Engineering of macrophages to produce IFN-γ in response to hypoxia
Engineering of macrophages to produce IFN-γ in response to hypoxia
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DOI：
10.4049/jimmunol.166.9.5374
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发表时间：
2001-05-01
影响因子：
4.4
通讯作者：
Varesio, L
中科院分区：
文献类型：
--
作者：
Carta, L;Pastorino, S;Varesio, L
小鼠巨噬细胞的激活需要来自免疫系统和环境的信号的协作。在这项研究中，我们设计了一种小鼠M Phi细胞系，使其在环境信号缺氧的唯一刺激下被激活。低氧是一种低氧压的状态，发生在几个病理组织中，与干扰素-γ协同作用，诱导完全的M phi激活。我们用低氧诱导的合成启动子(HRE3x-TK)控制的干扰素-γ基因载体(HRE3x-TK)导入ANA-1小鼠M-Phi细胞系，并对其在常氧和低氧条件下的细胞和分子生物学进行了研究。工程化M-Phi在常氧条件下表达基础水平的干扰素-伽马mRNA和蛋白，这些基因和蛋白通过将细胞转移到低氧中而显著增强。此外，它们通过诱导干扰素反应因子-1和2‘-5’-寡腺苷合成酶的表达来响应合成的干扰素-γ。在常氧条件下，构建的MPhi具有显著的La AGS和Fc受体组成水平。低氧诱导Ia和Fe的表达进一步增强。低氧诱导一氧化氮合酶表达，随后复氧导致一氧化氮的产生。总之，以诱导方式产生干扰素-γ的工程化M-Phi，作为唯一的刺激，在低氧环境下表现出新的生化和功能特性，从而避免了其他免疫系统衍生信号的共刺激需求。
Activation of murine macrophages (M phi) requires the collaboration of signals derived from the immune system and the environment. In this study, we engineered a murine M phi cell line to become activated in response to an environmental signal, hypoxia, as the sole stimulus. Hypoxia is a condition of low oxygen tension, occurring in several pathological tissues, which acts in synergy with IFN-gamma to induce full M phi activation. We transfected the ANA-1 murine M phi cell line with a construct containing the IFN-gamma gene controlled by a synthetic promoter inducible by hypoxia (HRE3x-Tk), and we characterized the cellular and molecular biology of the engineered M phi under normoxia or hypoxia. Engineered M phi in normoxia expressed basal levels of IFN-gamma mRNA and protein that were strongly augmented by shifting the cells to hypoxia. Furthermore, they responded to the synthesized IFN-gamma with induction of IFN-responsive factor-1 and 2'-5'-oligoadenylate synthase expression. Under normoxic conditions, the engineered M phi had a significant constitutive level of la Ags and Fc receptors. Hypoxia induced further augmentation of Ia and Fe expression. Finally, hypoxia induced inducible NO synthase expression, and subsequent reoxygenation led to the production of NO. In conclusion, the engineered M phi, which produce IFN-gamma in an inducible manner, express new biochemical and functional properties in response to low oxygen environment as the sole stimulus, thereby circumventing the need for costimulation by other immune system-derived signals.