The susceptibility of CD4 T cells to human immunodeficiency virus 1 (HIV-1) infection is regulated by glucose and glutamine metabolism, but the relative contributions of these nutrients to infection are not known. Here we show that glutaminolysis is the major pathway fuelling the tricarboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS) in T-cell receptor-stimulated naïve, as well as memory CD4, subsets and is required for optimal HIV-1 infection. Under conditions of attenuated glutaminolysis, the α-ketoglutarate (α-KG) TCA rescues early steps in infection; exogenous α-KG promotes HIV-1 reverse transcription, rendering both naïve and memory cells more sensitive to infection. Blocking the glycolytic flux of pyruvate to lactate results in altered glucose carbon allocation to TCA and pentose phosphate pathway intermediates, an increase in OXPHOS and augmented HIV-1 reverse transcription. Moreover, HIV-1 infection is significantly higher in CD4 T cells selected on the basis of high mitochondrial biomass and OXPHOS activity. Therefore, the OXPHOS/aerobic glycolysis balance is a major regulator of HIV-1 infection in CD4 T lymphocytes.
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The data that support the findings of this study are available from the corresponding authors upon request.
Zack, J. A., Kim, S. G. & Vatakis, D. N. HIV restriction in quiescent CD4+ T cells. Retrovirology 10, 37 (2013).
Rathmell, J. C., Elstrom, R. L., Cinalli, R. M. & Thompson, C. B. Activated Akt promotes increased resting T cell size, CD28-independent T cell growth, and development of autoimmunity and lymphoma. Eur. J. Immunol. 33, 2223–2232 (2003).
Frauwirth, K. A. et al. The CD28 signaling pathway regulates glucose metabolism. Immunity 16, 769–777 (2002).
Manel, N. et al. The HTLV receptor is an early T-cell activation marker whose expression requires de novo protein synthesis. Blood 101, 1913–1918 (2003).
Curi, R. et al. Glutamine, gene expression, and cell function. Front. Biosci. 12, 344–357 (2007).
Fuchs, B. C., Finger, R. E., Onan, M. C. & Bode, B. P. ASCT2 silencing regulates mammalian target-of-rapamycin growth and survival signaling in human hepatoma cells. Am. J. Physiol. Cell Physiol. 293, C55–C63 (2007).
Sundrud, M. S. et al. Halofuginone inhibits TH17 cell differentiation by activating the amino acid starvation response. Science 324, 1334–1338 (2009).
Nicklin, P. et al. Bidirectional transport of amino acids regulates mTOR and autophagy. Cell 136, 521–534 (2009).
Findlay, J. S. & Ulaeto, D. Semliki Forest virus and Sindbis virus, but not vaccinia virus, require glycolysis for optimal replication. J. Gen. Virol. 96, 2693–2696 (2015).
Fontaine, K. A., Sanchez, E. L., Camarda, R. & Lagunoff, M. Dengue virus induces and requires glycolysis for optimal replication. J. Virol. 89, 2358–2366 (2015).
Fontaine, K. A., Camarda, R. & Lagunoff, M. Vaccinia virus requires glutamine but not glucose for efficient replication. J. Virol. 88, 4366–4374 (2014).
Li, C. Y., Wang, Y. J., Huang, S. W., Cheng, C. S. & Wang, H. C. Replication of the shrimp virus wssv depends on glutamate-driven anaplerosis. PLoS ONE 11, e0146902 (2016).
Thai, M. et al. MYC-induced reprogramming of glutamine catabolism supports optimal virus replication. Nat. Commun. 6, 8873 (2015).
Yu, Y., Clippinger, A. J. & Alwine, J. C. Viral effects on metabolism: changes in glucose and glutamine utilization during human cytomegalovirus infection. Trends Microbiol. 19, 360–367 (2011).
Chambers, J. W., Maguire, T. G. & Alwine, J. C. Glutamine metabolism is essential for human cytomegalovirus infection. J. Virol. 84, 1867–1873 (2010).
Loisel-Meyer, S. et al. Glut1-mediated glucose transport regulates HIV infection. Proc. Natl Acad. Sci. USA 109, 2549–2554 (2012).
Palmer, C. S. et al. Increased glucose metabolic activity is associated with CD4+ T-cell activation and depletion during chronic HIV infection. AIDS 28, 297–309 (2014).
