摘要
目的研究过表达CC趋化因子受体7(CCR7)的C3H10 T1/2小鼠间充质干细胞系在小鼠皮肤移植模型中,是否具有比原始C3H10 T1/2细胞更强的免疫抑制作用。方法培养扩增稳定的小鼠C3H10 T1/2细胞,并利用慢病毒工具导入EGFP-CCR7基因;以C57BL/6小鼠为供体获取皮片,以BALB/c小鼠为受体建立小鼠同种异系皮肤移植模型;实验动物分4组:CCR7+C3H10 T1/2细胞组、C3H10 T1/2组、同种异系对照组和同系对照组(受者小鼠移植同系小鼠皮片),分别于移植模型成功当天,经尾静脉注射1×106C3H10 T1/2细胞、1×106CCR7+C3H10 T1/2细胞、等体积生理盐水(对照组);观察皮片移植后的存活情况,并于皮片移植第12天,测定小鼠脾淋巴细胞中Th17细胞与调节性T细胞(Treg)的比例,HE染色观察皮片病理变化。结果受者小鼠皮片移植后12 d,外观、组织病理形态观察以及流式细胞术检测结果表明,CCR7+C3H10 T1/2细胞与C3H10 T1/2细胞的皮片存活情况均优于同种异系对照组;CCR7+C3H10 T1/2细胞皮片移植后存活情况优于C3H10 T1/2细胞组,且移植免疫反应程度轻于C3H10 T1/2细胞组。结论 CCR7+C3H10 T1/2细胞相对于正常的C3H10 T1/2细胞,拥有更好的免疫抑制效应,可有效的降低移植免疫炎性反应,改善移植物的生存状态。
Objective To evaluate whether the chemokine （ C-C motif） receptor 7 （CCR7） over-expressing C3H10 T1/2 mesenchymal stem cells are more efficient in immunosuppression effects than the original ones in skin grafts in mice. Methods The C3H10 T1/2 cells were amplified and transfered with EGFP-CCFr7 gene using lentivirus. The skin grafts were harvested from donators （C57BL/6 mice） and then transplanted to recipients （BALB/c mice）. The recipients were divided into 4 groups randomly： CCR7^＋ C3H10 T1/2 group （1 × 10^6 CCR7^＋ C3H10 T1/2 cells suspended in 1 mL PBS were injected into the recipient mice by caudal vein）, C3H10 T1/2 group （ 1 × 10^6 C3H10 T1/2 cells were injected into the recipient mice after skin graft）, allogenic control group （the mice were injected with 1 mL normal saline）, and cengenic centrol group （the BALB/c mice that had received the skin from the congenic ones were injected with 1 mL normal saline）. The skin graft survival condition and hietopathological changes were observed on the 12th postoperative day. The Th17 cells and regulatory T cells （Tregs） in the lymphocytes of spleen were detected by mouse Th17/Treg phenotyping kit. Results On the 12th postoperative day, the skin surface features, histopathologic changes and flow cytometry results indicated that the skin graft survival condition of the allografts treated with CCR7^＋ C3H10 T1/2 or C3H10 T1/2 cells were better than that of the allogenic control group. Compared with C3H10 T1/2 cells, CCR7^＋ C3H10 T1/2 cells had better skin graft survival condition and weaker immune response. CondeBion CCR7^＋ C3H10 Tl/2 cells induce more intensive immunosuppressive effects than odginal C3H10 T1/2 cells. CCR7^＋ C3H10 T1/2 cells significantly inhibit inflammatory reaction and improve the living state of skin allograft.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2015年第7期873-878,共6页
Chinese Journal of Cellular and Molecular Immunology
