系统性红斑狼疮患者外周血中调节性T细胞相关分子TGF-β表达缺陷的研究

发布时间：2018-05-29 14:46

本文选题：红斑狼疮 + 调节性T细胞　；参考：《南京医科大学学报(自然科学版)》2016年11期

【摘要】：目的 :研究与系统性红斑狼疮(systemic lupus erythematosus,SLE)患者调节性T细胞(regulatory T cell,Treg)功能密切相关的分子转化生长因子(transforming growth factor,TGF-β)的转录、表达、分泌水平的异常。方法:分离正常人和SLE患者的外周血单个核细胞(peripheral blood mononuclear cell,PBMC)和CD4+CD25+T细胞,以抗-CD3和抗-CD28刺激培养48 h,获取细胞,以三色流式细胞术检测CD4+CD25+LAP+T及CD8+CD25+LAP+T细胞,分析各亚群细胞比值;提取总m RNA,q RT-PCR测定TGF-β1的表达;收取上清用ELISA法检测总TGF-β1和游离的活化TGF-β1水平。结果 :活动性SLE患者CD4+T细胞中LAP+/CD4+T、CD25+LAP+/CD4+T、LAP+/CD4+CD25+比值较正常人显著升高,抗-CD3、抗-CD28抗体刺激后LAP+/CD4+T、CD25+LAP+/CD4+T、LAP+/CD4+CD25+比值进一步升高,与正常人比较差异有统计学意义(P0.05);但均与疾病活动度无显著相关性;无论刺激与未刺激,SLE患者CD4+CD25+LAP+细胞中LAP染色的荧光强度亦高于相应正常组;而抗-CD3、抗-CD28抗体刺激后SLE患者PBMC及CD4+CD25+T细胞培养上清中总TGF-β1和游离的活化TGF-β1水平均显著低于正常人(P0.05);q RT-PCR证实,受刺激后CD4+CD25+T细胞内TGF-β1的m RNA转录水平低于正常人。结论:尽管活动性SLE患者CD4+CD25+T细胞膜表面潜伏态TGF-β(LAP)的表达较正常人明显增高,但其TGF-β表达依然存在缺陷,主要表现为TGF-β1的转录、分泌水平降低,游离的活化TGF-β1产生障碍,从而可能削弱了Treg细胞的免疫抑制作用。新鲜分离的以及受刺激后的CD4+CD25+T细胞LAP表达增高可能反映了活动性SLE患者T细胞亚群的活化和CD4+CD25+LAP+Treg细胞的反应性扩增。
[Abstract]:Objective: to study the transcription, expression and secretion of transforming growth factor- 尾 (TGF- 尾) in regulatory T cells of patients with systemic lupus erythematosus (lupus erythematosus). Methods: peripheral blood mononuclear cells (PBMCs) and CD4 CD25 T cells were isolated from normal subjects and patients with SLE. The cells were cultured for 48 h after stimulation with anti-CD3 and anti-CD28. The CD4 CD25 LAP T and CD8 CD25 LAP T cells were detected by tricolor flow cytometry. The expression of TGF- 尾 1 was determined by total mRNAq RT-PCR, and the levels of total TGF- 尾 1 and free activated TGF- 尾 1 were detected by ELISA method in the supernatant. Results: the ratio of LAP / CD4 / CD25 LAP / CD4 TLAP / CD4 CD25 in CD4 T cells of active SLE patients was significantly higher than that of normal controls. The ratio of LAP / CD4 T T T CD 25 LAP / CD 4 TLAP / CD 4 CD25 was further increased after the stimulation of anti CD 3 and anti CD CD 28 antibodies. There was a significant difference between CD4 CD25 LAP cells and normal controls (P 0.05), but there was no significant correlation between LAP staining and disease activity. The fluorescence intensity of LAP staining in CD4 CD25 LAP cells of patients with or without stimulation was higher than that of normal controls. However, the levels of total TGF- 尾 1 and free activated TGF- 尾 1 in the supernatants of PBMC and CD4 CD25 T cells stimulated by anti-CD3 and anti-CD28 antibodies were significantly lower than those of normal controls (P 0.05 / Q RT-PCR). The m RNA transcription level of TGF- 尾 1 in CD4 CD25 T cells after stimulation was lower than that in normal controls. Conclusion: although the expression of latent TGF- 尾 LAPs on the surface of CD4 CD25 T cell membrane in active SLE patients is significantly higher than that in normal controls, the expression of TGF- 尾 is still defective, mainly due to the decrease of TGF- 尾 1 transcription and secretion, and the disturbance of free activation of TGF- 尾 1. This may weaken the immunosuppressive effect of Treg cells. The increased expression of LAP in freshly isolated and stimulated CD4 CD25 T cells may reflect the activation of T cell subsets and the reactive expansion of CD4 CD25 LAP Treg cells in active SLE patients.
【作者单位】：南京医科大学免疫学系;南京市红十字血液中心;
【分类号】：R593.241

【相似文献】