RNA interference screen in primary human T cells reveals FLT3 as a modulator of IL-10 levels.
RNA interference screen in primary human T cells reveals FLT3 as a modulator of IL-10 levels.
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DOI：
10.4049/jimmunol.0902443
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发表时间：
2010-01-15
期刊：
影响因子：
--
通讯作者：
Hacohen N
中科院分区：
文献类型：
--
作者：
Astier AL;Beriou G;Eisenhaure TM;Anderton SM;Hafler DA;Hacohen N
由于缺乏沉默基因功能的工具，人类原代免疫细胞的功能研究受到阻碍。在这里，我们报告了慢病毒RNAi文库在原代人T细胞中的应用。使用靶向约1，000个信号传导基因的亚基因组shRNA文库，我们鉴定了控制IL-10产生水平的新基因。IL-10是一种由几种细胞类型分泌的强效抗炎细胞因子，包括Tr 1细胞，Tr 1细胞是通过IL-10分泌发挥其抑制活性的TcB亚群。FLT 3是一种已知的造血生长因子，被发现是活化T细胞中IL-10水平的负调节因子。这是基于几个观察。首先，FLT 3及其配体FL均由T细胞活化诱导。第二，FLT 3的沉默导致IL-10水平增加，而FL的添加抑制IL-10分泌并增加FLT 3表面水平。第三，CD 46（一种已知的Tr 1调节性T细胞诱导剂）的参与上调了表面FLT 3和分泌的FL，然后抑制了T细胞中IL-10的产生。因此，FL和FLT 3形成了一个新的调节反馈环，限制了T细胞中IL-10的产生。我们的研究结果确定Flt 3作为T细胞功能的一种新的调节因子，并提供了一种从遗传学上剖析T细胞中特定通路的策略。
Functional studies of human primary immune cells have been hampered by the lack of tools to silence gene functions. Here we report the application of a lentiviral RNAi library in primary human T cells. Using a subgenomic shRNA library targeting ~1,000 signaling genes, we identified novel genes that control the levels of IL-10 produced. IL-10 is a potent anti-inflammatory cytokine secreted by several cell types, including Tr1 cells, a subset of Tregs that exert their suppressive activity through IL-10 secretion. FLT3, a known hematopoeitic growth factor, was found to be a negative regulator of IL-10 levels in activated T cells. This was based on several observations. First, FLT3 and its ligand, FL, were both induced by T cell activation. Second, silencing of FLT3 led to increased IL-10 levels while addition of FL suppressed IL-10 secretion and increased FLT3 surface levels. Third, engagement of CD46, a known inducer of Tr1 regulatory T cells, upregulated surface FLT3 and secreted FL, which then inhibited IL-10 production in T cells. Hence, FL and FLT3 form a novel regulatory feedback loop that limits IL-10 production in T cells. Our results identified Flt3 as a new regulator of T cell function and offer a strategy to genetically dissect specific pathways in T cells.
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DOI：
10.1084/jem.20052211
发表时间：
2005-12-05
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Hawrylowicz CM
通讯作者：
Hawrylowicz CM
影响因子：
7.7
作者：
Battaglia, Manuela;Stabilini, Angela;Roncarolo, Maria-Grazia
通讯作者：
Roncarolo, Maria-Grazia
影响因子：
64.5
作者：
LYMAN, SD;JAMES, L;BECKMANN, MP
通讯作者：
BECKMANN, MP
影响因子：
4.2
作者：
Braun, SE;Chen, KY;Cornetta, K
通讯作者：
Cornetta, K
影响因子：
30.5
作者：
Fitzgerald, Denise C.;Zhang, Guang-Xian;Rostami, Abdolmohamad
通讯作者：
Rostami, Abdolmohamad