• 南京醫(yī)科大學(xué)第一附屬醫(yī)院肝臟移植中心 （南京210029）;;
  • 通訊作者： 王學(xué)浩;

目的探索利用磁性細(xì)胞分離(MACS)系統(tǒng)高效、快速地分離大鼠脾臟CD4+CD25+調(diào)節(jié)性T細(xì)胞。
方法采用免疫磁珠兩步法分離大鼠脾臟內(nèi)的CD4+CD25+調(diào)節(jié)性T細(xì)胞, 首先采用“雞尾酒”抗體和抗IgG磁珠陰性分選CD4+ T細(xì)胞，再用抗CD25PE抗體和抗PE磁珠陽性分選獲得CD4+CD25+T 細(xì)胞。分離后的細(xì)胞經(jīng)流式細(xì)胞儀檢測(cè)分離純度； 臺(tái)盼藍(lán)染色檢測(cè)細(xì)胞存活率； 體外增殖實(shí)驗(yàn)檢測(cè)其對(duì)CD4+CD25-T細(xì)胞的免疫抑制作用。
結(jié)果陰性分選獲得的CD4+T細(xì)胞純度為（83.6±2.5）%（79%~87%）, 陽性分選后獲得的CD4+CD25+T細(xì)胞純度為（90.2±1.8）%（86%~93%）, 細(xì)胞存活率為（92.8±3.4）%（92%~95%）, 體外增殖實(shí)驗(yàn)表明，CD4+CD25+T細(xì)胞能明顯抑制CD4+CD25-T 細(xì)胞的增殖（P＜0.01）。
結(jié)論采用MACS系統(tǒng)陰性加陽性分選可以高效、快速地獲得理想純度和有免疫抑制功能的大鼠CD4+CD25+調(diào)節(jié)性T細(xì)胞。

引用本文： 浦立勇,王學(xué)浩,李相成,張峰,俞悅,姚愛華,成峰,李國強(qiáng). 免疫磁珠兩步法分離大鼠脾臟CD4+CD25+調(diào)節(jié)性T細(xì)胞. 中國普外基礎(chǔ)與臨床雜志, 2006, 13(3): 275-278. doi: 復(fù)制

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  4. 4. [Ｊ]. Int Immunol, 1998; 10(12)∶1969.
  5. 5. [Ｊ]. Blood, 2002; 99(10)∶3493.
  6. 6. [Ｊ]. Blood, 2003; 102(6)∶2180.
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  9. 9. Stephens LA, Barclay AN, Mason D. Phenotypic characterization of regulatory CD4+CD25+ T cells in rats.
  10. 10. Sakaguchi S, Sakaguchi N, Asano M, et al. Immunologic selftolerance maintained by activated T cells expressing IL2 receptor alphachains (CD25). Breakdown of a single mechanism of selftolerance causes various autoimmune diseases.
  11. 11. SuriPayer E, Amar AZ, Thornton AM, et al. CD4+CD25+ T cells inhibit both the induction and effector function of autoreactive T cells and represent a unique lineage of immunoregulatory cells.
  12. 12. Takahashi T, Kuniyasu Y, Toda M, et al. Immunologic selftolerance maintained by CD25+CD4+ naturally anergic and suppressive T cells: induction of autoimmune disease by breaking their anergic/suppressive state.
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  14. 14. Jiang S, Camara N, Lombardi G, et al. Induction of allopeptidespecific human CD4+CD25+ regulatory T cells ex vivo.
  15. 15. Dieckmann D, Plottner H, Berchtold S, et al. Ex vivo isolation and characterization of CD4+CD25+ T cells with regulatory properties from human blood.
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