中华皮肤科杂志 ›› 2009, Vol. 42 ›› Issue (7): 470-472.

• 论著 • 上一篇    下一篇

SLE患者外周血单一核细胞介导T淋巴细胞对血管内皮细胞黏附的实验研究

杨骥1,储以微2,李明3   

  1. 1. 复旦大学中山医院皮肤科
    2.
    3. 上海市中山医院皮肤科
  • 收稿日期:2008-07-01 修回日期:2009-02-11 出版日期:2009-07-15 发布日期:2009-07-08
  • 通讯作者: 杨骥
  • 基金资助:

Peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus mediate the adhesion of T lymphocytes to endothelial cells

  • Received:2008-07-01 Revised:2009-02-11 Online:2009-07-15 Published:2009-07-08

摘要:

目的 探讨活动期SLE患者外周血单一核细胞(PBMC)是否介导了T淋巴细胞对血管内皮细胞的黏附。方法 体外培养人脐静脉内皮细胞(HUVEC),活动期SLE PBMC培养上清液预刺激48 h,通过RT-PCR检测PBMC培养上清液诱导HUVEC黏附分子的表达;划痕实验检测PBMC培养上清液诱导内皮细胞运动迁移能力的改变;HUVEC与T细胞共培养,研究PBMC培养上清液预刺激是否影响T细胞对HUVEC的黏附。结果 HUVEC经过活动期SLE PBMC培养上清刺激后,内皮细胞黏附分子ICAM-1 mRNA,VCAM-1 mRNA和E-钙黏蛋白mRNA表达明显增加,而IL-17抗体能明显抑制黏附分子的表达。黏附分子表达增加介导了内皮细胞运动能力增加,介导T细胞对内皮细胞的黏附,而IL-17中和抗体能抑制T细胞对内皮细胞的黏附和抑制内皮细胞的运动迁移。结论 活动期SLE PBMC培养上清液可以通过诱导血管内皮细胞黏附分子表达,黏附T细胞,介导狼疮血管炎的发生。

关键词: 红斑狼疮,系统性;活动;Th17细胞;内皮细胞;血管炎

Abstract:

Objective To observe whether peripheral blood mononuclear cells (PBMCs) from patients with active systemic lupus erythematosus (SLE) can induce the adhesion of T lymphocytes to endothelial cells. Methods Sera and PBMCs were obtained from patients with active SLE and normal human controls. PBMCs were cultivated and culture supernatants were harvested. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro with or without the presence of the sera or culture supernatants of PBMCs. Some cells were pretreated with the antibody to IL-17 before the treatment with the sera or supernatants. After another 48-hour culture, RT-PCR and real-time PCR were used to detect the mRNA expressions of adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-cadherin in HUVECs, wound healing assay to estimate the motility of HUVECs. Additionally, T lymphocytes were added to HUVECs 48 hours after stimulation with the sera or supernatants, the adhering of T lymphocytes to HUVECs was observed by microscopy. Results After stimulation with supernatants of PBMCs from patients with active SLE, the mRNA expressions of ICAM-1, VCAM-1 and E-cadherin were significantly increased in HUVEC, while the increase could be inhibited by the antibody to IL-17. The elevation of adhesion molecule expression subsequently promoted the motility of HUVEC, mediated the adhesion of T lymphocytes to HUVEC, and the antibody to IL-17 could suppress the adhesion of T lymphocytes and motility of HUVEC. Conclusion The culture supernatants of PBMCs from patients with active SLE can induce the expression of vascular cell adherin molecules and promote the adherin of T lymphocytes, which may in turn mediate the development of lupus vasculitis.