IL-12 Produced by Dendritic Cells Augments CD8 + T Cell Activation through the Production of the Chemokines CCL1 and CCL17 This information is current as of September 23, 2021. Curtis J. Henry, David A. Ornelles, Latoya M. Mitchell, Kristina L. Brzoza-Lewis and Elizabeth M. Hiltbold J Immunol 2008; 181:8576-8584; ; doi: 10.4049/jimmunol.181.12.8576 http://www.jimmunol.org/content/181/12/8576 Downloaded from References This article cites 75 articles, 35 of which you can access for free at: http://www.jimmunol.org/content/181/12/8576.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 • Fast Publication! 4 weeks from acceptance to publication by guest on September 23, 2021 *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 © 2008 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology IL-12 Produced by Dendritic Cells Augments CD8؉ T Cell Activation through the Production of the Chemokines CCL1 and CCL171 Curtis J. Henry, David A. Ornelles, Latoya M. Mitchell, Kristina L. Brzoza-Lewis, and Elizabeth M. Hiltbold2 IL-12 family members are an important link between innate and adaptive immunity. IL-12 drives Th1 responses by augmenting IFN-␥ production, which is key for clearance of intracellular pathogens. IL-23 promotes the development of IL-17-producing CD4؉ T cells that participate in the control of extracellular pathogens and the induction of autoimmunity. However, recent studies have shown that these cytokines can modulate lymphocyte migration and cellular interactions. Therefore, we sought to determine ؉ the individual roles of IL-12 and IL-23 in naive CD8 T cell activation by addressing their ability to influence IFN-␥ production Downloaded from and cellular interaction dynamics during priming by Listeria monocytogenes-infected dendritic cells (DC). We found that IL-12 .was the major cytokine influencing the level of IFN-␥ production by CD8؉ T cells while IL-23 had little effect on this response In addition, we observed that IL-12 promoted longer duration conjugation events between CD8؉ T cells and DC. This enhanced cognate interaction time correlated with increased production of the chemokines CCL1 and CCL17 by WT but not IL-12-deficient DC. Neutralization of both chemokines resulted in reduced interaction time and IFN-␥ production, demonstrating their impor- ؉ ؉ tance in priming naive CD8 T cells. Our study demonstrates a novel mechanism through which IL-12 augments naive CD8 T http://www.jimmunol.org/ cell activation by facilitating chemokine production, thus promoting more stable cognate interactions during priming. The Journal of Immunology, 2008, 181: 8576–8584. t is becoming increasingly clear that the initial signals CD8ϩ DC that promote long-term physical interactions leading to the T cells receive during priming impacts the subsequent pri- efficient priming of naive CD8ϩ T cells remains to be determined. mary and secondary responses generated in these cells (1– IL-12 is produced by mature DC in response to infection by I 3 13). Dendritic cells (DC) efficiently deliver the required signals to various intracellular pathogens (14, 22). Due to the potent effects ϩ activate naive CD8 T cells. Without DC, adaptive immune re- that IL-12 has on T cell activation, it is largely viewed as an im- by guest on September 23, 2021 sponses against several pathogens are not elicited (14–17). Their portant bridge between the innate and adaptive immune systems potent ability to stimulate naive T cells depends on their matura- (23–29). This cytokine influences several aspects of T cell activa- tion state (14, 17). Mature DC will present several activating li- tion, most notably IFN-␥ production, which plays a key role in the ϩ gands or secreted factors to naive CD8 T cells during priming resolution of intracellular pathogens such as Listeria monocyto- such as peptide-MHC complexes (signal 1), costimulatory mole- ϩ genes (Lm) (30, 31). Extensive research has been performed on cules (signal 2), and cytokines (signal 3) (18–20). CD8 T cells Lm and the immune responses required for resolving infection are encountering mature DC engage in long duration interactions, re- well documented (32). IL-12 is required to resolve high-dose in- sulting in the generation of highly functional effector cells (21). fection; however, in the absence of IL-12, increased IFN-␥ can However, an immature DC has a reduced capacity to prime naive ϩ compensate (30, 33, 34). Direct infection of DC with Lm in vitro CD8 T cells which correlates with shorter duration interactions