LAG-3 Confers a Competitive Disadvantage Upon Antiviral CD8+ T Cell Responses

LAG-3 Confers a Competitive Disadvantage upon Antiviral CD8 + T Cell Responses Kevin D. Cook and Jason K. Whitmire This information is current as J Immunol 2016; 197:119-127; Prepublished online 20 May of September 23, 2021. 2016; doi: 10.4049/jimmunol.1401594 http://www.jimmunol.org/content/197/1/119 Downloaded from Supplementary http://www.jimmunol.org/content/suppl/2016/05/20/jimmunol.140159 Material 4.DCSupplemental References This article cites 40 articles, 23 of which you can access for free at: http://www.jimmunol.org/content/197/1/119.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 23, 2021 • 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 © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology LAG-3 Confers a Competitive Disadvantage upon Antiviral CD8+ T Cell Responses Kevin D. Cook* and Jason K. Whitmire*,† Ongoing clinical trials are evaluating the benefits of systemic blockade of lymphocyte activation gene-3 (LAG-3) signals to improve immunity to tumors. Those studies are founded on the well-established inhibitory role of LAG-3 in regulating CD8+ T cells during chronic virus infection and antitumor responses. However, the T cell response in LAG-3–deficient mice is similar in size and function to that in wild type animals, suggesting LAG-3 has nuanced immune-regulatory functions. We performed a series of adoptive transfer experiments in mice to better understand the T cell–intrinsic functions of LAG-3 in the regulation of CD8+ T cell responses. Our results indicate that LAG-3 expression by CD8+ T cells inhibits their competitive fitness and results in a slightly reduced rate of cell division in comparison with LAG-3–deficient cells. This cell-intrinsic effect of LAG-3 was consistent across both acute and chronic virus infections. These data show that LAG-3 directly modulates the size of the T cell response and support Downloaded from the use of LAG-3 blockade regimens to enhance CD8+ T cell responses. The Journal of Immunology, 2016, 197: 119–127. hronic virus infections are a major health issue world- signaling (12, 13). During an acute infection, LAG-3 is expressed wide, with millions of people currently living with dis- early upon activation, perhaps because of its rapid translocation to C eases caused by viruses such as HIV, hepatitis C virus, the surface from a recycling endosomal compartment (14). Surface and hepatitis B virus. During these infections, T cell responses expression of LAG-3 can be efficiently reduced by two trans- http://www.jimmunol.org/ become functionally inactive through a process known as T cell membrane metalloproteases that cleave LAG-3 from the cell sur- + exhaustion and fail to clear the infection (1). Exhausted CD8 face (2, 15). Early studies highlighted the ability of LAG-3 to act as T cells show diminished cytokine production, weakened cytolytic an inhibitory receptor for CD8+ T cell expansion and homeostasis activity, and lower Ag-induced proliferation in large part because (16, 17). Further studies showed that surface LAG-3 expression is of their coexpression of a series of surface inhibitory receptors, maintained for an extended period during persistent virus infections, including programmed cell death protein 1 (PD-1), lymphocyte typically for as long as the Ag is present (2). The inhibitory function activation gene-3 (LAG-3; CD223), 2B4 (CD244), CD160, and of LAG-3 was confirmed by Ab blockade studies in which LAG-3 T cell Ig mucin-3 (Tim-3) (2). Recent data in mouse models in- interference led to enhanced T cell responses and improved virus dicate that T cell dysfunction can be reversed by combined Ab clearance (3). However, in some circumstances, the absence of by guest on September 23, 2021 blockade of these receptors, especially PD-1 and LAG-3, leading LAG-3 appears to have no effect on either T cell expansion or to improved clearance of viral and bacterial pathogens (3, 4). clearance of a persistent virus infection (18). Similar results have also been seen after blockade of LAG-3 in The discrepancies between these results could be partially tumor models, resulting in the elimination of tumors (5–7). There- because of the expression of LAG-3 by other cell types. Activated fore, it is important to understand the specific effects of LAG-3 on CD4+ T cells express LAG-3, which negatively regulates CD4+ CD8+ T cells to improve vaccines and the treatment of persistent T cell responses and homeostatic proliferation (16, 17, 19). infections and