Wang Feng-Jie, Cui Dan, Qian Wei-Dong
Department of Neurology, the First Affiliated Hospital of Bengbu Medical College, Bengbu, China.
Department of Neurology, Bengbu First People's Hospital, Bengbu, China.
Cell Physiol Biochem. 2018;47(1):390-402. doi: 10.1159/000489919. Epub 2018 May 14.
BACKGROUND/AIMS: This study aimed to explore whether the adoptive transfusion of autologous CD4+CD25+ regulatory T cells (CD4+CD25+ Tregs) has a therapeutic effect on Experimental autoimmune neuritis (EAN) model rats, and it provides new experimental and theoretical bases for the immunotherapy of Guillain-Barre syndrome (GBS).
CD4+CD25+ Tregs were sorted from the spleens of rats using immunomagnetic bead separation techniques combined with flow cytometry. Their in vitro inhibitory function was determined using a lymphocyte proliferation inhibition test, and their purity was confirmed by flow cytometry. Cells were stimulated using CD3/CD28 monoclonal antibodies and were cultured in culture medium containing interleukin 2 (IL-2), transforming growth factor-β (TGF-β) and rapamycin. After 15 days of amplification, CD4+CD25+ Tregs were collected and transfused into EAN model rats. Changes in the pathology and electron microscopical morphology of rat sciatic nerves in the normal group, untreated group, low-dose group (2 × 107) and high-dose group (4 × 107) were observed, and the expression of CD4+CD25+FOXP3 in peripheral blood in the four groups of rats was detected by flow cytometry.
Compared with rats in the untreated group, rats in the treatment groups had significantly reduced infiltration of inflammatory cells in the sciatic nerve, as well as myelin and axonal damage. Additionally, the CD4+CD25+ Tregs levels in peripheral blood were significantly higher than those in the untreated group (P< 0. 05). Moreover, the therapeutic effect became more significant with an increase in the dose of adoptive transfusion.
Adoptive transfusion of CD4+CD25+ Tregs into EAN model rats has significant therapeutic effects.
背景/目的：本研究旨在探讨自体CD4+CD25+调节性T细胞（CD4+CD25+ Tregs）过继性输注对实验性自身免疫性神经炎（EAN）模型大鼠是否具有治疗作用，为吉兰-巴雷综合征（GBS）的免疫治疗提供新的实验和理论依据。
采用免疫磁珠分离技术结合流式细胞术从大鼠脾脏中分选CD4+CD25+ Tregs。采用淋巴细胞增殖抑制试验检测其体外抑制功能，通过流式细胞术确认其纯度。使用CD3/CD28单克隆抗体刺激细胞，并在含有白细胞介素2（IL-2）、转化生长因子-β（TGF-β）和雷帕霉素的培养基中培养。扩增15天后，收集CD4+CD25+ Tregs并输注到EAN模型大鼠体内。观察正常组、未治疗组、低剂量组（2×107）和高剂量组（4×107）大鼠坐骨神经的病理及电镜形态变化，采用流式细胞术检测四组大鼠外周血中CD4+CD25+FOXP3的表达。
与未治疗组大鼠相比，治疗组大鼠坐骨神经中炎性细胞浸润以及髓鞘和轴突损伤明显减轻。此外，外周血中CD4+CD25+ Tregs水平显著高于未治疗组（P<0.05）。而且，随着过继性输注剂量的增加，治疗效果更显著。
将CD4+CD25+ Tregs过继性输注到EAN模型大鼠体内具有显著治疗作用。
