Alveolar macrophages and T cells from sarcoid, but not normal lung, are permissive to adenovirus infection and allow analysis of NF-κB-dependent signaling pathways
Alveolar macrophages and T cells from sarcoid, but not normal lung, are permissive to adenovirus infection and allow analysis of NF-κB-dependent signaling pathways
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DOI：
10.1165/ajrcmb.25.2.4327
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发表时间：
2001-08-01
影响因子：
6.4
通讯作者：
Foxwell, BMJ
中科院分区：
文献类型：
--
作者：
Conron, M;Bondeson, J;Foxwell, BMJ
腺病毒（Adv）介导的基因转移需要有效感染靶细胞。本研究的目的是建立是否肺泡巨噬细胞（AM）和T细胞（AT）从结节病患者允许感染腺病毒载体，如果这个属性可以用来研究细胞因子基因调控。用表达β-半乳糖苷酶或绿色荧光蛋白的Adv载体感染的结节病和正常支气管肺泡灌洗（BAL）标本分别通过荧光激活细胞分选仪（FACS）和直接免疫荧光法分析转基因表达。还使用FACS分析评估了先前与Adv感染相关的表面抗原、科萨基/腺病毒受体（CAR）、α v β 3和α v β 5整联蛋白的表达。发现结节病AM和AT有效地表达Adv转基因，不像来自正常志愿者、外周血单核细胞和外周血T细胞的AM。允许Adv感染的细胞表达CAR和α v β 5整联蛋白（对于AM也是α v β 3整联蛋白）。数据表明Adv受体的上调和感染类肉瘤AM和AT的能力与肺内的炎症环境有关。已经证明了有效的腺病毒介导的转基因递送到类肉瘤AM和AT，编码猪I κ B α的构建体。探讨肺结节病细胞因子基因表达对核因子-κ B（NF-κ B）的调控作用。类肉瘤BAL标本中I κ B α的过表达表明AM产生的肿瘤坏死因子-α和白细胞介素（IL）-6以及AT产生的干扰素（IFN）-γ依赖于NF-κ B，而AT产生的IL-4不依赖于NF-κ B。这是第一次证明在原代人T细胞内IFN-γ基因表达中需要NF-κ B。这项研究的结果对未来炎症性肺病分子通路的研究具有启示意义。
Adenovirus (Adv)-mediated gene transfer requires efficient infection of target cells. The objective of this study was to establish whether alveolar macrophages (AM) and T cells (AT) from sarcoid patients were permissive to infection with Adv vectors and if this property could be used to investigate cytokine gene regulation. Sarcoid and normal bronchoalveolar lavage (BAL) specimens infected with Adv vectors expressing either P-galactosidase or a green fluorescent protein were analyzed for transgene expression by fluorescence-activated cell sorter (FACS) and direct immunofluorescence, respectively. Expression of surface antigens previously associated with Adv infection, the coxsackie/adenovirus receptor (CAR), alphav beta3, and alphav beta5 integrins, was also assessed using FACS analysis. Sarcoid AM and AT were found to efficiently express Adv transgenes, unlike AM from normal volunteers, peripheral blood monocytes, and peripheral blood T cells. Cells permissive to Adv infection expressed the CAR and alphav beta5 integrin (also alphav beta3 integrin for AM). The data indicate that the upregulation of Adv receptors and the ability to infect sarcoid AM and AT are related to the inflammatory environment within the lung. Having demonstrated efficient Adv-mediated transgene delivery to sarcoid AM and AT, a construct encoding porcine I kappaB alpha. was then used to investigate the requirement for nuclear factor (NF)-kappaB in the regulation of cytokine gene expression in pulmonary sarcoidosis. Overexpression of I kappaB alpha in sarcoid BAL specimens indicated that tumor necrosis factor-a and interleukin (IL)-6 production by AM and interferon (IFN)-gamma production by AT is NF-kappaB dependent, whereas IL-4 production by AT is NF-kappaB independent. This is the first occasion that the requirement for NF-kappaB in IFN-gamma gene expression within primary human T cells has been demonstrated. The results of this study have implications for the future investigation of molecular pathways in inflammatory lung disease.