调节性T细胞培养及输入慢性阻塞性肺病大鼠后免疫细胞变化

摘要

Objective To study the change of immune factor in COPD rats after intravenous injection of regulatory T cells.Methods Twenty-one SPF rats was divided into three groups at random,rat COPD model was built by smoking.We used magnetic bead isolation technic to separate CD4+CD25+ regulatory T cells.Regulatory T cells were cultured and injected into rats though rats' caudal vein according to different dose,5 × 104/mL,5 × 105/mL,5 × 106/mL respectively.Flow cytometry was used to analyze cell factors.ELISA was used to analyze IL-6 and CRP.Restlts Adding JJ316 or IL-2 into medium benefited the proliferation of CD4+CD25+ regulatory T cells.On the 20 th day,regulatory T cells CD4+CD25+ proliferation stopped by adding JJ316 or IL-2 respectively.Regulatory T cells were cultured and injected into rats though rat caudal vein according to different dose.The levels of CRP and IL-6 were decreased when rats were injected by CD4+CD25+regulatory T cell after one week.Conclusions Injection of regulatory T cells is helpful to control inflammation progression of COPD,so the increase of regulatory T cells of patients with COPD may decrease inflammation progression of COPD.%目的 在慢性阻塞性肺病大鼠体内输入调节性T细胞,观察其输入调节性T细胞后大鼠体内的免疫细胞变化.方法 清洁级雄性SD大鼠21只,随机分为3组,通过烟熏建立大鼠COPD模型.经磁珠分离大鼠CD4+CD25+调节性T细胞(Treg)进行培养.细胞培养成功后给COPD大鼠尾静脉输注5×104/mL、5×105/mL,5×106/mL各1 m L.通过流式细胞仪检测细胞因子,采用ELISA检查IL-6,C反应蛋白(CRP).结果 培养基中添加JJ316或重组人IL-2均有利于CD4+CD25+调节性T细胞生长,2种方法在培养至20 d左右均出现细胞生长停滞.培养成功的调节性T细胞以不同量输入3组烟熏COPD大鼠体内,输注1周后体内炎症因子检测提示:大鼠出现CRP,IL-6下降.结论 研究提示大鼠体内输入CD4+CD25+调节性T细胞有助于抑制COPD炎症的进展,通过提升COPD体内调节性T细胞量可能减缓COPD炎症的进展.