Assani Kaivon, Shrestha Chandra L, Robledo-Avila Frank, Rajaram Murugesan V, Partida-Sanchez Santiago, Schlesinger Larry S, Kopp Benjamin T
Center for Microbial Pathogenesis, The Research Institute at Nationwide Children's Hospital, Columbus, OH 43205.
Center for Microbial Interface Biology, Department of Microbial Infection and Immunity, The Ohio State University, Columbus, OH 43210; and.
J Immunol. 2017 Mar 1;198(5):1985-1994. doi: 10.4049/jimmunol.1502609. Epub 2017 Jan 16.
Macrophage intracellular pathogen killing is defective in cystic fibrosis (CF), despite abundant production of reactive oxygen species (ROS) in lung tissue. species can cause serious infection in CF and themselves affect key oxidase components in murine non-CF cells. However, it is unknown whether human CF macrophages have an independent defect in the oxidative burst and whether contributes to this defect in terms of assembly of the NADPH oxidase complex and subsequent ROS production. In this article, we analyze CF and non-CF human monocyte-derived macrophages (MDMs) for ROS production, NADPH assembly capacity, protein kinase C expression, and calcium release in response to PMA and CF pathogens. CF MDMs demonstrate a nearly 60% reduction in superoxide production after PMA stimulation compared with non-CF MDMs. Although CF MDMs generally have increased total NADPH component protein expression, they demonstrate decreased expression of the calcium-dependent protein kinase C conventional subclass α/β leading to reduced phosphorylation of NADPH oxidase components p47 and p40 in comparison with non-CF MDMs. Ingestion of independently contributes to and worsens the overall oxidative burst deficits in CF MDMs compared with non-CF MDMs. Together, these results provide evidence for inherent deficits in the CF macrophage oxidative burst caused by decreased phosphorylation of NADPH oxidase cytosolic components that are augmented by These findings implicate a critical role for defective macrophage oxidative responses in persistent bacterial infections in CF and create new opportunities for boosting the macrophage immune response to limit infection.
尽管肺组织中活性氧（ROS）大量产生，但巨噬细胞对细胞内病原体的杀伤在囊性纤维化（CF）中存在缺陷。这些病原体可在CF中引起严重感染，并影响小鼠非CF细胞中的关键氧化酶成分。然而，尚不清楚人类CF巨噬细胞在氧化爆发中是否存在独立缺陷，以及其是否在NADPH氧化酶复合物组装及随后的ROS产生方面导致了这一缺陷。在本文中，我们分析了CF和非CF人类单核细胞衍生巨噬细胞（MDM）对PMA和CF病原体的ROS产生、NADPH组装能力、蛋白激酶C表达及钙释放情况。与非CF MDM相比，CF MDM在PMA刺激后超氧化物产生减少了近60%。尽管CF MDM总体上NADPH成分蛋白表达增加，但与非CF MDM相比，它们的钙依赖性蛋白激酶C传统亚类α/β表达降低，导致NADPH氧化酶成分p47和p40的磷酸化减少。与非CF MDM相比，摄入这些病原体独立导致并加剧了CF MDM中整体氧化爆发缺陷。总之，这些结果为CF巨噬细胞氧化爆发中由于NADPH氧化酶胞质成分磷酸化减少而导致的固有缺陷提供了证据，而这些病原体加剧了这种缺陷。这些发现表明巨噬细胞氧化反应缺陷在CF持续性细菌感染中起关键作用，并为增强巨噬细胞免疫反应以限制感染创造了新机会。
