在未經治療活動性全身性紅斑性狼瘡病患以CpG去氧核糖寡核甘酸連結類鐸蛋白受體-9會減弱腫瘤壞死因子-α的產生

CpG ODN ligation of TLR-9 attenuate TNF-α production in untreated active SLE patients

目的：本研究探討在未經過治療且處於疾病活動期的全身性紅斑性狼瘡患者，與性別、年齡對應的健康受試者以CpG去氧核糖寡核甘酸連結類鐸蛋白受體9對於腫瘤壞死因子-α（TNF-α）的產生，是否有差異。材料與方法：13位全身性紅斑性狼瘡患者與8位健康受試者的周邊血液單核球，以CpG去氧核糖寡核甘酸和phorbol myristate acetate及 ionomycin刺激，分別以反轉錄聚合酶反應和酵素免疫法分析TNF-α的mRNA表達量和蛋白的產量。結果：全身性紅斑性狼瘡患者和健康受試者，以CpG去氧核糖寡核甘酸連結類鐸蛋白受體-9，都會增加週邊血液單核球產生TNF-α，但是在全身性紅斑性狼瘡患者產生的TNF-α的量比健康受試者低，並且達統計上顯著差異，對週邊血液單核球TNF-α核糖核酸的表達量在這兩組則未達顯著差異。結論：未經過治療且處於疾病活動期的全身性紅斑性狼瘡患者，其周邊血液單核球在類鐸蛋白受體-9被CpG去氧核糖寡核甘酸連結後減弱了TNF-α的產生，此一結果反應出這個重要的前驅發炎激素在全身性紅斑性狼瘡患者失去正常調節的現象，而這個現象可能和全身性紅斑性狼瘡患者感染到帶有CpG序列的細菌或病毒時其內生性免疫力會減弱有關。

Objective: In the present study, we determined the immunomodulatory effect of toll-like receptor-9 (TLR-9) ligation with hypomethylated CpG oligodeoxynucleotides (CpG ODNs) on TNF-α release in untreated systemic lupus erythematosus (SLE) patients and in a cohort of sex- and age-matched healthy individuals. Methods: Peripheral blood mononuclear cells (PBMCs) from 13 SLE patients and 8 healthy individuals were stimulated with CpG-ODNs in the presence of phorbol myristate acetate and ionomycin. RT-PCR and ELISA assays were used for analysis of TNF-α mRNA expression and protein secretion. Results: CpG ODNs increased TLR9-mediated TNF-α protein release in supernatant of PBMC culture in SLE and healthy individuals although there were no significant differences of TNF-α mRNA expression between these two groups. The level of TNF-α protein production was significantly lower in SLE patients than in normal controls (592.5 ± 202.8 ng/mL vs. 1369.8 ± 277.3 ng/mL, p =0.033). Conclusion: The lower TNF-α production by PBMCs after TLR-9 ligation in untreated SLE patients reflected the dysregulation of this important proinflammatory cytokine. This dysregulation may be involved in the pathogenesis of attenuated innate immunity when patients are exposed to bacteria or viruses with CpG-rich DNA.