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Diverse Inputs To Guide The Outcome of Mammalian Target of Rapamycin Integrates
Diverse Inputs To Guide The Outcome of Mammalian Target of Rapamycin Integrates
T
he two-signal model of TCR stimulation as Signal 1
and costimulation via CD28 and other receptors as mTOR is a large (289 kDa), highly conserved serine/threonine
Signal 2 has provided a useful paradigm for dissecting kinase initially defined as the mammalian target of the natu-
the differences in stimuli leading to T cell activation versus ral macrolide rapamycin (9). Although initially developed as
tolerance. Over the past two decades, it has become apparent an antifungal antibiotic, rapamycin is a potent immunosup-
that the outcome of Ag recognition is not merely determined pressive agent, has been employed clinically in a wide range of
by activation or tolerance; rather, there is plasticity of Th cells transplantation procedures, and has shown great promise in
such that TCR engagement can lead to a variety of different several experimental models of autoimmunity (10–12). The
CD4+ effector phenotypes, depending on the environmental exact mechanism by which rapamycin facilitates systemic
milieu (1–5). In this regard, some have referred to cytokine immunosuppression is still an area of active investigation, but
exposure as Signal 3 (6). More recently, it has become ap- the compound has been shown to influence cellular prolifer-
parent that other environmental cues such as nutrient avail- ation, differentiation, and cytokine secretion of cells belong-
ability, oxygen, growth factors, and chemokines can all make ing to both the innate and adaptive immune systems (7).
Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, The Johns mTOR, mammalian target of rapamycin; mTORC, mammalian target of rapamycin
Hopkins University School of Medicine, Baltimore, MD 21231 complex; PD-1, programmed death-1; PDK1, phosphoinositide-dependent kinase-1;
PD-L1, programmed death ligand-1; PIP3, phosphatidylinositol 3,4,5-triphosphate;
Received for publication November 28, 2011. Accepted for publication February 28,
Raptor, regulatory-associated protein of mammalian target of rapamycin; REDD, reg-
2012.
ulated in the development of DNA damage response 1; Rheb, Ras homolog enriched in
This work was supported by National Institute of Allergy and Infectious Diseases Grants brain; S1P1, sphingosine 1-phosphate receptor 1; Treg, regulatory T cell; TSC, tuberous
R01AI077610 and R01 AI091481-01. sclerosis complex.
Address correspondence and reprint requests to Dr. Jonathan D. Powell, The Johns
Hopkins University School of Medicine, 1650 Orleans Street, CRB1 Room 443, Balti- Copyright Ó 2012 by The American Association of Immunologists, Inc. 0022-1767/12/$16.00
more, MD 21231. E-mail address: poweljo@jhmi.edu
Abbreviations used in this article: AMPK, AMP-activated protein kinase; 2-DG, 2-
deoxyglucose; GSK3b, glycogen synthetase kinase-3b; KLF2, Kruppel-like factor 2;
www.jimmunol.org/cgi/doi/10.4049/jimmunol.1103143
4722 BRIEF REVIEWS: mTOR IN T CELLS
In mammalian cells, mTOR exists as one gene but forms two rum and glucocorticoid-inducible kinase 1, and protein kinase
distinct protein complexes: mTOR complex (mTORC) 1 and C (26, 27). It should be noted that Akt acts as both an up-
mTORC2, which differ in their inputs and substrates (Fig. stream regulator of mTORC1 activity (as indicated by the
1) (13). mTORC1 consists of the regulatory-associated pro- PI3K/PDK1-dependent phosphorylation at the T308 residue)
tein of mTOR (Raptor), mLST8, PRAS40, and DEPTOR. as well as a downstream target of mTORC2 (as indicated
mLST8 and DEPTOR are also found in the mTORC2 by phosphorylation at S473 residue). Akt-dependent inhibi-
complex, with the addition of RICTOR, mSIN1 proteins, tion of TSC2 (upstream of mTORC1) does not require
and PROTOR (13). Upstream of the mTORC1 complex is mTORC2 (27–29).
