Early Inhibition of Fatty Acid Synthesis Reduces Generation of Memory Precursor Effector T Cells in Chronic Infection This information is current as Samad A. Ibitokou, Brian E. Dillon, Mala Sinha, Bartosz of October 1, 2021. Szczesny, Añahi Delgadillo, Doaa Reda Abdelrahman, Csaba Szabo, Lutfi Abu-Elheiga, Craig Porter, Demidmaa Tuvdendorj and Robin Stephens J Immunol 2018; 200:643-656; Prepublished online 13 December 2017; doi: 10.4049/jimmunol.1602110 Downloaded from http://www.jimmunol.org/content/200/2/643 Supplementary http://www.jimmunol.org/content/suppl/2017/12/13/jimmunol.160211 http://www.jimmunol.org/ Material 0.DCSupplemental References This article cites 69 articles, 21 of which you can access for free at: http://www.jimmunol.org/content/200/2/643.full#ref-list-1 Why The JI? Submit online. by guest on October 1, 2021 • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2018 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Early Inhibition of Fatty Acid Synthesis Reduces Generation of Memory Precursor Effector T Cells in Chronic Infection Samad A. Ibitokou,* Brian E. Dillon,* Mala Sinha,† Bartosz Szczesny,‡ An˜ahi Delgadillo,x Doaa Reda Abdelrahman,x Csaba Szabo,‡ Lutfi Abu-Elheiga,{ Craig Porter,x Demidmaa Tuvdendorj,‖ and Robin Stephens*,# Understanding the mechanisms of CD4 memory T cell (Tmem) differentiation in malaria is critical for vaccine development. However, the metabolic regulation of CD4 Tmem differentiation is not clear, particularly in persistent infections. In this study, we investigated the role of fatty acid synthesis (FAS) in Tmem development in Plasmodium chabaudi chronic mouse malaria infection. We show that T cell–specific deletion and early pharmaceutical inhibition of acetyl CoA carboxylase 1, the rate limiting step of FAS, inhibit generation of early memory precursor effector T cells (MPEC). To compare the role of FAS during early differen- tiation or survival of Tmem in chronic infection, a specific inhibitor of acetyl CoA carboxylase 1, 5-(tetradecyloxy)-2-furoic acid, Downloaded from was administered at different times postinfection. Strikingly, the number of Tmem was only reduced when FAS was inhibited during T cell priming and not during the Tmem survival phase. FAS inhibition during priming increased effector T cell (Teff) proliferation and strongly decreased peak parasitemia, which is consistent with improved Teff function. Conversely, MPEC were decreased, in a T cell–intrinsic manner, upon early FAS inhibition in chronic, but not acute, infection. Early cure of infection also increased mitochondrial volume in Tmem compared with Teff, supporting previous reports in acute infection. We demonstrate that the MPEC-specific effect was due to the higher fatty acid content and synthesis in MPEC compared with terminally http://www.jimmunol.org/ differentiated Teff. In conclusion, FAS in CD4 T cells regulates the early divergence of Tmem from Teff in chronic infection. The Journal of Immunology, 2018, 200: 643–656. espite some progress in the control of malaria, the World chronic parasites (6). We have shown that CD4 T cells in the memory Health Organization estimates that 3.2 billion people are still phase of P. chabaudi infection primarily have an effector memory at high risk worldwide (1). The most recent subunit malaria T cell (Tem) phenotype and that these cells contain an increased D 2 vaccine candidate, RTS,S, has shown poor efficacy and a lack of proportion of IFN-g+TNF+IL-2 Th1 cells during chronic infection long-lived protection (2). Immunity to blood-stage malaria infection, compared with infections cured after 30 d (7). However, the mech- by guest on October 1, 2021 the stage of parasite that causes disease pathology, requires CD4 anisms of Tem differentiation are much less well understood than T cell– and B cell–mediated mechanisms (3). Yet, mechanisms of those of central memory T cells (Tcm) generated in acute stimulation. activation and differentiation for these protective cells are not well Two models have been proposed for the differentiation of understood. Infection of mice with Plasmodium chabaudi is an ac- memory T cells (Tmem). In simplified terms, these models propose curate and well-defined model for the immunity and pathology of a bifurcating model in which all Tmem, including Tem, are gen- mild malaria, and it has a chronic phase lasting up to 3 mo (4). erated early in activation (8) or a linear model in which Tem are Plasmodium falciparum is also documented to become chronic, derived from effector T cells (Teff) and are predicted to have a lasting up to a year, even in the absence of a rainy season and short half-life (9). It should be noted that early differentiation mosquitos (5). Chronic infection and