Kavanagh Williamson, M. et al. Upregulation of glucose uptake and hexokinase activity of primary human CD4+ T cells in response to infection with HIV-1. Viruses 10, 114 (2018).
Hegedus, A., Kavanagh Williamson, M. & Huthoff, H. HIV-1 pathogenicity and virion production are dependent on the metabolic phenotype of activated CD4+ T cells. Retrovirology 11, 98 (2014).
Palmer, C. S. et al. Metabolically active CD4+ T cells expressing Glut1 and OX40 preferentially harbor HIV during in vitro infection. FEBS Lett. 591, 3319–3332 (2017).
Hegedus, A. et al. Evidence for altered glutamine metabolism in human immunodeficiency virus type 1 infected primary human CD4+ T cells. AIDS Res. Hum. Retroviruses 33, 1236–1247 (2017).
Macintyre, A. N. et al. The glucose transporter Glut1 is selectively essential for CD4 T cell activation and effector function. Cell Metab. 20, 61–72 (2014).
Xiao, Nakaya,M. et al. Inflammatory T cell responses rely on amino acid transporter ASCT2 facilitation of glutamine uptake and mTORC1 kinase activation. Immunity 40, 692–705 (2014).
Sinclair, L. V. et al. Control of amino-acid transport by antigen receptors coordinates the metabolic reprogramming essential for T cell differentiation. Nat. Immunol. 14, 500–508 (2013).
Hukelmann, J. L. et al. The cytotoxic T cell proteome and its shaping by the kinase mTOR. Nat. Immunol. 17, 104–112 (2016).
Cretenet, G. et al. Cell surface Glut1 levels distinguish human CD4 and CD8 T lymphocyte subsets with distinct effector functions. Sci. Rep. 6, 24129 (2016).
Buck, M. D., Sowell, R. T., Kaech, S. M. & Pearce, E. L. Metabolic instruction of immunity. Cell 169, 570–586 (2017).
Loftus, R. M. & Finlay, D. K. Immunometabolism: cellular metabolism turns immune regulator. J. Biol. Chem. 291, 1–10 (2016).
Chang, C. H. et al. Posttranscriptional control of T cell effector function by aerobic glycolysis. Cell 153, 1239–1251 (2013).
Lane, A. N. & Fan, T. W. Regulation of mammalian nucleotide metabolism and biosynthesis. Nucleic Acids Res. 43, 2466–2485 (2015).
Korin, Y. D. & Zack, J. A. Nonproductive human immunodeficiency virus type 1 infection in nucleoside-treated G0 lymphocytes. J. Virol. 73, 6526–6532 (1999).
Baldauf, H. M. et al. SAMHD1 restricts HIV-1 infection in resting CD4+ T cells. Nat. Med. 18, 1682–1687 (2012).
Lahouassa, H. et al. SAMHD1 restricts the replication of human immunodeficiency virus type 1 by depleting the intracellular pool of deoxynucleoside triphosphates. Nat. Immunol. 13, 223–228 (2012).
Craveiro, M., Clerc, I., Sitbon, M. & Taylor, N. Metabolic pathways as regulators of HIV infection. Curr. Opin. HIV AIDS 8, 182–189 (2013).
Carr, E. L. et al. Glutamine uptake and metabolism are coordinately regulated by ERK/MAPK during T lymphocyte activation. J. Immunol. 185, 1037–1044 (2010).
Wang, R. et al. The transcription factor Myc controls metabolic reprogramming upon T lymphocyte activation. Immunity 35, 871–882 (2011).
Klysz, D. et al. Glutamine-dependent alpha-ketoglutarate production regulates the balance between T helper 1 cell and regulatory T cell generation. Sci. Signal. 8, ra97 (2015).
Laplante, M. & Sabatini, D. M. An emerging role of mTOR in lipid biosynthesis. Curr. Biol. 19, R1046–R1052 (2009).
Laplante, M. & Sabatini, D. M. mTOR signaling at a glance. J. Cell. Sci. 122, 3589–3594 (2009).
Sengupta, S., Peterson, T. R. & Sabatini, D. M. Regulation of the mTOR complex 1 pathway by nutrients, growth factors, and stress. Mol. Cell 40, 310–322 (2010).
Nakaya, M. et al. Inflammatory T cell responses rely on amino acid transporter ASCT2 facilitation of glutamine uptake and mTORC1 kinase activation. Immunity 40, 692–705 (2014).
