Department of Veterinary Pathology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Mol Immunol. 2010 Mar;47(6):1337-46. doi: 10.1016/j.molimm.2009.12.011. Epub 2010 Jan 22.
In mammals, CpG mediated immune activation is initiated through toll-like receptor (TLR) 9 and Hsp90 via activation of MAPK/ERK and PI3K/AKT pathways. However, in the absence of TLR9 ortholog in chicken genome, the role of Hsp90 and kinase (MAPK/ERK and PI3K/AKT) pathways in initiating CpG ODN(2007) induced immune activation in chicken is not clear. Using electrophoretic mobility shift assay (EMSA) and selective inhibitors of signal transduction pathways, we determined the role of these pathways in the production of Th1 cytokines/chemokines and nitric oxide (NO) in CpG ODN(2007) treated avian macrophage cells. Hsp90alpha but not Hsp90beta is bound to CpG ODN(2007). Inhibition of Hsp90 with geldanamycin resulted in the inactivation of MAPK/ERK and PI3K/AKT pathways leading to significantly reduced levels of IFN-gamma, IL-6 and NO mRNAs in CpG ODN(2007) stimulated cells. Moreover, inhibition of ERK1/2 and PI3/AKT kinase pathways with PD985009 and LY294002, respectively, suppresses the phosphorylation of ERK2 and AKT leading to the production of decreased amounts of IFN-gamma, IL-6 and NO mRNAs in CpG ODN(2007) stimulated cells. Our results demonstrate that binding of CpG ODN(2007) to Hsp90 induces activation of ERK2 and AKT phosphorylation leading to the production of high levels of IFN-gamma, IL-6, MIP-3alpha and nitric oxide (NO). In contrast to mammals, our results suggest that Hsp90alpha but not Hsp90beta binds with the CpG ODN(2007) and may play a major role in CpG ODN(2007) induced immunoactivation in avian macrophage cells. To our knowledge, this is the first report evaluating the involvement of Hsp90 and kinase (MAPK/ERK and PI3K/AKT) pathways in CpG mediated immunostimulation in avian macrophage cells.
在哺乳动物中，CpG 介导的免疫激活是通过 Toll 样受体 (TLR) 9 和 Hsp90 启动的，这是通过激活 MAPK/ERK 和 PI3K/AKT 途径实现的。然而，在鸡基因组中缺乏 TLR9 同源物的情况下，Hsp90 和激酶 (MAPK/ERK 和 PI3K/AKT) 途径在启动 CpG ODN(2007)诱导的鸡免疫激活中的作用尚不清楚。使用电泳迁移率变动分析 (EMSA) 和信号转导途径的选择性抑制剂，我们确定了这些途径在 CpG ODN(2007)处理的禽巨噬细胞中产生 Th1 细胞因子/趋化因子和一氧化氮 (NO)中的作用。Hsp90alpha 而不是 Hsp90beta 与 CpG ODN(2007)结合。用格尔德霉素抑制 Hsp90 导致 MAPK/ERK 和 PI3K/AKT 途径失活，导致 CpG ODN(2007)刺激的细胞中 IFN-γ、IL-6 和 NO mRNA 的水平显著降低。此外，用 PD985009 和 LY294002 分别抑制 ERK1/2 和 PI3/AKT 激酶途径，抑制 ERK2 和 AKT 的磷酸化，导致 CpG ODN(2007)刺激的细胞中 IFN-γ、IL-6 和 NO mRNA 的产生量减少。我们的结果表明，CpG ODN(2007)与 Hsp90 的结合诱导 ERK2 和 AKT 磷酸化的激活，导致高水平 IFN-γ、IL-6、MIP-3alpha 和一氧化氮 (NO)的产生。与哺乳动物不同，我们的结果表明 Hsp90alpha 而不是 Hsp90beta 与 CpG ODN(2007)结合，并可能在禽巨噬细胞中 CpG ODN(2007)诱导的免疫激活中发挥主要作用。据我们所知，这是第一项评估 Hsp90 和激酶 (MAPK/ERK 和 PI3K/AKT) 途径在禽巨噬细胞中 CpG 介导的免疫刺激中的参与的报告。
