比较体外扩增调节性T细胞的两种方法  

A Comparison of Two Methods to Expand Regulatory T Cells(Tregs) In Vitro

作　　者：刘钊[1] 罗诗樵[1] 

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：《中国细胞生物学学报》2015年第5期703-708,共6页Chinese Journal of Cell Biology

基　　金：重庆市自然科学基金计划项目(批准号:cstc2013jcyjA 10105);重庆市医学科技计划项目(批准号:2008-2-02)资助的课题~~

摘　　要：利用免疫磁珠法(immunomagnetic beads,MACS)从BALB/c小鼠脾脏中分别分离出CD4+CD25+Tregs(regulatory T cells)和CD4+T细胞,CD4+CD25+Tregs加入anti-CD3/CD28磁珠与1 000 U/m L IL-2体外扩增14 d;CD4+T细胞加入anti-CD3/CD28磁珠、100 U/m L IL-2和5 ng/m L重组人源性转化生长因子-β(transforming growth factor-β,TGF-β)体外扩增5 d。分析两种方法扩增后Tregs免疫抑制功能及数量。结果显示,两种方法扩增的Tregs免疫抑制功能无明显差别(P>0.05),但是分别从大小相当的BALB/c小鼠脾脏所提取出的细胞中,通过CD4+T细胞扩增5 d可以得到(4.9±0.4)×107 Tregs,而通过CD4+CD25+Tregs扩增14 d只能得到(1.9±0.1)×107 Tregs(P<0.05)。结果发现,通过两种方法扩增得到的Tregs免疫抑制功能无明显差异,但是通过CD4+T细胞扩增Tregs方法所需时间短,而且得到的细胞数量多。CD4＋CD25＋Tregs （regulatory T cells） and CD4＋T cells were isolated from BALB/c spleens by MACS （immunomagnetic beads）, respectively. CD4＋CD25＋Tregs were cultured for 14 d in 96-well plates coated with anti-CD3/CD28 microbeads in the presence of 1000 U/mL of IL-2; then CD4＋T ceils were cultured in 24-well plates coated with anti-CD3/CD28 microbeads in the presence of 100 U/mL IL-2 and 5 ng/mL of recombinant human TGF-β （transforming growth factor-β） for 5 d. The immunosuppression and counts of Tregs expanded by the two methods were analyzed. The experiment results showed that there was no obvious difference in immunosuppression between the two methods of expanding Tregs in vitro （P〉0.05）, but when used the same size BALB/c mice, we can get （4.9±0.4）×10^7 Tregs through CD4＋T cells and only get （1.9±0.1）×10^7 Tregs through CD4＋CD25＋Tregs （P〈0.05）. The result suggested that there was no obvious difference in immunosuppression between the two methods of expanding Tregs, but get more Tregs and take less time through CD4＋T ceils.

关 键 词：调节性T细胞 扩增 免疫抑制功能 

分 类 号：Q2-33[生物学—细胞生物学]