results in production of p40, a cytokine subunit shared by IL-12 during activation (21). The full complement of factors delivered by and IL-23 (14). Furthermore, neutralization of p40 during priming of CD8ϩ T cells by Listeria-infected DC results in a significant decrease in the amount of IFN-␥ produced by these cells (14). Department of Microbiology and Immunology, Wake Forest University School of IFN-␥ plays multiple roles in resolving listerial infection including Medicine, Winston-Salem, NC 27157 activating macrophages and increasing target cell sensitivity to ly- Received for publication March 5, 2008. Accepted for publication October 13, 2008. sis by CD8ϩ T cells (8, 30, 35, 36). Thus, IL-12 production is a The costs of publication of this article were defrayed in part by the payment of page key hallmark of resolving Lm infection through its augmentation charges. This article must therefore be hereby marked advertisement in accordance of IFN-␥ production. with 18 U.S.C. Section 1734 solely to indicate this fact. The IL-12 family is composed of the cytokines IL-12, IL-23, and 1 This research was supported by the National Institutes of Health Research Project Grant Program (R01) Grant AI057770-01A1 (to E.M.H.). Additional support was IL-27 (reviewed in Refs. 24–29, 37–39). They are grouped together provided by the National Institutes of Health Predoctoral Fellowship Award for Mi- based on structural similarities between the cytokines and/or their nority Students (F31) Grant AI73245-01A1 (to C.J.H.). receptors. IL-12 and IL-23 share the p40 subunit. However, p35 is 2 Address correspondence and reprint requests to Dr. Elizabeth M. Hiltbold, Depart- ment of Microbiology and Immunology, 5109 Gray Building, Wake Forest University exclusively associated with IL-12 and a heterodimer of p19/p40 School of Medicine, Winston-Salem, NC 27157. E-mail address: bhiltbol@ makes up IL-23. The distinct biological functions of each cytokine wfubmc.edu in this family may be attributed to differences in the composition 3 Abbreviations used in this paper: DC, dendritic cell; Lm, Listeria monocytogenes; of their receptors and distinct sets of downstream transcription WT, wild type; MFI, mean fluorescence intensity; ICS, intracellular cytokine staining. factors that are activated upon ligation. The main role attributed to Copyright © 2008 by The American Association of Immunologists, Inc. 0022-1767/08/$2.00 IL-12 is the ability to induce and augment IFN-␥ production by www.jimmunol.org The Journal of Immunology 8577 ϩ CD4 T cells (driving a Th1-type response) as well as NK and i.v. (5 ϫ 105). At 18 h after infection, mice were sacrificed and inguinal, CD8ϩ T cells. IL-12 has been shown to increase CD8ϩ T cell lumbar, and mesenteric lymph nodes as well as spleens were harvested. RNA was isolated for chemokine analysis from DC following enrichment cytolysis, survival, proliferation, and influence the ability of these ϩ for CD11c cells. lymphocytes to migrate to inflammatory foci. A more recently rec- ognized activity of IL-23 is the ability to stimulate the production In vitro infection of DC ϩ of IL-17 by CD4 T cells. These Th17 cells are critical for the For T cell priming assays, DC were seeded at 2 ϫ 104/well and infected development of autoimmune diseases and immunity to certain ex- with WT Listeria (strain 10403S) at a multiplicity of infection (MOI) of 1. tracellular pathogens (24–29, 37–45). However, the role of IL-23 Four hours after infection, OVA peptide257–264 was added at a concentra- ␮ in the activation of naive CD8ϩ T cells is still being investigated. tion of 0.1 ng/ml (or as indicated) with chloramphenicol (10 g/ml) and gentamicin (10 ␮g/ml). Twenty-four hours after infection, OT-I T cells Since IL-12 and IL-23 have very potent effects on specific were added at 10:1 T:DC. For some experiments, OT-I T cells were stained immune responses, we wanted to determine the individual con- with CFSE before culture with DC. ϩ tributions of IL-12 and IL-23 on CD8 T cell priming by DC. Time-lapse video microscopy In addition, we wanted to identify the mechanism(s) through which these cytokines modulated naive CD8ϩ T cell activation. DC were prepared and infected as in T cell priming assays with the fol- lowing modifications. Day 6 WT, IL-12p35Ϫ/Ϫ, or IL-12p40Ϫ/Ϫ DC (106) To approach these goals, we used DC generated from mice Ϫ/Ϫ Ϫ/Ϫ were seeded in T25 flask and infected with Lm at a MOI of 1. After 4 h, lacking both IL-12 and IL-23 (p40 ) or only IL-12 (p35 ).