the small activating GTPase Ras homolog enriched in brain mTOR signaling guides CD4+ T cell fate and function. To spe-
(Rheb), the function of which is regulated by the GAP activity cifically address the potential role of mTOR in CD4+ T cell
of tuberous sclerosis complex 1 (TSC-1) and TSC-2 (14, 15). differentiation, our group selectively knocked out mTOR in
The GAP activity of TSC-1/2 can be inhibited via phosphor- T cells (30). Interestingly, CD4+ T cells lacking mTOR fail to
ylation by the kinase Akt, thereby permitting the GTP-bound differentiate into Th1, Th2, or Th17 effector cells when cul-
form of Rheb to activate mTOR (16). The activation of Akt tured in appropriate conditions in vitro. Rather, the mTOR
is facilitated by receptor-mediated activation of PI3K, which,
null T cells become Foxp3+ regulatory T cells (Tregs). The
through the production of phosphatidylinositol 3,4,5-triphos-
inability of mTOR-deficient CD4+ T cells to differentiate
phate (PIP3), activates phosphoinositide-dependent kinase-1
toward an effector phenotype is accompanied by decreased
(PDK1), which in turn activates Akt. Although the activa-
STAT4, STAT3, and STAT6 phosphorylation in response to
in T cell effector function. Selectively deletion of Rheb in lines, Pearce et al. (47) observed that when TNFR-associated
T cells specifically inhibits mTORC1 activity but maintains factor 6 was specifically deleted in T cells, CD8+ memory cell
mTORC2 activity (28). As was the case with the mTOR generation was markedly impaired. The failure of the effector
null T cells, Rheb null T cells fail to become Th1 and Th17 cells to transition into memory cells was associated with an
cells when activated under appropriate culture conditions. inability to switch to catabolism relating to fatty acid oxida-
However, somewhat surprisingly, the Rheb null T cells still tion. Based on these observations, they went on to show that
maintain the ability to differentiate into Th2 cells. Con- activating AMP-activated protein kinase (AMPK) with met-
versely, examination of T cells lacking mTORC2 activity via formin or inhibiting mTOR with rapamycin led to an in-
selective deletion of Rictor reveals that Rictor null T cells fail crease in fatty acid oxidation and a consequent increase in
to become Th2 cells in response to IL-4 but, unlike the Rheb memory generation.
null T cells, Rictor null T cells still maintain the ability to mTOR regulates T cell trafficking. The ability of naive T cells
become Th1 and Th17 cells. Another group has also condi- to circulate through secondary lymphoid tissue is facilitated
tionally deleted Rictor in T cells using a different Cre trans- by the expression of a number of cell-surface receptors,
gene and likewise observed these cells fail to become Th2 including CD62L and the chemokine receptor CCR7 (48).
cells, but interestingly, this was accompanied by a decrease in Mechanistically, the expression of CD62L, CCR7, and the
Th1 differentiation as well in this system (29). Importantly, memory marker IL-7Rb (CD127) has been linked to the
elimination of either mTORC1 or mTORC2 signaling alone FOXO family of transcription factors and Kruppel-like fac-
in T cells did not lead to the spontaneous generation of Tregs tor 2 (KLF2) (49, 50). mTORC2 activation of Akt, inhibits
TCR Activates PI3K in a dose-dependent fashion, leading to T cell activation and modulating naive 59, 61
T cell differentiation
CD28 Activates PI3K to a much greater degree than TCR stimulation alone, facilitating T cell 64, 65, 66, 67
proliferation and cytokine production
ICOS Robustly activates PI3K via cytoplasmic tail 68
OX40 Facilitates T cell memory generation and cytokine secretion by activating PI3K and AKT 70, 71, 72, 73
CTLA-4 Ligation inhibits mTOR activity via PP2a-dependent dephosphorylation of AKT; alternatively, 74, 75, 76
has been shown to directly activate PI3K
PD-1 Induces expression of the phosphatase PTEN, facilitating the degradation of PIP3 74, 77, 78, 81
IL-1 Induces mTOR activity, facilitating the development of Th17 CD4 T cells 91
IL-2 Activates PI3K, facilitating cell-cycle progression and proliferation, while inhibiting the 87
induction of T cell anergy
IL-4 Activates PI3K 88
IL-7 Regulates T cell size and metabolism downstream of PI3K/AKT/mTOR 89, 90
result in functional consequences. For example, the ability of a conserved YMNM motif and mediates Akt activation (63).
