Nrp1^＋T细胞对淋巴细胞增殖的调节作用以及对同种异体移植物的影响  

Effects of Nrp1^+ T cells on the regulation of lymphocyte proliferation and survival time of allograft

作　　者：蔡明[1] 刘畅[1] 周鸿敏[2] 裴向克[1] 石炳毅[1] 陈忠华 

机构地区：[1]解放军第309医院全军器官移植中心泌尿外科,北京100091 [2]华中科技大学同济医学院附属同济医院心胸外科 [3]华中科技大学同济医学院附属同济医院器官移植研究所,教育部、卫生部重点实验室

出　　处：《解放军医学杂志》2009年第11期1276-1280,共5页Medical Journal of Chinese People's Liberation Army

基　　金：全军医学科研“十一五”计划科技攻关课题资助项目(06G115);国家科技支撑计划课题(2008BAI60B04)

摘　　要：目的探讨表达神经纤毛蛋白质1(Nrp1)的T细胞(Nrp+T细胞)对体内外反应性淋巴细胞增殖的调节作用以及对同种异体移植物的影响。方法采用流式细胞术分选近交系C57BL/6j小鼠脾脏中的Nrp1+T细胞作为调节细胞,在刀豆蛋白A(Co-nA)的刺激下与标记了羧基荧光素乙酰乙酸(CFSE)的C57BL/6j小鼠脾脏单个核细胞等比例共培养5d,对比观察反应细胞的增殖情况。采用流式细胞术分选C57BL/6j小鼠脾脏的Nrp1+T细胞、CD4+CD25+Treg细胞和CD4+CD25-T细胞各1×106个,分别经尾静脉注入到以Balb/C小鼠作为供者,以C57BL/6j小鼠作为受者行皮肤移植的小鼠体内,另外建立供、受体均为C57BL/6j小鼠的同品系对照组(注入等体积的RPMI1640培养基)。对比观察各组移植皮肤的存活情况。结果加入Nrp1+T细胞的实验组增殖指数为50.92%±2.30%,明显低于未加入调节性T细胞的对照组(90.31%±1.83%,P=0.0169)。实验组移植皮肤存活时间注入Nrp1+T细胞者为20.66±1.11d,注入Nrp1-T细胞者为13.20±0.76d,注入CD4+CD25+Treg者为17.67±1.0d,均明显高于对照组(9.70±1.23d,P<0.01)。结论Nrp1作为一种新型的T细胞表面分子,在淋巴细胞增殖体系中起着重要作用;与注入CD4+CD25+Treg相比,注入Nrp1+T细胞可明显延长小鼠移植皮肤的存活时间。Objective To investigate the effects of Nrp1＋ T cells on the regulation of lymphocyte proliferation and the effect on prolongation of the survival time of skin allograft. Methods Nrp1＋T cells were sorted from the splenocytes of C57BL/6j mouse by flow cytometry and to serve as responder cells under stimulation with ConA,they were co-cultured with C57BL/6j from mouse spleen with covalent coupling dye CFSE marker for 5 days. The Proliferation Index was quantified by MODFITTM software. Skin transplantation models were reproduced with the C57BL/6j mice as recipients and the BALB/c mice as donors. Nrp1＋ T cells,C57BL/6j CD4＋CD25＋T cells and CD4＋CD25-T cells （1×106） sorted from spleen of C57BL/6j mice were injected via tail vein at the moment when skin transplantation finished. Control was set up with C57BL/6j mice as both recipients and donors. The survival time of skin allograft was compared. Results The Proliferation Index of the experimental group was 50.92%±2.30%,which was significantly lower than that in control group （90.31%±1.83%,P=0.016 9）. Survival time of skin allograft of Nrp1＋ T cells group was 20.66±1.11d,that of Nrp1-T cells group was 13.20±0.76d,and that of CD4＋CD25＋ T cells group was 17.67±1.00d. All the three the survival time of all the three groups was different from that of control group （9.70±0.53d,P0.01） in which the animals were injected with PRIM-1640. Conclusion The sorted Nrp1＋ T cells can suppress lymphocyte proliferation and prolong the survival time of skin allograft more effectively than CD4＋CD25＋ T cells does.

关 键 词：T淋巴细胞 调节性 皮肤移植 细胞增殖 神经纤毛蛋白质1 

分 类 号：R392.4[医药卫生—免疫学]