Epi-allelic Erk1 and Erk2 knockdown series for quantitative analysis of T cell Erk regulation and IL-2 production
Epi-allelic Erk1 and Erk2 knockdown series for quantitative analysis of T cell Erk regulation and IL-2 production
复制标题
DOI：
10.1016/j.molimm.2007.02.008
复制
发表时间：
2007-05-01
影响因子：
3.6
通讯作者：
Lauffenburger, Douglas A.
中科院分区：
文献类型：
--
作者：
Wille, Lucia;Kemp, Melissa L.;Lauffenburger, Douglas A.
ERK的激活通常被用来作为TCR信号通路的下游指标，通常是根据ERK1和ERK2亚型一起测量的。为了研究ERK1和ERK2调控之间的潜在差异以及TCR连接对下游的影响，我们在1136个T细胞杂交瘤中建立了一系列稳定和独立的ERK1和ERK2 shRNA敲除株。我们没有观察到相反的异构体上调的代偿作用，并发现整个上皮细胞系中总磷酸化的ERK1/2对TCR的抗CD3和多肽-MHC刺激的反应相似。此外，先前关于ERK的激活和IL-2的产生之间的线性关系的预测也得到了证实。在类似程度的ERK1/2磷酸化水平下，shRNA介导的基因敲除在减少IL-2产生方面的作用强于药物抑制MEK的作用。(C)2007爱思唯尔有限公司。保留所有权利。
Erk activation is often used as a downstream pathway indicator of TCR signaling, generally in terms of both Erk1 and Erk2 isoforms measured together. In order to investigate potential distinctions between Erk1 and Erk2 regulation and effects downstream of TCR ligation, we generated a series of stable and independent Erk1 and Erk2 shRNA knockdown lines in the 1136 T cell hybridoma. We observed no compensatory effect by opposite isoform upregulation, and found similar fractions of total phosphorylated Erk1/2 across this epi-altelic series in response to both anti-CD3 and peptide-MHC stimulation of TCR. Moreover, a previous prediction of an isoform-independent linear relationship between Erk activation and IL-2 production was confirmed. The effect of the shRNA-mediated knockdowns in reducing IL-2 production was observed to be stronger than that arising from pharmacological MEK inhibition at comparable degrees of ERK1/2 phosphorylation levels. (c) 2007 Elsevier Ltd. All rights reserved.