织IL-2和IFN-γmRNA表达。结果成功构建并鉴定转染腺病毒,感染DC后,SOCS1蛋白表达水平较未转染组及空白转染组显著增高(P<0.05)。SOCS1基因修饰的DC免疫肝移植模型小鼠后,受体小鼠脾脏淋巴细胞MLR较未转染组及空白对照免疫受体组小鼠显著下降(P<0.05);SOCS1-DC免疫受体小鼠血清IL-2和IFN-γ水平及肝脏mRNA表达水平均显著下降(P<0.05)。结论通过腺病毒转染使SOCS1在DC中过度表达,用以体内免疫同种异体肝移植受体小鼠,能有效减轻同种移植排斥反应,并在一定程度上诱导形成免疫耐受。

Objective To study the effect on immune tolerance in heterotopic liver transplantation mode mice of Cytokine Signaling-1 （SOCS 1） gene modified dendritic cells （DC）. Methods The adenovirus vectors containing suppressors of cytokine signaling-1 （SOCS1） was constructed. BALB/c mice was immunized with imDC, DC-GFP and DC-SOCS1, the serum level and the mRNA expression in liver of IL-2 and IFN-y was detected. And mixture lymphocyte reaction （MLR） in spleen lymphocytes was observed in BALB/c mice. Results The adenovirus vectors containing SOCS1 was successfully constructed and tranfected. BALB/e mice immunized by DC-SOCS1 exhibited low reactive in spleen lymphocyte MLR （P 〈 0.05）. Serm IL-2 and IFN-y levels determined by ELISA in the DC-SOCS 1 group increased significantly after liver allograft transplantation,but was lower compared to control group and imDC,DC-GFP group （P〈 0.05）. The semiquantitative reversed transcriptase polymerase chain reaction assay showed that expression levels of IL-2 and IFN-y in DC-SOCS1 mice also decreased markedly compared with other groups （P〈 0.05）. Conclusion Replication defective adenovirus vector SOCS1 can effectively transfect DC, and increase the expression of SOCS1 gene. Immunization with SOCS1 gene modified immature DC in vivo can induce antigen-specific immune tolerance in organ transplantation.