Chang Jae-Ho, Ryang Yong-Suk, Morio Tomohiro, Lee Sang-Kyou, Chang Eun-Ju
Department of Biotechnology, College of Engineering and Bioproducts Research Center, Yonsei University, Seoul 120-749, Korea.
Mol Cells. 2004 Oct 31;18(2):177-85.
Activation of NF-kappaB leads to the production of proinflammatory cytokines such as IL-12 and TNF-alpha that are involved in innate and adaptive immunity. We determined whether T. vaginalis-induced inflammatory response in macrophages associated with NF-kappaB. T. vaginalis adhesion led to transient NF-kappaB activation at 6 h but activation declined dramatically by 8 h. Super-shift assays showed that the gel-shifted complexes consisted of p65 (Rel A) and p50 (NF-kappaB1). NF-kappaB activation was accompanied by IkappaB-alpha degradation, and was inhibited by blocking T. vaginalis adhesion, indicating that the early NF-kappaB activation by T. vaginalis depends on IkappaB-alpha degradation. Quantitative real-time RT-PCR analyses revealed that the expression of TNF-alpha and IL-12 mRNA in T. vaginalis-adhesive cells was rapidly suppressed in comparison with LPS stimulation. We also observed that the parasite inhibited the nuclear translocation of NF-kappaB at 8 h, and diminished IL-12 and TNF-alpha production in response to LPS. In addition, inhibition of IkappaB-alpha degradation by MG-132 resulted in apoptosis. These results demonstrate that effects of T. vaginalis on NF-kappaB regulation are critical for cytokine production and the survival of macrophages. We suggest that there exist inhibitory mechanisms induced by T. vaginalis to evade host immunity.
核因子-κB(NF-κB)的激活会导致促炎细胞因子如白细胞介素-12(IL-12)和肿瘤坏死因子-α(TNF-α)的产生,这些细胞因子参与天然免疫和适应性免疫。我们确定了阴道毛滴虫诱导的巨噬细胞炎症反应是否与NF-κB相关。阴道毛滴虫黏附在6小时时导致NF-κB短暂激活,但在8小时时激活显著下降。超迁移分析表明,凝胶迁移复合物由p65(Rel A)和p50(NF-κB1)组成。NF-κB激活伴随着IκB-α降解,并且通过阻断阴道毛滴虫黏附而受到抑制,这表明阴道毛滴虫早期的NF-κB激活依赖于IκB-α降解。定量实时逆转录聚合酶链反应(RT-PCR)分析显示,与脂多糖(LPS)刺激相比,阴道毛滴虫黏附细胞中TNF-α和IL-12 mRNA的表达迅速受到抑制。我们还观察到,在8小时时该寄生虫抑制了NF-κB的核转位,并减少了对LPS反应时IL-12和TNF-α的产生。此外,用MG-132抑制IκB-α降解会导致细胞凋亡。这些结果表明,阴道毛滴虫对NF-κB调节的影响对于细胞因子产生和巨噬细胞存活至关重要。我们认为,阴道毛滴虫存在诱导的抑制机制以逃避宿主免疫。