tumors. CD4+ regulatory T cells (Tregs) also express LAG-3 and lose LAG-3 is a surface receptor expressed by activated CD8+ T cells functionality in the absence of LAG-3 (20). LAG-3 deficiency leads that binds to MHC class II (MHC-II) with a higher affinity than to altered homeostatic regulation of plasmacytoid dendritic cells the T cell coreceptor CD4 (8–11). LAG-3 associates with the TCR at the immunological synapse after TCR stimulation and inhibits (pDCs), a key cell type in the production of type I IFN (21, 22). NK cells express LAG-3, and LAG-3–deficient mice have reduced NK cell activity (11, 23). There has also been a report doc- *Department of Genetics, University of North Carolina School of Medicine, Chapel Hill, NC 27599; and †Department of Microbiology and Immunology, University of umenting LAG-3 expression on B cells (24), although the func- North Carolina School of Medicine, Chapel Hill, NC 27599 tional significance of LAG-3 in B cell responses is not known. Received for publication July 1, 2014. Accepted for publication April 28, 2016. Thus, many cell types express LAG-3, and any or all of these cells + This work was supported by National Institutes of Health Grants R01-AI074862, could impact CD8 T cell responses. Because of this, the inter- R56-AI110682, and R21-AI117575 (to J.K.W.) and start-up funds from the Univer- pretation of data involving systemic Ab-mediated blockade of sity of North Carolina at Chapel Hill. LAG-3 must also take into account other LAG-3+ targets and their Address correspondence and reprint requests to Dr. Jason K. Whitmire, Department of Genetics, Department of Microbiology and Immunology, University of North regulatory effects. Carolina School of Medicine, 5062 Genetic Medicine Research Building, 120 Mason In this study, we addressed the cell-intrinsic role for LAG-3 Farm Road, Chapel Hill, NC 27599-7264. E-mail address: [email protected] expression on CD8+ T cell responses post acute and chronic virus The online version of this article contains supplemental material. infection. We found that LAG-3 expression by CD8+ T cells de- Abbreviations used in this article: cDC, conventional DC; DC, dendritic cell; ICS, creases the size of the virus-specific CD8+ Tcellresponsewithout intracellular cytokine staining; LAG-3, lymphocyte activation gene-3; LCMV, lymphocytic choriomeningitis virus; MHC-II, MHC class II; MPEC, memory-precursor inducing functional changes. We used competition experiments to effector cell; PD-1, programmed cell death protein 1; pDC, plasmacytoid dendritic reveal that LAG-3–deficient T cells outcompeted their LAG-3– cell; p.i., postinfection; Treg, regulatory T cell; WT, wild type. sufficient counterparts. Thus, LAG-3 has a cell-intrinsic negative + Copyright Ó 2016 by The American Association of Immunologists, Inc. 0022-1767/16/$30.00 effect on CD8 T cell responses that results in reduced fitness. www.jimmunol.org/cgi/doi/10.4049/jimmunol.1401594 120 LAG-3 INHIBITS CD8+ T CELLS Materials and Methods T cells in response to different variants of LCMV that result in acute Mice and virus (Armstrong) or persistent (Clone13) infection. LAG-3 was induced to high levels at day 5 with both variants. The percentage of LAG- C57BL/6 mice were purchased from Jackson Immunoresearch Laboratory. LAG-32/2 mice were originally generated by Drs. Diane Mathis and 3–expressing T cells returned to baseline by day 8 for Armstrong Christophe Benoist at the Institut Geńe´tique Biologie Molećulaire Cellu- (Fig. 1A); however, post Clone13 infection, LAG-3 expression was laire (Illkirch-Graffenstaden, France) and subsequently backcrossed to maintained for at least 60 d on a population of LCMV-specific CD8+ C57BL/6 by Dr. Dario Vignali at St. Jude Children’s Research Hospital T cells (Fig. 1A). The surface level of LAG-3 closely tracked with (Memphis, TN) and are a gift from Dr. Vignali and St. Jude Children’s Hospital (16, 23, 25). P14 TCR-transgenic mice that have T cells specific the amount of virus in the animals, consistent with previous studies for the lymphocytic choriomeningitis virus (LCMV) epitope GP33–41 were (2). LAG-3 is typically regarded as a negative regulator of T cell crossed to C57BL/6.Thy1.1 or C57BL/6.Ly5a mice to generate P14 Thy1.1+ responses, but multiple cell types can express LAG-3, including + + 2/2 and P14 Ly5a mice. The P14 Ly5a mice were crossed to LAG-3 mice CD4+ and CD8+ T cells, Tregs, B cells, pDCs, and NK cells (11, 20, to generate the P14 LAG-3–deficient TCR-transgenic mice.

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