Abstract
The mammalian Target of Rapamycin (mTOR) defines a crucial link between nutrient sensing and immune function. In CD4+ T cells, mTOR has been shown to play a critical role in regulating effector and regulatory T cell differentiation as well as the decision between full activation versus the induction of anergy. In this chapter, we describe how our group has employed the Cre-lox technology to genetically delete components of the mTOR signaling complex in T cells. This has enabled us to specifically interrogate mTOR function in T cells both in vitro and in vivo. We also describe techniques used to assay immune function and signaling in mTOR-deficient T cells at the single-cell level.
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References
Sabatini, D. M. (2006) mTOR and cancer: insights into a complex relationship, Nat Rev Cancer 6, 729–734.
Delgoffe, G. M., Kole, T. P., Zheng, Y., Zarek, P. E., Matthews, K. L., Xiao, B., Worley, P. F., Kozma, S. C., and Powell, J. D. (2009) The mTOR kinase differentially regulates effector and regulatory T cell lineage commitment, Immunity 30, 832–844.
Araki, K., Turner, A. P., Shaffer, V. O., Gangappa, S., Keller, S. A., Bachmann, M. F., Larsen, C. P., and Ahmed, R. (2009) mTOR regulates memory CD8 T-cell differentiation, Nature 460, 108–112.
Rao, R. R., Li, Q., Odunsi, K., and Shrikant, P. A. The mTOR kinase determines effector versus memory CD8+ T cell fate by regulating the expression of transcription factors T-bet and Eomesodermin, Immunity 32, 67–78.
Kopf, H., de la Rosa, G. M., Howard, O. M., and Chen, X. (2007) Rapamycin inhibits differentiation of Th17 cells and promotes generation of FoxP3+ T regulatory cells, Int Immuno-pharmacol 7, 1819–1824.
Valmori, D., Tosello, V., Souleimanian, N. E., Godefroy, E., Scotto, L., Wang, Y., and Ayyoub, M. (2006) Rapamycin-mediated enrichment of T cells with regulatory activity in stimulated CD4+ T cell cultures is not due to the selective expansion of naturally occurring regulatory T cells but to the induction of regulatory functions in conventional CD4+ T cells, J Immunol 177, 944–949.
Battaglia, M., Stabilini, A., and Roncarolo, M. G. (2005) Rapamycin selectively expands CD4  +  CD25  +  FoxP3+ regulatory T cells, Blood 105, 4743–4748.
Wang, B., Xiao, Z., Chen, B., Han, J., Gao, Y., Zhang, J., Zhao, W., Wang, X., and Dai, J. (2008) Nogo-66 promotes the differentiation of neural progenitors into astroglial lineage cells through mTOR-STAT3 pathway, PLoS One 3, e1856.
Guertin, D. A., and Sabatini, D. M. (2007) Defining the role of mTOR in cancer, Cancer Cell 12, 9–22.
Zheng, Y., Delgoffe, G. M., Meyer, C. F., Chan, W., and Powell, J. D. (2009) Anergic T cells are metabolically anergic, J Immunol 183, 6095–6101.
Guertin, D. A., and Sabatini, D. M. (2009) The pharmacology of mTOR inhibition, Sci Signal 2, pe24.
Sarbassov, D. D., Ali, S. M., Sengupta, S., Sheen, J. H., Hsu, P. P., Bagley, A. F., Markhard, A. L., and Sabatini, D. M. (2006) Prolonged rapamycin treatment inhibits mTORC2 assembly and Akt/PKB, Mol Cell 22, 159–168.
Delgoffe, G. M., and Powell, J. D. (2009) mTOR: taking cues from the immune microenvironment, Immunology 127, 459–465.
Acknowledgments
The authors would like to thank members of the Powell laboratory for technical assistance in optimizing these models. In addition, we would like to thank Dr. Sara C. Kozma (U. Cincinnati) for generating the original floxed mouse lines. This work was supported by R01AI077610-01A2.
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Delgoffe, G.M., Powell, J.D. (2012). Exploring Functional In Vivo Consequences of the Selective Genetic Ablation of mTOR Signaling in T Helper Lymphocytes. In: Weichhart, T. (eds) mTOR. Methods in Molecular Biology, vol 821. Humana Press. https://doi.org/10.1007/978-1-61779-430-8_20
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DOI: https://doi.org/10.1007/978-1-61779-430-8_20
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