low-dose Ag to induce Foxp3+ T cells has been attributed in Ab-mediated ligation of CD28 can induce Akt activity in-
part to weak TCR-induced mTOR activity (35, 58). This is dependently of TCR stimulation (64), and constitutively ac-
particularly prominent when immature dendritic cells are tive Akt can overcome the inability of CD28-deficient cells to
used as APCs (60). Similarly, it has been shown that pre- secrete IL-2 but cannot restore their proliferative capacity
mature termination of TCR engagement promotes Foxp3+ (64).
expression due to antagonized PI3K–mTOR signaling (34). The sustained activation of PI3K and mTOR resulting from
Katzman et al. (61) have been able to correlate the duration of CD28 activation has been shown to promote proliferation in
TCR signaling with the induction of T cell activation or T cells independently of IL-2 production (65). This is the
tolerance. In their model, short-lived T cell–APC interactions consequence of optimal expression of cyclin D3 and down-
leading to tolerance are correlated with decreased mTOR regulation of the cell-cycle inhibitor p27 (66). In addition
activation. to T cell activation, CD28-mediated costimulation plays an
Although it is now clear that the Signal 2 is in reality com- important role in enhancing glycolysis and glucose uptake
prised of multiple ligand receptor interactions, perhaps the (67). This process has been shown to be dependent on PI3K/
best-described costimulatory signal on T cells is the interaction Akt signaling and involves the rapid upregulation in expres-
between CD28 and its two known ligands B7.1 and B7.2. sion of the Glut1 glucose transporter (67).
CD28 facilitates the nuclear translocation of NF-kB and en- The costimulatory signal provided by CD28 ligation on
hances transcription and translation of IL-2 (62). Thus, one naive T cells is important for the initiation of a T cell response,
means CD28 ligation can promote mTOR activity is in an but additional receptor–ligand interactions can also provide
autocrine fashion through IL-2 signaling. The ligation of a costimulatory signal and fine-tune the T cell activation
CD28 on an activated T cell can also directly activate PI3K. profile at the time of initial activation. The ICOS/ICOS li-
PI3K binds the phosphorylated cytoplasmic tail of CD28 at gand interaction is a potent inducer of PI3K activation. In
The Journal of Immunology 4725
fact, studies suggest that the direct binding of PI3K to the is mediated through the downregulation of the Akt–mTOR
conserved YMFM motif on the ICOS cytoplasmic tail leads axis signaling.
to more robust activation than that induced by CD28 en- Cytokines/IFNs/chemokines. mTOR signaling plays a role in
gagement (68). Detailed studies examining the role of ICOS regulating the downstream consequences of a number of
on regulating mTOR activity in T cells have yet to be per- immunologically relevant cytokines. Early studies identified
formed. However, given the prominent role that ICOS plays mTOR activity as being increased upon IL-2–induced stim-
in PI3K activation in T cells, one would predict that ICOS ulation (83). IL-2–induced mTOR activation was shown to
will also play an important role in regulating mTOR. be important for facilitating cell-cycle progression and prolif-
The surface receptor OX40 (CD134) has recently gained eration (83). These observations led to a series of studies
recognition as a potent costimulatory molecule that comple- examining the ability of mTOR to regulate T cell anergy
ments the activity of CD28 and ICOS. A member of the (65, 84–86). It has been shown that the ability of IL-2 to
TNF-a receptor superfamily, OX40 expression is strongly, both prevent and reverse T cell anergy is dependent upon
though transiently, induced following TCR stimulation in mTOR activation (85, 87). Other common g-chain cytokine
both CD4 and CD8 T cells, peaking in expression 48 h after receptors also activate mTOR. Like IL-2, IL-4R signaling is
stimulation and returning to baseline by 120 h (69). Ligation another potent inducer of T cell proliferation but has the
of this receptor, either through interaction with its APC- added ability to skew naive CD4 T cell to a Th2 phenotype.
