中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (15): 2728-2732.doi: 10.3969/j.issn.1673-8225.2012.15.015

• 组织构建与生物活性因子 tissue construction and bioactive factors • 上一篇    下一篇

成熟树突状细胞和转化生长因子β联合体外扩增小鼠调节性T细胞*☆

范莎莎1，罗荣城2，段华新3，石詠中1，袁友红1，肖佩玲3   

  1. 1湖南省人民医院临床医学研究所，湖南省长沙市   410005；2南方医科大学附属南方医院肿瘤中心，广东省广州市   510000；3湖南省人民医院血液肿瘤科，湖南省长沙市  410005
  • 收稿日期:2011-10-07 修回日期:2011-11-15 出版日期:2012-04-08 发布日期:2012-04-08
  • 通讯作者: 肖佩玲，主任医师，湖南省人民医院血液肿瘤科，湖南省长沙市 410005 fresh0225@sina.com
  • 作者简介:范莎莎☆，女，1982年生，湖南省衡阳市人，汉族，南方医科大学在读博士，医师，主要从事肿瘤生物治疗及靶向治疗研究。 fresh0225@126.com 并列第一作者: 罗荣城★，男，1958年生，广东省潮州市人，汉族，1989年解放军第一军医大学小儿血液专业毕业，硕士，主任医师，主要从事肿瘤生物治疗与分子靶向治疗研究。
  • 基金资助:

    长沙市科技局课题(k0902174-31)。

Combination of mature dendritic cells and transforming growth factor beta for in vitro amplification of regulatory T cells in mice

Fan Sha-sha1, Luo Rong-cheng2, Duan Hua-xin3, Shi Yong-zhong1, Yuan You-hong1, Xiao Pei-ling3   

  1. 1Institue of Clinical Medicine, Hunan Provincial People’s Hospital, Changsha  410005, Hunan Province, China; 2Department of Oncology, Nanfang Hospital of Southern Medical University, Guangzhou  510000, Guangdong Province, China; 3Department of Neoplastic Hematology, Hunan Provincial People’s Hospital, Changsha  410005, Hunan Province, China
  • Received:2011-10-07 Revised:2011-11-15 Online:2012-04-08 Published:2012-04-08
  • Contact: author: Xiao Pei-ling, Chief physician, Department of Neoplastic Hematology, Hunan Provincial People’s Hospital, Changsha 410005, Hunan Province, China fresh0225@sina.com
  • About author:Fan Sha-sha☆, Studying for doctorate, Physician, Institue of Clinical Medicine, Hunan Provincial People’s Hospital, Changsha 410005, Hunan Province, China fresh0225@126.com Luo Rong-cheng, Master, Chief physician, Department of Oncology, Nanfang Hospital of Southern Medical University, Guangzhou 510000, Guangdong Province, China
  • Supported by:

    Changsha Municipal Science and Technology Bureau, No. k0902174-31*

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摘要：

背景：目前天然CD4+CD25+Foxp3+调节性T细胞稀少，远远不能满足临床需求。
目的：建立有效的体外培养扩增小鼠CD4+CD25+T细胞体系。
方法：从C57/BL6小鼠骨髓获得的成熟树突状细胞联合不同剂量转化生长因子β，体外扩增同基因或异基因CD4+CD25+T细胞。
结果与结论：经过2周培养，磁珠分选后的CD4+CD25+T细胞的扩增倍数为自然调节性T细胞的(17.3±10.6)倍, 0.2，2 μg/L质量浓度对T细胞的扩增能力最强；经扩增后的T双阳性率为(85.38±1.82)%，CD127阳性率(78.86±0.91)%；体外扩增的调节性T细胞高表达FOXP3为天然调节性T细胞的1.5倍；调节性T细胞体外抑制实验结果表明：调节性T细胞对自体或异体CD4+T细胞抑制能力随着效靶比浓度的降低而降低，体外扩增的CD4+CD25+T细胞对自体CD4+T细胞抑制率(60.1±0.71)%，对异体CD4+T抑制率为(35.5±0.57)%(P < 0.05)。提示在体外成功扩增了CD4+CD25+T细胞，并且具有免疫抑制功能，此方法扩增细胞数量多，细胞纯度有很大提高，方法简便。
关键词：成熟树突状细胞；CD4+CD25+T细胞；转化生长因子β；体外扩增；小鼠
缩略语注释：TGF-β: Transforming growth factor beta，转化生长因子β
doi:10.3969/j.issn.1673-8225.2012.15.015

关键词: 成熟树突状细胞, CD4+CD25+T细胞, 转化生长因子&beta, 体外扩增, 小鼠

Abstract:

BACKGROUND: At present, the nature CD4+CD25+Foxp3+ regulatory T cells are too rare to meet the clinical requirement.
OBJECTIVE: To establish an effective amplification system of murine CD4+CD25+T cells in vitro.
METHODS: Mature dendritic cells from C57/BL6 mouse bone marrow were combined with different doses of transforming growth factor β were used to amplify autologous or allogeneic CD4+CD25+T cells in vitro.
RESULTS AND CONCLUSION: After 2 weeks culture, magnetic bead sorted CD4+CD25+T cells in the amplification factor for natural regulatory T cells were (17.3±10.6) times, 0.2 and 2 μg/L concentration had the strongest amplification on T cells. The expansion CD4+CD25+ positive rate was (85.38±1.82)%, CD127 positive rate was (78.86±0.91)%; The expansion of regulatory T cells with high expression of FOXP3 mRNA was 1.5 folds than nature ones in vitro. The regulatory T cells inhibitory test showed that regulatory T cells could inhibit the proliferation of auto-and allo-CD4+T cells in vitro in a cell dose-dependent manner. The inhibit rate of CD4+CD25+T cells to autologous CD4+T cells was (60.1±0.71) %, the inhibit rate of allogeneic CD4+T cells was (35.5±0.57)% (P < 0.05). We successfully amplified CD4+CD25+T cells in vitro. The cells expanded have inhibitory effects in vitro, and as the method can obtain large quantity and purity cells, which may be used in clinical trials in future.
Fan SS, Luo RC, Duan HX, Shi YZ, Yuan YH, Xiao PL. Combination of mature dendritic cells and transforming growth factor beta for in vitro amplification of regulatory T cells in mice. Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(15): 2728-2732.    
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