Billiard, J. et al. Quinoline 3-sulfonamides inhibit lactate dehydrogenase A and reverse aerobic glycolysis in cancer cells. Cancer Metab. 1, 19 (2013).
Duran, R. V. et al. Glutaminolysis activates Rag-mTORC1 signaling. Mol. Cell 47, 349–358 (2012).
Cavrois, M., De Noronha, C. & Greene, W. C. A sensitive and specific enzyme-based assay detecting HIV-1 virion fusion in primary T lymphocytes. Nat. Biotechnol. 20, 1151–1154 (2002).
Mamede, J. I. & Hope, T. J. Detection and tracking of dual-labeled HIV particles using wide-field live cell imaging to follow viral core integrity. Methods Mol. Biol. 1354, 49–59 (2016).
Fendt, S. M. et al. Reductive glutamine metabolism is a function of the alpha-ketoglutarate to citrate ratio in cells. Nat. Commun. 4, 2236 (2013).
Mullen, A. R. et al. Oxidation of alpha-ketoglutarate is required for reductive carboxylation in cancer cells with mitochondrial defects. Cell Rep. 7, 1679–1690 (2014).
Almeida, L., Lochner, M., Berod, L. & Sparwasser, T. Metabolic pathways in T cell activation and lineage differentiation. Semin. Immunol. 28, 514–524 (2016).
Yong, C. S. et al. Metabolic orchestration of T lineage differentiation and function. FEBS Lett. 591, 3104–3118 (2017).
Valle-Casuso, J. C. et al. Cellular metabolism is a major determinant of HIV-1 reservoir seeding in CD4+ T cells and offers an opportunity to tackle infection. Cell Metab. 29, 611–626 e615 (2019).
Devadas, S., Zaritskaya, L., Rhee, S. G., Oberley, L. & Williams, M. S. Discrete generation of superoxide and hydrogen peroxide by T cell receptor stimulation: selective regulation of mitogen-activated protein kinase activation and fas ligand expression. J. Exp. Med. 195, 59–70 (2002).
Kwon, J. et al. The nonphagocytic NADPH oxidase Duox1 mediates a positive feedback loop during T cell receptor signaling. Sci. Signal. 3, ra59 (2010).
Ron-Harel, N. et al. Mitochondrial biogenesis and proteome remodeling promote one-carbon metabolism for T cell activation. Cell Metab. 24, 104–117 (2016).
Sena, L. A. et al. Mitochondria are required for antigen-specific T cell activation through reactive oxygen species signaling. Immunity 38, 225–236 (2013).
van der Windt, G. J. et al. CD8 memory T cells have a bioenergetic advantage that underlies their rapid recall ability. Proc. Natl Acad. Sci. USA 110, 14336–14341 (2013).
Karniely, S. et al. Human cytomegalovirus infection upregulates the mitochondrial transcription and translation machineries. MBio 7, e00029 (2016).
Bukrinskaya, A., Brichacek, B., Mann, A. & Stevenson, M. Establishment of a functional human immunodeficiency virus type 1 (HIV-1) reverse transcription complex involves the cytoskeleton. J. Exp. Med. 188, 2113–2125 (1998).
Mandal, D. & Prasad, V. R. Analysis of 2-LTR circle junctions of viral DNA in infected cells. Methods Mol. Biol. 485, 73–85 (2009).
Laguette, N. et al. SAMHD1 is the dendritic- and myeloid-cell-specific HIV-1 restriction factor counteracted by Vpx. Nature 474, 654–657 (2011).
Hrecka, K. et al. Vpx relieves inhibition of HIV-1 infection of macrophages mediated by the SAMHD1 protein. Nature 474, 658–661 (2011).
Descours, B. et al. SAMHD1 restricts HIV-1 reverse transcription in quiescent CD4+ T-cells. Retrovirology 9, 87 (2012).
Matheson, N. J. et al. Cell surface proteomic map of HIV infection reveals antagonism of amino acid metabolism by Vpu and Nef. Cell Host Microbe 18, 409–423 (2015).
Matheson, N. et al. Antagonism of aminoacid transport in primary CD4 T cells by HIV-1 Vpu. Lancet 385, S66 (2015).
Michalek, R. D. et al. Cutting edge: distinct glycolytic and lipid oxidative metabolic programs are essential for effector and regulatory CD4+ T cell subsets. J. Immunol. 186, 3299–3303 (2011).