restricted ligand OX40L (CD252) or by Ab-mediated cross- The cytoplasmic tail of the IL-4R possesses five evolutionarily
linking, facilitates increased clonal proliferation, cell survival, conserved tyrosine residues that have been shown to differ-
stimulation (99). For activated Th1/Th2 CD4 T cells, TSC2 by GSK-3b is dependent on prior phosphorylation of
mTOR activity is required for CCR5/CCL5 (RANTES)- the substrate by AMPK at the S1345 residue (114). The in-
mediated migration that is dependent upon 4EBP-mediated teraction of Wnt with its receptor on the plasma membrane of
translation (100). However, not all chemokines depend on most mammalian cells inhibits the activity of GSK-3b. As such,
mTOR activation. For example, mTOR signaling is dis- Wnt signaling can promote mTORC1 activity.
pensable for CCL19 (Mip3b)-mediated migration (99). Low oxygen tension (as might be experienced in a tumor
Although best known for regulating appetite and energy microenvironment) can also regulate mTOR activity. It has
expenditure, the adipokine leptin also plays a significant role been shown that in the setting of low oxygen, the hypoxia-
in regulating the functionality and proliferative capacity of induced factor protein regulated in the development of
T cells through its ability to stimulate mTOR activity (101– DNA damage response 1 (REDD1) can inhibit mTOR by
103). In the absence of leptin receptor stimulation, autore- promoting the assembly and activation of TSC (115). Cells
active CD4+ T cells exhibit decreased expression of the anti- lacking REDD1 show continued mTOR activity even under
apoptotic factor Bcl2, an impaired ability to skew to a Th1/ conditions of nutrient withdrawal (116), whereas hypoxia
Th17 phenotype, and a failure to upregulate mTOR activity facilitates the REDD1-mediated activation of the TSC1/2 by
(103). Further, leptin acting via the mTOR signaling pathway facilitating the stabilization of TSC1/2 by 14-3-3 protein
has been shown to provide a link between energy status and (117). Although hypoxia is a potent regulator of mTOR ac-
Treg function (104). tivity, mTORC-1 regulates the expression of the canonical
The lysophospholipid S1P is another potent inducer of hypoxia response element hypoxia inducible factor-1 (118,
57. Badou, A., M. Savignac, M. Moreau, C. Leclerc, G. Foucras, G. Cassar, P. Paulet, 83. Abraham, R. T., and G. J. Wiederrecht. 1996. Immunopharmacology of rapa-
D. Lagrange, P. Druet, J. C. Guéry, and L. Pelletier. 2001. Weak TCR stimulation mycin. Annu. Rev. Immunol. 14: 483–510.
induces a calcium signal that triggers IL-4 synthesis, stronger TCR stimulation 84. Allen, A., Y. Zheng, L. Gardner, M. Safford, M. R. Horton, and J. D. Powell.
induces MAP kinases that control IFN-gamma production. Eur. J. Immunol. 31: 2004. The novel cyclophilin binding compound, sanglifehrin A, disassociates G1
2487–2496. cell cycle arrest from tolerance induction. J. Immunol. 172: 4797–4803.
58. Gottschalk, R. A., E. Corse, and J. P. Allison. 2010. TCR ligand density and 85. Powell, J. D., C. G. Lerner, and R. H. Schwartz. 1999. Inhibition of cell cycle
affinity determine peripheral induction of Foxp3 in vivo. J. Exp. Med. 207: 1701– progression by rapamycin induces T cell clonal anergy even in the presence of
1711. costimulation. J. Immunol. 162: 2775–2784.
