McGill Centre for the Study of Host Resistance, The Research Institute of the McGill University Health Centre, Montreal, Quebec, Canada.
PLoS Negl Trop Dis. 2013;7(1):e2015. doi: 10.1371/journal.pntd.0002015. Epub 2013 Jan 17.
Leprosy is a persistent infectious disease caused by Mycobacterium leprae that still affects over 200,000 new patients annually. The host genetic background is an important risk factor for leprosy susceptibility and the PARK2 gene is a replicated leprosy susceptibility candidate gene. The protein product of PARK2, Parkin, is an E3 ubiquitin ligase that is involved in the development of various forms of Parkinsonism. The human macrophage is both a natural host cell of M. leprae as well as a primary mediator of natural immune defenses, in part by secreting important pro-inflammatory cytokines and chemokines. Here, we report that down-regulation of Parkin in THP-1 macrophages, human monocyte-derived macrophages and human Schwann cells resulted in a consistent and specific decrease in interleukin-6 (IL-6) and monocyte chemoattractant protein 1 (MCP-1/CCL2) production in response to mycobacteria or LPS. Interestingly, production of IL-6 at 6 hours by THP-1 cells stimulated with live M. leprae and M. bovis BCG was dependent on pretreatment with 1,25-dihydroxyvitamin D(3) (VD). Parkin knockdown in VD-treated cells blocked IL-6 induction by mycobacteria. However, IκB-α phosphorylation and levels of IκB-ξ, a nuclear protein required for IL-6 expression, were not affected by Parkin silencing. Phosphorylation of MAPK ERK1/2 and p38 was unaffected by Parkin silencing while JNK activation was promoted but did not explain the altered cytokine production. In a final set of experiments we found that genetic risk factors of leprosy located in the PARK2 promoter region were significantly correlated with M. leprae sonicate triggered CCL2 and IL6 transcript levels in whole blood assays. These results associated genetically controlled changes in the production of MCP-1/CCL2 and IL-6 with known leprosy susceptibility factors.
麻风病是一种由麻风分枝杆菌引起的慢性传染病,每年仍有超过 20 万新发病例。宿主遗传背景是麻风病易感性的重要危险因素,PARK2 基因是一个已被复制的麻风病易感性候选基因。PARK2 的蛋白产物 Parkin 是一种 E3 泛素连接酶,参与各种形式帕金森病的发生。人类巨噬细胞既是麻风分枝杆菌的天然宿主细胞,也是天然免疫防御的主要介质,部分原因是它分泌重要的促炎细胞因子和趋化因子。在这里,我们报告 Parkin 在 THP-1 巨噬细胞、人单核细胞来源的巨噬细胞和人施万细胞中的下调导致对分枝杆菌或 LPS 反应时白细胞介素 6(IL-6)和单核细胞趋化蛋白 1(MCP-1/CCL2)的产生持续和特异性下降。有趣的是,用活的麻风分枝杆菌和牛分枝杆菌 BCG 刺激的 THP-1 细胞在 6 小时产生的 IL-6 依赖于用 1,25-二羟维生素 D(VD)预处理。在 VD 处理的细胞中敲低 Parkin 可阻断分枝杆菌诱导的 IL-6 诱导。然而,Parkin 沉默并不影响 IκB-α 磷酸化和核蛋白 IκB-ξ 的水平,后者是 IL-6 表达所必需的。MAPK ERK1/2 和 p38 的磷酸化不受 Parkin 沉默的影响,而 JNK 激活被促进,但不能解释改变的细胞因子产生。在最后一组实验中,我们发现位于 PARK2 启动子区域的麻风病遗传风险因素与麻风分枝杆菌超声裂解物触发的全血测定中 CCL2 和 IL6 转录物水平显著相关。这些结果将 MCP-1/CCL2 和 IL-6 的遗传控制变化与已知的麻风病易感性因素联系起来。