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- Characterization of in vivo Dlg1 deletion on T cell development and function
- Humphries, Lisa A.; Shaffer, Meredith H.; Sacirbegovic, Faruk; Tomassian, Tamar; McMahon, Kerrie-Ann; Humbert, Patrick O.; Silva, Oscar; Round, June L.; Takamiya, Kogo; Huganir, Richard L.; Burkhardt, Janis K.; Russell, Sarah M.; Miceli, M. Carrie
- Background: 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. Methodology/Principal Findings: 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. Conclusions/Significance: 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.
- Publication type
- Journal article
- Research centre
- Swinburne University of Technology. Faculty of Engineering and Industrial Sciences. Centre for Micro-Photonics
- PLoS ONE, Vol. 7, no. 9 (Sep 2012), article no. e45276
- Publication year
- Cell development; Cell functions; Conditional knockouts; Discs large proteins; Dlg1; Dlg1 deficiency; Germline deficient knockouts; In vivo; Mice; T cells
- Public Library of Science
- Publisher URL
- Copyright © 2012 Humphries et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.5/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The published version is reproduced in accordance with this policy.
- Additional information
- The authors acknowledge support from the National Institutes of Health (http://www.nih.gov) grant P01 CA093615, the National Institutes of Health training grant T32-HD07516, the National Institutes of Health grant R01-AI067253-10, a UCLA (University of California, Los Angeles) dissertation year fellowship, a UCLA (University of California, Los Angeles) Microbial Pathogenesis Training Grant 2-T32-AI-07323, an Arthritis Foundation Postdoctoral Fellowship, the Australian National Health and Medical Research Council and the Australian Research Council.
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