59. Exley, M., L. Varticovski, M. Peter, J. Sancho, and C. Terhorst. 1994. Association 86. Vanasek, T. L., A. Khoruts, T. Zell, and D. L. Mueller. 2001. Antagonistic roles
of phosphatidylinositol 3-kinase with a specific sequence of the T cell receptor zeta for CTLA-4 and the mammalian target of rapamycin in the regulation of clonal
chain is dependent on T cell activation. J. Biol. Chem. 269: 15140–15146. anergy: enhanced cell cycle progression promotes recall antigen responsiveness. J.
60. Jonuleit, H., E. Schmitt, G. Schuler, J. Knop, and A. H. Enk. 2000. Induction of Immunol. 167: 5636–5644.
interleukin 10-producing, nonproliferating CD4(+) T cells with regulatory prop- 87. Duré, M., and F. Macian. 2009. IL-2 signaling prevents T cell anergy by inhibiting
erties by repetitive stimulation with allogeneic immature human dendritic cells. J. the expression of anergy-inducing genes. Mol. Immunol. 46: 999–1006.
Exp. Med. 192: 1213–1222. 88. Stephenson, L. M., D. S. Park, A. L. Mora, S. Goenka, and M. Boothby. 2005.
61. Katzman, S. D., W. E. O’Gorman, A. V. Villarino, E. Gallo, R. S. Friedman, Sequence motifs in IL-4R alpha mediating cell-cycle progression of primary
M. F. Krummel, G. P. Nolan, and A. K. Abbas. 2010. Duration of antigen re- lymphocytes. J. Immunol. 175: 5178–5185.
ceptor signaling determines T-cell tolerance or activation. Proc. Natl. Acad. Sci. 89. Barata, J. T., A. Silva, J. G. Brandao, L. M. Nadler, A. A. Cardoso, and
USA 107: 18085–18090. V. A. Boussiotis. 2004. Activation of PI3K is indispensable for interleukin 7-
62. Verweij, C. L., M. Geerts, and L. A. Aarden. 1991. Activation of interleukin-2 mediated viability, proliferation, glucose use, and growth of T cell acute lym-
gene transcription via the T-cell surface molecule CD28 is mediated through an phoblastic leukemia cells. J. Exp. Med. 200: 659–669.
NF-kB-like response element. J. Biol. Chem. 266: 14179–14182. 90. Rathmell, J. C., E. A. Farkash, W. Gao, and C. B. Thompson. 2001. IL-7 en-
63. Harada, Y., M. Tokushima, Y. Matsumoto, S. Ogawa, M. Otsuka, K. Hayashi, hances the survival and maintains the size of naive T cells. J. Immunol. 167: 6869–
B. D. Weiss, C. H. June, and R. Abe. 2001. Critical requirement for the 6876.
membrane-proximal cytosolic tyrosine residue for CD28-mediated costimulation 91. Gulen, M. F., Z. Kang, K. Bulek, W. Youzhong, T. W. Kim, Y. Chen,
in vivo. J. Immunol. 166: 3797–3803. C. Z. Altuntas, K. Sass Bak-Jensen, M. J. McGeachy, J. S. Do, et al. 2010. The
GSK-3 phosphorylation and NF-AT activation in Jurkat T cells. Biochem. Biophys. 118. Hudson, C. C., M. Liu, G. G. Chiang, D. M. Otterness, D. C. Loomis, F. Kaper,
Res. Commun. 332: 339–346. A. J. Giaccia, and R. T. Abraham. 2002. Regulation of hypoxia-inducible factor
111. Nath, N., S. Giri, R. Prasad, M. L. Salem, A. K. Singh, and I. Singh. 2005. 5- 1alpha expression and function by the mammalian target of rapamycin. Mol. Cell.
