中华皮肤科杂志 ›› 2007, Vol. 40 ›› Issue (1): 25-27.

• 论著 • 上一篇    下一篇

Toll样受体9在SLE患者产生抗dsDNA抗体中的作用

刘慧敏1, 张瑞英2, 耿静1, 张西克1, 吴平1, 张合恩1, 姚贵申1   

  1. 1. 河北工程大学附属医院皮肤科 河北邯郸 056029;
    2. 河北工程大学附属医院血液内科
  • 收稿日期:2006-03-14 出版日期:2007-01-15 发布日期:2007-01-15

Role of TLR9 in anti-double stranded DNA antibody production in systemic lupus erythematosus

LIU Hui-min1, ZHANG Rui-yng2, GENG Jing1, ZHANG Xi-ke1, WU Ping1, ZHANG He-en1, YAO Gui-shen1   

  1. Depardnent of Dermatology, Affiliated Hospital, Hebei University of Engineering, handan 056029, China
  • Received:2006-03-14 Online:2007-01-15 Published:2007-01-15

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摘要： 目的 探讨Toll样受体(TLR9)在系统性红斑狼疮(SLE)患者抗dsDNA抗体产生中的作用.方法 密度梯度离心分离SLE患者(n=72)和正常对照者(n=30)的外周血单一核细胞(PBMC),以TLR9反义寡核昔酸转染封闭其TLR9,经天然的小牛胸腺DNA(nCTDNA)作为免疫原体外刺激后,ELISA法检测细胞培养上清液中的抗dsDNA抗体.结果 无义组、脂质体组和空白对照组均可检测到较高水平的抗dsDNA抗体(A值分别为:0.79±0.98,0.76±0.96和0.81±0.99),而反义组与其它三组相比dsDNA抗体水平明显降低,检测不到明显的抗dsDNA抗体(A值:0.06±0.04),差异均有统计学意义(P<0.001).TLR9抑制剂氯唆处理可使SLE患者PBMC抗dsDNA抗体的产生明显下降(P<0.001).结论 TLR9参与了SLE患者体内抗dsDNA抗体的产生.

关键词: 银屑病, 瘙痒, 神经生长因子, 受体, 红斑狼疮,系统性, 抗体,抗核, TLR9, 氯喳

Abstract: Objective To investigate the role and mechanism of Toll-like receptor 9(TLR-9)in anti-double stranded DNA(anti-dsDNA)antibody production in systemic lupus erythematosus(SLE). Methods Peripheral blood mononuclear cells(PBMC)were isolated from patients with SLE(n=72) and healthy control subjects(n=30)by Ficoll-Hypaque gradient centrifugation, followed by transfection with TLR9 antisense oligonucleotides to abrogate the expression of TLR-9. The PBMC were cultured in the presence with natural calf thymus DNA(nCTDNA)as immunogen, then the anti-dsDNA antibody levels in cell culture supernatants were measured by an enzyme-linked immunosorbent assay(ELISA). Results High levels of anti-dsDNA antibody could be detected in nonsense oligonucleotide (NSON) group, lipofection group, and blank control group. The absorbance value(A value)of each group was 0.79±0.98, 0.76±0.96, 0.81±0.99, respectively. Whereas, the generation of anti-dsDNA antibody was significant inhibited in antisense oligonucleuotide(ASON)group(A value=0.06±0.04, P<0.001).Furthermore, the TLR9 inhibitor chloroquine also significantly reduced the production of anti-dsDNA antibody by nCTDNA-stimulated PBMC from SLE patients(P<0.001). Conclusion These results suggest that TLR9 may play a key role in controlline anti-dsDNA antibody production in SLE.

Key words: Lupus Erythematosus, Systemic, Antibody, antinuclear, TLR9, Chloroquine