雷帕霉素联合人脐带血CD4^+ CD25^+调节性T细胞治疗胶原诱导性关节炎小鼠的初步研究  

Preliminary research of treating mouse collagen-induced arthritis by rapamycin combined human cord blood CD4^+ CD25^+ regulatory T cells

作　　者：杨燕[1] 杨懿铭[2] 李振华[3] 杨洁[2] 任亚娜[2] 谢如峰[2] 范华骅[2] 

机构地区：[1]复旦大学附属华山医院,上海200040 [2]上海市血液中心血液工程研究室,上海200051 [3]华东师范大学,上海200241

出　　处：《中国免疫学杂志》2014年第1期67-72,共6页Chinese Journal of Immunology

基　　金：上海市科委基金项目(No.11140902900);上海市公共卫生重点学科建设课题(No.12GWZX0201)

摘　　要：目的:初步探讨雷帕霉素(Rapa)联合人脐带血CD4+CD25+调节性T细胞(Tregs)对胶原诱导性关节炎小鼠的治疗作用。方法:通过体外扩增培养获得人脐带血Tregs;建立牛Ⅱ型胶原诱导的类风湿性关节炎(CIA)小鼠模型;将CIA小鼠随机分为对照组,Tregs治疗组,Rapa治疗组,Tregs与Rapa联合治疗组;小鼠骨关节切片进行Safranin O-fast green染色、甲苯胺蓝染色以及HE染色;ELISA法检测CIA小鼠血浆抗Ⅱ型胶原抗体浓度;流式细胞术检测CIA小鼠脾脏CD4+T细胞亚型;检测Tregs在体外对CIA小鼠Ⅱ型胶原特异性CD4+T细胞增殖的抑制功能;Transwell分隔培养法比较非接触培养与直接接触培养中Tregs对CD4+T细胞增殖抑制作用的差异。结果:Tregs与Rapa联合治疗能够延缓疾病发生,显著降低关节炎评分,减少CIA小鼠关节中软骨的破坏以及炎症细胞浸润,降低CIA小鼠血浆中抗胶原总IgG抗体水平以及脾脏中Th17细胞比例;Tregs在体外能够有效抑制CIA小鼠胶原特异性CD4+T细胞的增殖,这种抑制作用主要可能是通过直接接触抑制途径起作用。结论:雷帕霉素联合人脐带血Tregs治疗能够改善CIA小鼠关节炎症状。Objective： To explore the treatment effects of rapamycin （Rapa） combined human cord blood CD4 ＋ CD25 ＋ regula- tory T cells （Tregs） on collagen-induced arthritis （CIA） mouse model. Methods. Tregs were obtained from in vitro expansion of CD4 ~ CD25 ~T cells from human cord blood. The collagen-induced arthritis （CIA） mouse model was established. The mice were randomly divided into control group, Tregs treatment group, Rapa treatment group, and Tregs and Rapa combined treatment group. The mouse joint sections were stained by Safranin O-fast green, Toluidine blue, and hematein eosin （H＆E）. Type-two collagen specific IgG antibody in CIA mouse plasma was detected by ELISA. The phenotypes of the spleen CD4＋ T cells of CIA mice were identified by flow cytometry. The inhibitory effect of Tregs on type-two collagen specific CD4 ＋ T cells was analyzed in vitro. The contact inhibition and non contact inhibition mechanism of Tregs acting on CD4 ＋ T cell was further studied by Transwell culture method. Results： Tregs and rapamycin combined treatment could significantly delay the disease onset and alleviate the severity of arthritis, reduce the cartilage destruction and inflammatory cell infiltration in CIA mouse joints, decrease the level of total anti-type-two collagen IgG antibody in CIA mouse plasma and the proportion of Thl7 cells in CIA mouse spleen. In addition, the effective inhibition of human cord blood Tregs on the proliferation of collagen specific CD4＋ T cells of CIA mice was more likely aitributed to a direct contact manner. Conclusion： The combination therapy of rapamycin and human cord blood Tregs can alleviate the arthritis symptoms of CIA mice.

关 键 词：人 脐带血 雷帕霉素 CD4+CD25+调节性T细胞 胶原诱导性关节炎 

分 类 号：R457.2[医药卫生—治疗学]