chronically stimulated T cells steps do not exclude the role of later inflammatory effects that protect animals from reinfection against Plasmodium and other promote terminal differentiation in CD8 T cells or the role of *Division of Infectious Diseases, Department of Internal Medicine, University of Address correspondence and reprint requests to Dr. Robin Stephens, Division of Texas Medical Branch, Galveston, TX 77555; †Biomedical Informatics, Institute Infectious Diseases, Department of Internal Medicine and Department of Microbiol- for Translational Science, University of Texas Medical Branch, Galveston, TX ogy and Immunology, University of Texas Medical Branch, 301 University Boule- 77555; ‡Department of Anesthesiology, University of Texas Medical Branch, Gal- vard, Galveston, TX 77555-0435. E-mail address: [email protected] veston, TX 77555; xShriners Hospital for Children, Galveston, TX 77550; {Verna and The online version of this article contains supplemental material. Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030; ‖Division of Endocrinology, University of Texas Abbreviations used in this article: ACC, actyl CoA carboxylase; Cpt, carnitine pal- Medical Branch, Galveston, TX 77555; and #Department of Microbiology and Im- mitoyl transferase; CT, threshold cycle; CTV, CellTrace Violet; DAG, diacylglycer- munology, University of Texas Medical Branch, Galveston, TX 77555 ide; ECAR, extracellular respiratory capacity; ETC, electron transport chain; FAO, fatty acid oxidation; FAS, fatty acid synthesis; fDNL, fractional de novo lipogenesis; ORCIDs: 0000-0002-7760-7412 (S.A.I.); 0000-0003-3110-4235 (C.S.); 0000-0002- GC-MS, gas chromatograph–mass spectrometer; 2H O, deuterium water; MPEC, 7366-3556 (L.A.-E.); 0000-0002-5913-6194 (A.D.); 0000-0001-9847-5714 (D.R.A.). 2 memory precursor effector T cell; MQ, mefloquine hydrochloride; MSP-1, merozoite Received for publication December 15, 2016. Accepted for publication November 9, surface protein-1; MTG, MitoTracker Green; MTR, MitoTracker Red; NRx, no treat- 2017. ment; OCR, oxygen consumption rate; p.i., postinfection; PL, phospholipid; SLEC, short-lived effector T cell; SRC, spare respiratory capacity; Tcm, central memory This work was supported by National Institute of Allergy and Infectious Diseases T cell; Teff, effector T cell; TeffEarly, early Teff; TeffInt, intermediate Teff; TeffLate, Grant R01AI08995304 (to R.S.), the Department of Internal Medicine, University late Teff; Tem, effector memory T cell; TG, triglyceride; Tg, transgenic; Tmem, of Texas Medical Branch Institute for Human Infections and Immunity (to R.S. and memory T cell; TOFA, 5-(tetradecyloxy)-2-furoic acid; Treg, regulatory T cell; S.A.I.), and the American Association of Immunologists (to S.A.I.). WT, wild-type. The microarray data presented in this article have been submitted to the National Center for Biotechnology Information Gene Expression Omnibus (http://www.ncbi. Copyright Ó 2018 by The American Association of Immunologists, Inc. 0022-1767/18/$35.00 nlm.nih.gov/geo/query/acc.cgi?acc=GSE89555) under accession number GSE89555. www.jimmunol.org/cgi/doi/10.4049/jimmunol.1602110 644 EARLY CD4 Tmem DIFFERENTIATION DRIVEN BY FAS regulatory T cells (Treg) in controlling the quiescence of Tmem (9). generation of CD4 memory and support an early bifurcation in the Furthermore, there are differences between CD4 and CD8 Teff dif- differentiation of Tmem from Teff. ferentiation. For example, Blimp-1 drives terminal differentiation in CD8 Teff, but not in CD4 Teff (10, 11), which indicates that there are Materials and Methods likely differences in CD8 and CD4 Tmem differentiation as well. Our Mice and parasite work on CD4 T cell differentiation in P. chabaudi infection definitively B5 TCR–transgenic (Tg) mice, a kind gift from Jean Langhorne (The Francis support the early divergence of CD4 Teff and Tem in chronic infection Crick Institute, London, U.K.), were generated as previously described (23) and + 2 hi (12). We have recently identified CD4 IL-7Ra CD62L early Teff backcrossed to BALB/cJ mice (N4-N10; The Jackson Laboratory, Bar Harbor, (TeffEarly), which can be detected as early as day 5 postinfection (p.i.), ME). The B5 TCR recognizes merozoite surface protein-1 (MSP-1; 1157-1171, d as precursors of Tcm and Tem (12). We also showed that the Tem ISVLKSRLLKRKKYI/I-E ); B5 TCR–Tg mice were typed using primers Va2, 9 9 9 phenotype of the CD4+ Tmem developed in P. chabaudi infection is 5 -GAACGTTCCAGATTCCATGG-3 and 5 -ATGGACAAGATCCTGA- CAGCATCG-39 and Vb8.1, 59-CAGAGACCCTCAGGCGGCTGCTCAGG-39 and determined within the first 5 d of infection, because it can be blunted 59-ATGGGCTCCAGGCTGTTCTTTGTGGTTTTGATTC-39. Thy1.1 BALB/ by treatment of infection on day 3 p.i.