CAJ | 학술논문

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.
목적 탐토종전염IKK2dn병부재공자항원적미성숙수돌상세포(imDC)유도산생적조절성T세포(Treg)중사선CD4+ CD25 - Treg적방법,병진행감정.방법 Lewis대서골수원성imDC,전염IKK2dn후부재공자BN대서항원,여Lewis대서T세포진행체외혼합림파세포반응(MLR)유도산생Treg,용면역자주법(MACS)사선출CD4+ CD25 -T세포,류식세포의(FCM)검측세포순도.가입CD4+ CD25 -T세포행재차MLR검측기억제T세포증식적작용.결과 경MACS사선,CD4+ CD25 -T세포순도위(95.78±1.25)％.재차MLR결과현시CD4+ CD25 -T세포조적흡광도치위(0.106±0.006),저우BN항원조(0.189±0.007)、Adv0-CD4+T세포조(0.419±0.014)급제삼방공자항원조(0.200±0.008),차이유통계학의의(P＜0.05).결론 전염IKK2dn병부재항원적imDC유도산생적Treg,경MACS사선가이획득고순도적CD4+CD25-T세포,대동충T세포증식구유침대공자적특이성면역억제작용.
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 sereening,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.