Characterization of in vivo Dlg1 deletion on T cell development and function.
Characterization of in vivo Dlg1 deletion on T cell development and function.
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DOI：
10.1371/journal.pone.0045276
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Miceli MC
中科院分区：
文献类型：
--
作者：
Humphries LA;Shaffer MH;Sacirbegovic F;Tomassian T;McMahon KA;Humbert PO;Silva O;Round JL;Takamiya K;Huganir RL;Burkhardt JK;Russell SM;Miceli MC
T细胞膜和细胞内信号分子响应于T细胞受体（TCR）接合的极化重组已经涉及T细胞发育和效应器应答的调节。在基于siRNA的研究中，MAGUK支架蛋白和Scribble极性复合物的成员Dlg 1已被证明在T细胞极性和TCR信号特异性中发挥作用，然而Dlg 1在体内T细胞发育和功能中的作用仍不清楚。在这里，我们提出了来自三个独立衍生的dlg 1基因敲除小鼠模型的组合数据;两个种系缺陷敲除和一个条件性敲除。虽然在任何模型中均未观察到T细胞发育、TCR诱导的早期磷酸化信号传导、肌动蛋白介导的事件或增殖中的缺陷，但Jurkat T细胞中Dlg 1的急性敲低减少了IS处肌动蛋白的积累。此外，虽然Th 1型细胞因子的产生似乎不受影响的T细胞来自小鼠的dlg 1生殖缺陷，改变生产的TCR依赖性Th 1和Th 2型细胞因子中观察到的T细胞来自小鼠的条件性损失的dlg 1表达和T细胞急性Dlg 1抑制，这表明在信号转导事件中的Dlg 1活性的差异需要导致Th 1与Th 2细胞因子诱导。在这些和其他敲除模型和siRNA策略之间观察到的不一致性表明，1）替代基因的补偿性上调可能掩盖了dlg 1在dlg 1缺陷小鼠中控制TCR介导的事件中的作用，以及2）dlg 1消融开始的发育阶段可能控制补偿事件发生的程度。这些发现为使用不同dlg 1缺陷T细胞模型的各种研究中观察到的差异提供了潜在的解释，并强调了急性dlg 1消融对于避免未来Dlg 1蛋白功能研究中补偿机制上调的重要性。
The polarized reorganization of the T cell membrane and intracellular signaling molecules in response to T cell receptor (TCR) engagement has been implicated in the modulation of T cell development and effector responses. In siRNA-based studies Dlg1, a MAGUK scaffold protein and member of the Scribble polarity complex, has been shown to play a role in T cell polarity and TCR signal specificity, however the role of Dlg1 in T cell development and function in vivo remains unclear. Here we present the combined data from three independently-derived dlg1-knockout mouse models; two germline deficient knockouts and one conditional knockout. While defects were not observed in T cell development, TCR-induced early phospho-signaling, actin-mediated events, or proliferation in any of the models, the acute knockdown of Dlg1 in Jurkat T cells diminished accumulation of actin at the IS. Further, while Th1-type cytokine production appeared unaffected in T cells derived from mice with a dlg1germline-deficiency, altered production of TCR-dependent Th1 and Th2-type cytokines was observed in T cells derived from mice with a conditional loss of dlg1 expression and T cells with acute Dlg1 suppression, suggesting a differential requirement for Dlg1 activity in signaling events leading to Th1 versus Th2 cytokine induction. The observed inconsistencies between these and other knockout models and siRNA strategies suggest that 1) compensatory upregulation of alternate gene(s) may be masking a role for dlg1 in controlling TCR-mediated events in dlg1 deficient mice and 2) the developmental stage during which dlg1 ablation begins may control the degree to which compensatory events occur. These findings provide a potential explanation for the discrepancies observed in various studies using different dlg1-deficient T cell models and underscore the importance of acute dlg1 ablation to avoid the upregulation of compensatory mechanisms for future functional studies of the Dlg1 protein.
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DOI：
10.4049/jimmunol.0900973
发表时间：
2009-12-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Carrizosa E;Gomez TS;Labno CM;Klos Dehring DA;Liu X;Freedman BD;Billadeau DD;Burkhardt JK
通讯作者：
Burkhardt JK
影响因子：
32.4
作者：
Chang JT;Ciocca ML;Kinjyo I;Palanivel VR;McClurkin CE;Dejong CS;Mooney EC;Kim JS;Steinel NC;Oliaro J;Yin CC;Florea BI;Overkleeft HS;Berg LJ;Russell SM;Koretzky GA;Jordan MS;Reiner SL
通讯作者：
Reiner SL
影响因子：
4.4
作者：
Cannon, JL;Burkhardt, JK
通讯作者：
Burkhardt, JK
影响因子：
32.4
作者：
Allenspach, EJ;Cullinan, P;Sperling, AI
通讯作者：
Sperling, AI
影响因子：
32.4
作者：
Ludford-Menting, MJ;Oliaro, J;Russell, SM
通讯作者：
Russell, SM