Pechkovsky D V, Zissel G, Stamme C, Goldmann T, Ari Jaffe H, Einhaus M, Taube C, Magnussen H, Schlaak M, Müller-Quernheim J
Medical Hospital, Research Centre Borstel, Germany.
Eur Respir J. 2002 Apr;19(4):672-83. doi: 10.1183/09031936.02.00682001a.
It was hypothesized that cell-to-cell interaction between human alveolar macrophages (AM) and alveolar epithelium, might be an important factor leading to nitric oxide synthase-2 (NOS2) messenger ribonucleic acid (mRNA) and protein expression by constituent cells of the alveolar wall and/or AM. NOS2 mRNA and the protein expression patterns of human AM and alveolar epithelial cells type II (AEC-II) isolated from normal parts of lung resections of patients with pulmonary malignancies were determined. In addition, NOS2 mRNA expression in human AM co-cultured with autologous AEC-II in the presence of pro-inflammatory cytokines interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma or lipopolysaccharide (LPS) was investigated. The effect of human surfactant protein-A (SP-A) on IFN-gamma-mediated NOS2 mRNA expression in human AM was also studied. Neither NOS2 mRNA nor protein could be detected in freshly isolated, unstimulated or cytokine-stimulated AEC-II. In contrast, freshly isolated AM from bronchoalveolar lavage or lung tissue samples expressed immunoreactivity for NOS2 protein, but no NOS2 mRNA could be detected by reverse transcriptase polymerase chain reaction. All stimuli tested failed to induce NOS2 mRNA expression in human AM in vitro. Only AM-AEC-II co-culture in the presence of IFN-gamma led to NOS2 mRNA and protein expression. In situ hybridization of NOS2 mRNA on lung tissue explants and immunohistochemical staining of cytospin preparations of AM-AEC-II co-cultures demonstrated that NOS2 is expressed in AM but not in AEC-II. This co-culture effect could not be reproduced by substitution of AEC-II with SP-A. These data give evidence of a regulatory network controlling human nitric oxide synthase-2 expression in the lower respiratory tract.
据推测，人肺泡巨噬细胞（AM）与肺泡上皮细胞之间的细胞间相互作用，可能是导致肺泡壁组成细胞和/或AM中一氧化氮合酶2（NOS2）信使核糖核酸（mRNA）和蛋白表达的一个重要因素。测定了从肺部恶性肿瘤患者肺切除正常部位分离出的人AM和II型肺泡上皮细胞（AEC-II）的NOS2 mRNA及蛋白表达模式。此外，还研究了在促炎细胞因子白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α、干扰素（IFN）-γ或脂多糖（LPS）存在的情况下，人AM与自体AEC-II共培养时NOS2 mRNA的表达情况。还研究了人表面活性蛋白A（SP-A）对人AM中IFN-γ介导的NOS2 mRNA表达的影响。在新鲜分离的、未受刺激或细胞因子刺激的AEC-II中，均未检测到NOS2 mRNA和蛋白。相反，从支气管肺泡灌洗或肺组织样本中新鲜分离的AM表达了NOS2蛋白的免疫反应性，但通过逆转录聚合酶链反应未检测到NOS2 mRNA。所有测试的刺激物均未能在体外诱导人AM中NOS2 mRNA的表达。只有在IFN-γ存在的情况下，AM-AEC-II共培养才导致NOS2 mRNA和蛋白表达。肺组织外植体上NOS2 mRNA的原位杂交以及AM-AEC-II共培养的细胞涂片制剂的免疫组织化学染色表明，NOS2在AM中表达，而不在AEC-II中表达。用SP-A替代AEC-II无法重现这种共培养效应。这些数据证明了存在一个控制下呼吸道中人一氧化氮合酶2表达的调节网络。