aminoimidazole-4-carboxamide ribonucleoside: a novel immunomodulator with Biol. 22: 7004–7014.
therapeutic efficacy in experimental autoimmune encephalomyelitis. J. Immunol. 119. Nakamura, H., Y. Makino, K. Okamoto, L. Poellinger, K. Ohnuma, C. Morimoto,
175: 566–574. and H. Tanaka. 2005. TCR engagement increases hypoxia-inducible factor-1
112. Jiang, W., Z. Zhu, and H. J. Thompson. 2008. Modulation of the activities of alpha protein synthesis via rapamycin-sensitive pathway under hypoxic con-
AMP-activated protein kinase, protein kinase B, and mammalian target of rapamycin ditions in human peripheral T cells. J. Immunol. 174: 7592–7599.
by limiting energy availability with 2-deoxyglucose. Mol. Carcinog. 47: 616–628. 120. Dang, E. V., J. Barbi, H. Y. Yang, D. Jinasena, H. Yu, Y. Zheng, Z. Bordman,
113. Cham, C. M., and T. F. Gajewski. 2005. Glucose availability regulates IFN- J. Fu, Y. Kim, H. R. Yen, et al. 2011. Control of T(H)17/T(reg) balance by
gamma production and p70S6 kinase activation in CD8+ effector T cells. J. hypoxia-inducible factor 1. Cell 146: 772–784.
Immunol. 174: 4670–4677. 121. Shi, L. Z., R. Wang, G. Huang, P. Vogel, G. Neale, D. R. Green, and H. Chi.
114. Inoki, K., H. Ouyang, T. Zhu, C. Lindvall, Y. Wang, X. Zhang, Q. Yang, 2011. HIF1alpha-dependent glycolytic pathway orchestrates a metabolic check-
C. Bennett, Y. Harada, K. Stankunas, et al. 2006. TSC2 integrates Wnt and energy point for the differentiation of TH17 and Treg cells. J. Exp. Med. 208: 1367–
signals via a coordinated phosphorylation by AMPK and GSK3 to regulate cell 1376.
growth. Cell 126: 955–968. 122. Sancak, Y., T. R. Peterson, Y. D. Shaul, R. A. Lindquist, C. C. Thoreen, L. Bar-
115. Brugarolas, J., K. Lei, R. L. Hurley, B. D. Manning, J. H. Reiling, E. Hafen, Peled, and D. M. Sabatini. 2008. The Rag GTPases bind raptor and mediate
L. A. Witters, L. W. Ellisen, and W. G. Kaelin, Jr. 2004. Regulation of mTOR amino acid signaling to mTORC1. Science 320: 1496–1501.
function in response to hypoxia by REDD1 and the TSC1/TSC2 tumor sup- 123. Cobbold, S. P., E. Adams, C. A. Farquhar, K. F. Nolan, D. Howie, K. O. Lui,
pressor complex. Genes Dev. 18: 2893–2904. P. J. Fairchild, A. L. Mellor, D. Ron, and H. Waldmann. 2009. Infectious tol-
116. Sofer, A., K. Lei, C. M. Johannessen, and L. W. Ellisen. 2005. Regulation of erance via the consumption of essential amino acids and mTOR signaling. Proc.
mTOR and cell growth in response to energy stress by REDD1. Mol. Cell. Biol. Natl. Acad. Sci. USA 106: 12055–12060.
25: 5834–5845. 124. Hidayat, S., K. Yoshino, C. Tokunaga, K. Hara, M. Matsuo, and K. Yonezawa.
117. DeYoung, M. P., P. Horak, A. Sofer, D. Sgroi, and L. W. Ellisen. 2008. Hypoxia 2003. Inhibition of amino acid-mTOR signaling by a leucine derivative
regulates TSC1/2-mTOR signaling and tumor suppression through REDD1- induces G1 arrest in Jurkat cells. Biochem. Biophys. Res. Commun. 301: 417–
mediated 14-3-3 shuttling. Genes Dev. 22: 239–251. 423.