Shi, L. Z. et al. HIF1α-dependent glycolytic pathway orchestrates a metabolic checkpoint for the differentiation of TH17 and Treg cells. J. Exp. Med. 208, 1367–1376 (2011).
Berod, L. et al. De novo fatty acid synthesis controls the fate between regulatory T and T helper 17 cells. Nat. Med. 20, 1327–1333 (2014).
Gerriets, V. A. et al. Metabolic programming and PDHK1 control CD4+ T cell subsets and inflammation. J. Clin. Invest. 125, 194–207 (2015).
Franchi, L. et al. Inhibiting oxidative phosphorylation in vivo restrains Th17 effector responses and ameliorates murine colitis. J. Immunol. 198, 2735–2746 (2017).
Downs-Canner, S. et al. Suppressive IL-17A+Foxp3+ and ex-Th17 IL-17AnegFoxp3+ Treg cells are a source of tumour-associated Treg cells. Nat. Commun. 8, 14649 (2017).
Monteiro, P. et al. Memory CCR6+CD4+ T cells are preferential targets for productive HIV type 1 infection regardless of their expression of integrin β7. J. Immunol. 186, 4618–4630 (2011).
Gosselin, A. et al. Peripheral blood CCR4+CCR6+ and CXCR3+CCR6+CD4+ T cells are highly permissive to HIV-1 infection. J. Immunol. 184, 1604–1616 (2010).
Manel, N. et al. The ubiquitous glucose transporter GLUT-1 Is a receptor for HTLV. Cell 115, 449–459 (2003).
Kim, F. J. et al. HTLV-1 and -2 envelope SU subdomains and critical determinants in receptor binding. Retrovirology 1, 41 (2004).
Swainson, L. et al. Glucose transporter 1 expression identifies a population of cycling CD4+ CD8+ human thymocytes with high CXCR4-induced chemotaxis. Proc. Natl Acad. Sci. USA 102, 12867–12872 (2005).
Kinet, S. et al. Isolated receptor binding domains of HTLV-1 and HTLV-2 envelopes bind Glut-1 on activated CD4+ and CD8+ T cells. Retrovirology 4, 31 (2007).
Verhoeyen, E. et al. IL-7 surface-engineered lentiviral vectors promote survival and efficient gene transfer in resting primary T lymphocytes. Blood 101, 2167–2174 (2003).
Oburoglu, L. et al. Glucose and glutamine metabolism regulate human hematopoietic stem cell lineage specification. Cell Stem Cell 15, 169–184 (2014).
We thank all members of our laboratories for discussions and scientific critique and are indebted to S. Kinet and V. Zimmermann for their continual support in this project. We are indebted to E. Gottlieb for his important input into metabolic experiments and to J. Mamede for his expertise and input into viral fusion assays. C. Goujon and C. June generously provided reagents, as indicated. We are grateful to M. Boyer and S. Gailhac of Montpellier Rio Imaging for support in cytometry experiments. I.C. and D.A.M. were supported by fellowships from Sidaction. Z.V. was supported by a fellowship from the Fondation de la Recherche Medicale (FRM). S.T. is supported by funding from Cancer Research UK (C596/A17196, Award 23982). L.O. was supported by a fellowship from the Ligue Contre le Cancer and the ARC Foundation. M.S. and N.T. are supported by Inserm and V.D. and C.M by the CNRS. This work was supported by generous funding from Sidaction, the ANRS, ARC, FRM, the French national (ANR) research grants (PolarATTACK and GlutStem) and the French laboratory consortiums (Labex) EpiGenMed and GR-EX.
M.S. and N.T. are inventors on patent WO2010079208. M.S., C.M. and N.T. are inventors on patent WO/2004/096841 and M.S. is an inventor on patent WO/2012/035369. All patents are owned by the CNRS and cover the use of RBD ligands for metabolite transporter detection. N.T. no longer owns any patent rights. M.S. is a co-founder of METAFORA Biosystems.
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Clerc, I., Abba Moussa, D., Vahlas, Z. et al. Entry of glucose- and glutamine-derived carbons into the citric acid cycle supports early steps of HIV-1 infection in CD4 T cells. Nat Metab 1, 717–730 (2019). https://doi.org/10.1038/s42255-019-0084-1
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