摘要
目的探讨从转染IKK2dn并负载供者抗原的未成熟树突状细胞（imDC）诱导产生的调节性T细胞（Treg）中筛选CD4＋CD25-Treg的方法，并进行鉴定。方法Lewis大鼠骨髓源性imDC，转染IKK2dn后负载供者BN大鼠抗原，与Lewis大鼠T细胞进行体外混合淋巴细胞反应（MLR）诱导产生Treg，用免疫磁珠法（MACS）筛选出CD4＋CD25-T细胞，流式细胞仪（FCM）检测细胞纯度。加入CD4＋CD25-T细胞行再次MLR检测其抑制T细胞增殖的作用。结果经MACS筛选，CD4＋CD25-T细胞纯度为（95．78±1．25）％。再次MLR结果显示CD4＋CD25-T细胞组的吸光度值为（0．106±0．006），低于BN抗原组（0．189±0．007）、Adv0—CD4＋T细胞组（0．419±0．014）及第三方供者抗原组（0．200±0．008），差异有统计学意义（P〈0．05）。结论转染IKK2dn并负载抗原的imDC诱导产生的Treg，经MACS筛选可以获得高纯度的CD4＋CD25-T细胞，对同种T细胞增殖具有针对供者的特异性免疫抑制作用。
Objective To investigate the method of screening CD4 ＋ CD25 - T cells from regulatory T cells （Treg） induced by recipient-derived immature dendritic cells （imDC） transfected by IKK2dn and loaded with donor antigens, and assess their immunologic function. Methods Rat bone marrow-derived imDC were transfected by IKK2dn and loaded by BN antigen, then cultured with Lewis rats T cells in vitro. From these Treg, CD4＋ CD25-T cells were screened by magnetic active cell sorting （MACS）, then incubated in secondary mixed lymphocyte reaction （ MLR）, with Lewis rat T lymphocytes. Results By MACS, the purity of CD4＋ CD25-T cells was more than 95%. The result of secondary MLR displayed the absorbance value was significantly lower in the CD4＋ CD25 - T cells group （0. 106 ± 0. 006 ） than that in BN antigen group （0. 189 ±0. 007）, Adv0-CD4＋ T cells group （0. 419 ±0. 014） and the third donor group （0. 200 ± 0. 008） （P 〈 0. 05）. Conclusion By MACS screening, we can obtain high purity of CD4 ＋ CD25 T cells from Treg induced by imDC transfected with IKK2dn and loaded by BN rat antigen and recipient Lewis rats T cells. And these T cells can inhibit T cell proliferation with the donor antigen specificity.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第6期1076-1079,F0003,共5页
Chinese Journal of Experimental Surgery
基金
基金项目：国家自然科学基金资助项目（K112214710）
