体外扩增自体CD4^+CD25^+调节性T细胞促进小鼠供体来源肿瘤的转移  

Ex vivo expanded autologous CD4^+CD25^+ regulatory T cells (Tregs) promote donor derived tumor metastasis in mice

作　　者：刘钊[1] 罗诗樵[1] 郭涛[1] 高祥[1] 

机构地区：[1]重庆医科大学附属第一医院肝胆外科,400016

出　　处：《免疫学杂志》2014年第10期845-849,共5页Immunological Journal

基　　金：重庆市自然科学基金计划项目(cstc2013jcyjA10105);重庆市医学科技计划项目(2008-2-02)

摘　　要：目的研究输注体外扩增自体CD4+CD25+调节性T细胞(regulatory T cells,Tregs)治疗是否存在增加供体来源肿瘤转移的风险。方法取Balb/c(H-2d)小鼠骨髓细胞,在重组鼠源性GM-CSF和IL-4诱导以及TNF-α刺激下分化为成熟树突状细胞。免疫磁珠法(MACS)从Balb/c(H-2d)小鼠脾脏分选出CD4+CD25+Tregs,加入成熟树突状细胞和高浓度重组鼠源性IL-2,体外扩增7 d。流式细胞术检测新鲜以及扩增后CD4+CD25+Tregs纯度及免疫抑制功能。将Balb/c(H-2d)小鼠随机分为2组,实验组:经尾静脉注射1×107体外扩增后CD4+CD25+Tregs,24 h后注射5×105 B16-F10(H-2b)(鼠源性黑色素瘤细胞);对照组:单独输注5×105 B16-F10(H-2b)。结果新鲜分选和扩增后CD4+CD25+Tregs纯度及免疫抑制功能无明显下降。对照组肺部肿瘤结节为(9±8)个,实验组肺部肿瘤结节为(118±15)个。与对照组相比,实验组肺部肿瘤结节明显增加(P<0.01)。结论体外扩增的CD4+CD25+Tregs具有抑制机体抗肿瘤能力,过继输注体外扩增的自体CD4+CD25+Tregs治疗移植后排斥反应,能够增加供体来源肿瘤转移的风险。Adoptive transfer of in vitro expanded autologous CD4＋CD25＋Tregs to prevent rejection is an attractive immunosuppression strategy in transplantation.However,inhibition of anti-tumor immunity by Tregs had been proved in clinical reports,which raised concerns on infusing Tregs into recipient.This study performed to test whether adoptive transfer of ex vivo expanded Tregs would inhibit anti-immunity of recipients.Mature recipient dendritic cells（H-2d） were generated through inducing bone marrow cells of Balb/c（H-2d） mice in the presence of recombinant murine GM-CSF,IL-4 and TNF-α;CD4＋CD25＋Tregs were sorted by MACS from Balb/c（H-2d） mice,and some Tregs were expanded ex vivo with recombinant murine IL-2 and mature dendritic cells for 7 days.Dendritic cells and purity of regulatory T cells were tested by flow cytometry.In vitro inhibition of CD4＋CD25-T cell response by fresh isolated or expanded Tregs was measured by CFSE dilution mixed lymphocyte reaction test.Balb/c（H-2d） mice were randomly divided into two groups：in experimental group,Balb/c（H-2d） mice were firstly inoculated with 1×107expanded CD4＋CD25＋Tregs,and 5×105B16-F10（H-2b） murine melanoma cells were injected intravenously after 24 hours;in control group,mice were inoculated with 5×105B16-F10（H-2b） murine melanoma cells alone.After 14 days,mice were sacrificed and the black tumor nodules in lungs were counted.Result showed ex vivo expanded Tregs inhibited CD4＋CD25-T cells response,as compared to fresh isolated Tregs.Furthermore,there were 9±8 pulmonary tumor nodules in control group,and 118±15 in experimental group.Compared with the control group,the pulmonary tumor nodules were significantly increased in experimental group（P〈0.01）.In conclusion,in vitro expanded autologous CD4＋CD25＋Tregs can suppress anti-tumor immunity,thus the increased risk of donor derived tumor should be considered before their application in rejection suppression.

关 键 词：调节性T细胞 成熟树突状细胞 肿瘤 

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