短期使用雷帕霉素联合调节性T细胞可延长小鼠移植心脏时间但不能抑制受体肿瘤发生  

Short-term use of rapamycin combined with Tregs prolongs mouse cardiac graft survival but attenuates recipient's anti-tumor immunity

作　　者：闫挺[1] 骆晨[1] 杨先模 季磊[1] 罗诗樵[1] YAN Ting LUO Chen YANG Xianmo JI Lei LUO Shiqiao(Department of Hepatobiliary Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China)

机构地区：[1]重庆医科大学附属第一医院肝胆外科,重庆400016

出　　处：《细胞与分子免疫学杂志》2017年第2期174-178,184,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(30972789);重庆市自然科学基金(cstc2013jcyj A10105)

摘　　要：目的研究短期使用雷帕霉素(Rap)联合调节性T细胞(Treg)在诱导同种异体小鼠心脏移植物长期存活,同时探讨此方案对移植受体肿瘤免疫的影响。方法免疫磁珠法分离得到受体小鼠脾脏Treg,经CD3/CD28单克隆抗体磁珠及2000 U/m L重组小鼠白细胞介素2(rm IL-2)体外扩增后,流式细胞术检测纯度;建立小鼠腹腔同种异体心脏移植模型(H-2~b到H-2~d),分为对照组(单纯移植)、Rap组、Rap联合Treg组。Rap组连续14 d经腹腔注射Rap 1 mg/(kg·d),Rap联合Treg组注射相同剂量的Rap,并移植当天经尾静脉过继输注扩增后的1×107个Treg,同时设同基因移植组(H-2~d到H-2~d),每日观察移植心脏搏动并在相应时点进行组织学评价。在受体肿瘤免疫实验中设三组同种异体心脏移植(H-2~b到H-2~d)小鼠,同时经尾静脉过继输注B16-F10细胞(H-2~b)(受体来源),另三组同种异体心脏移植(H-2~d到H-2~b)小鼠经尾静脉过继输注B16-F10细胞(H-2~b)(供体来源),两周后比较肺部肿瘤结节数量。结果对照组移植心脏中位生存时间(MST)为7 d,Rap组为15 d,Rap联合Treg过继输注能够显著延长移植心脏MST至93 d,组织学显示长期存活心脏淋巴细胞浸润及慢性血管病变;对于供体来源肿瘤,对照组未见肿瘤结节,Rap组为(15±8)个,Rap联合Treg组小鼠肺部肿瘤结节为(14±7)个,后2组类似无显著性差异;对于受体来源肿瘤,对照组肿瘤结节为(70±12)个、Rap组为(28±9)个、Rap联合Treg组小鼠肺部肿瘤结节为(146±12)个,与对照组、Rap组相比均显著增加。结论短期使用低剂量Rap联合Treg能够显著延长小鼠移植心脏的存活时间,但仍不能抑制受体肿瘤的发生。Objective To study the effect of short-term use of rapamycin（ Rap） combined with regulatory T cells（ Tregs）on the long-time survival of allogeneic mouse cardiac transplant,and its impact on the anti-tumor immunity of recipient. Methods Mouse Tregs were purified from recipients＇ spleen by magnetic activated cell sorting（ MACS）,and expanded by CD3 / CD28 monoclonal antibody immunomagnetic beads and 2000 U / m L recombinant mouse IL-2（ rm IL-2） ex vivo. The purity was tested by fluorescence-activated cell sorting（ FACS）. Allogeneic mouse cardiac transplanted models were established（ H-2^b to H-2^d）,and the mice were divided into three groups： control group（ transplant only）,Rap group,and Rap combined with Tregs group. In the Rap group,the mice were treated with Rap [1 mg /（ kg·d）,ip] for 14 consecutive days,and the mice in the Rap plus Tregs group received the same treatment,and 1 × 10^7 Tregs were adoptively transferred through the tail vein on the day of transplantion. Meanwhile,the syngeneic transplanted group was set up（ H-2^d to H-2^d）. Allograft survival was monitored daily and the graft was harvested on the indicated day and histologically evaluated. In the experiment of recipient＇s anti-tumor immunity,the similar three groups of allogeneic cardiac transplanted models were established（ H-2^b to H-2^d）,and B16-F10 cells（ recipient derived） were transferred through the tail vein, another three groups of allogeneic cardiac transplanted mice（ H-2^d to H-2^b） were also transferred with B16-F10 cells（ donor derived）. Two weeks later,the tumor nodules of the lung were compared. Results The median survival time（ MST） of the graft was 7 days in the control group,15 days in the Rap group,and 93 days in the Rap combined with Tregs group. Histologic analysis of long-time survival grafts showed lymphocyte infiltration and chronic vasculopathy. For donor-derived tumor,there was no tumor nodule in the control group,and tumor nodules significantly increased to 1

关 键 词：调节性T细胞(Treg) 心脏移植 肿瘤免疫 

分 类 号：R392.4[医药卫生—免疫学] R392.9[医药卫生—基础医学]