Department of Pediatrics, National Jewish Health, 1400 Jackson St., Denver, CO 80206, USA.
Am J Respir Cell Mol Biol. 2010 Apr;42(4):415-23. doi: 10.1165/rcmb.2009-0110OC. Epub 2009 Jun 5.
Candida albicans is an increasingly important pulmonary fungal pathogen. Resident alveolar macrophages are important in host defense against opportunistic fungal infections. Activation of Group IVA cytosolic phospholipase A(2)alpha (cPLA(2)alpha) in macrophages initiates arachidonic acid (AA) release for production of eicosanoids, which regulate inflammation and immune responses. We investigated the ability of C. albicans to activate cPLA(2)alpha in unprimed alveolar macrophages and after priming with granulocyte macrophage colony-stimulating factor (GM-CSF), which regulates alveolar macrophage maturation. AA was released within minutes by GM-CSF-primed but not unprimed alveolar macrophages in response to C. albicans, and was blocked by soluble glucan phosphate (S-GP). The expression of the beta-glucan receptor dectin-1 was increased in GM-CSF-primed macrophages, and AA release from GM-CSF-primed dectin-1(-/-) alveolar macrophages was reduced to basal levels. The enhanced activation of extracellular signal-regulated kinases and phosphorylation of cPLA(2)alpha on Ser-505 that occurred in GM-CSF-primed macrophages were reduced by MEK1 and Syk inhibitors, which also suppressed AA release. At later times after C. albicans infection (6 h), unprimed and GM-CSF-primed macrophages released similar levels of AA. The expression of cyclooxygenase 2 and prostanoid production at 6 hours was higher in GM-CSF-primed macrophages, but the responses were not dependent on dectin-1. However, dectin-1 contributed to the C. albicans-stimulated increase in TNF-alpha production that occurred in GM-CSF-primed macrophages. The results demonstrate that dectin-1 mediates the acute activation of cPLA(2)alpha in GM-CSF-primed alveolar macrophages, but not in the more delayed phase of AA release and GM-CSF-dependent prostanoid production.
白色念珠菌是一种日益重要的肺部真菌病原体。驻留的肺泡巨噬细胞在宿主防御机会性真菌感染方面起着重要作用。巨噬细胞中细胞溶质 IVA 磷脂酶 A(2)alpha(cPLA(2)alpha)的激活引发花生四烯酸(AA)释放,用于产生调节炎症和免疫反应的类二十烷酸。我们研究了白色念珠菌激活未成熟肺泡巨噬细胞和粒细胞巨噬细胞集落刺激因子(GM-CSF)预刺激后巨噬细胞中 cPLA(2)alpha 的能力,GM-CSF 调节肺泡巨噬细胞成熟。GM-CSF 预刺激的肺泡巨噬细胞在数分钟内释放 AA,但未预刺激的肺泡巨噬细胞则不然,而可溶性葡聚糖磷酸盐(S-GP)则阻断了 AA 的释放。GM-CSF 预刺激的巨噬细胞中β-葡聚糖受体 dectin-1 的表达增加,GM-CSF 预刺激的 dectin-1(-/-)肺泡巨噬细胞中 AA 的释放减少到基础水平。MEK1 和 Syk 抑制剂降低了 GM-CSF 预刺激的巨噬细胞中发生的细胞外信号调节激酶的激活和 cPLA(2)alpha 在 Ser-505 上的磷酸化,同时也抑制了 AA 的释放。在白色念珠菌感染后(6 小时)的后期,未预刺激和 GM-CSF 预刺激的巨噬细胞释放的 AA 水平相似。GM-CSF 预刺激的巨噬细胞中 6 小时时环氧化酶 2 和前列腺素产生的表达更高,但这些反应不依赖于 dectin-1。然而,dectin-1 有助于 GM-CSF 预刺激的巨噬细胞中白色念珠菌刺激的 TNF-α产生增加。结果表明,dectin-1 介导 GM-CSF 预刺激的肺泡巨噬细胞中 cPLA(2)alpha 的急性激活,但不介导 AA 释放和 GM-CSF 依赖性前列腺素产生的更延迟阶段。