NKT细胞在诱导小鼠门静脉耐受中的作用
李卓男1，关连越1，刘宏宇1，辛敏刚2，王展鹏1，叶研硕1，付佩尧1，李巍1
基金项目：基金项目：国家自然科学基金（81170416; 81273264）； 教育部博士点基金(20100061110069）；吉林省科技厅国际合作基金（2011742）；天普研究基金（01201046)；吉林省自然科学基金（201015178） 作者简介：李卓男（1987年生），男，经治医师，器官移植与免疫耐受 通信联系人：李巍（1963年生），女，教授，器官移植与免疫耐受.

（1. 吉林大学中日联谊医院肝胆胰外科，长春，130033； 2. 吉林大学中日联谊医院麻醉科，长春，130033）
摘要：目的：明确NKT细胞在介导门静脉耐受中的作用。方法：利用小鼠同种异体异位心脏移植模型，移植前经门静脉注入同种异体脾细胞诱导心脏移植物免疫耐受，经尾静脉注入同种异体脾细胞组设为对照，观察两组心脏移植物存活情况，应用流式细胞术方法监测肝脏非实质细胞的亚型的变化，应用免疫组化等方法分析淋巴细胞免疫功能。结果：同种异体抗原经门静脉注入后肝脏中NKT细胞数量明显增加，并表达的较高的PD-L1水平，同时CD4+CD25+Foxp3+调节性T细胞（Treg）在受体肝脏及脾内均明显增加，并表达较高的CTLA4、PD-1和PD-L1分子。心脏移植物存活时间在经门静脉注入同种异体抗原的实验组受体鼠内明显延长。结论：NKT细胞在诱导门脉免疫耐受中起重要作用，经门静脉注入同种异体抗原能够诱导肝脏内NKT扩增，继而诱导受体鼠肝及脾内CD4+ CD25+ Foxp3+ Treg15 细胞扩增，显著延长同种异体心脏移植物存活时间。
关键词：NKT细胞；调节性T细胞；小鼠；心脏移植；门脉耐受
中图分类号：R392.4
Effects of NKT cells on portal tolerance induction
Li zhuonan1, Guan lianyue1, Liu hongyu1, Xin mingang2, Wang zhanpeng1, Ye yanshuo1, Fu peiyao1, Li wei1
(1. Dept of Hepatobiliary-pancreatic Surgery,China-Japan Union Hospital of Jilin University,Changchun,130033; 2. Anesthesiology,China-Japan union hospital of Jilin University,Changchun,130033)

Abstract: Aim: to explore the role of NKT cells on portal tolerance induction. Method: allogeneic mouse heterotopic heart transplantation model was employed. Allogeneic spleen cells was injected through the portal vein before heart transplantation, the spleen cells injected through the tail vein was used as control. Heart allograft survival was followed and the phenotypes of liver nonparenchymal cells were characterized by flowcytometry. The cytokine profiles of host lymphocytes were detected by  immunohistochemistry. Results: liver NKT cells was increased significantly after receiving the allogeneic spleen cells through portal vein, rather than tail vein injection. PD-L1 level were also increased on NKT cells. CD4+CD25+Foxp3+ regulatory T (Treg) cells were increased too in the livers and spleens of the mice who received alloantigen injection through portal vein. Those Treg expressed high CTLA4, PD1 and PD-L1 molecules on their surface. The heart allograft survival time was significantly prolonged in the mice who received alloantigen through portal vein injection. Conclusion: NKT cells plays an important role on portal tolerance induction. Alloantigen delivery through the portal vein results in NKT cells expansion in the liver and CD4+CD25+Foxp3+ Treg expansion in both livers and spleens, and further prolongs the heart allograft survival.
Key words: NKT cells；Treg cells；mouse；heart transplantation；portal tolerance