Dendritic Cells Support Sequential Reprogramming of Chemoattractant Receptor Profiles During Naive to Effector T Cell Differentiation This information is current as of September 25, 2021. Chang H. Kim, Kinya Nagata and Eugene C. Butcher J Immunol 2003; 171:152-158; ; doi: 10.4049/jimmunol.171.1.152 http://www.jimmunol.org/content/171/1/152 Downloaded from References This article cites 52 articles, 27 of which you can access for free at: http://www.jimmunol.org/content/171/1/152.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 25, 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 © 2003 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Dendritic Cells Support Sequential Reprogramming of Chemoattractant Receptor Profiles During Naive to Effector T Cell Differentiation1 Chang H. Kim,2* Kinya Nagata,† and Eugene C. Butcher‡ T cells undergo chemokine receptor switches during activation and differentiation in secondary lymphoid tissues. Here we present evidence that dendritic cells can induce changes in T cell expression of chemokine receptors in two continuous steps. In the first switch over a 4–5 day period, dendritic cells up-regulate T cell expression of CXCR3 and CXCR5. Additional stimulation leads to the second switch: down-regulation of lymphoid tissue homing related CCR7 and CXCR5, and up-regulation of Th1/2 effector tissue-targeting chemoattractant receptors such as CCR4, CCR5, CXCR6, and CRTH2. We show that IL-4 and IL-12 can determine the fate of the secondary chemokine receptor switch. IL-4 enhances the generation of CCR4؉ and CRTH2؉ T cells, and Downloaded from .suppresses the generation of CXCR3؉ T cells and CCR7؊ T cells, while IL-12 suppresses the level of CCR4 in responding T cells Furthermore, IL-4 has positive effects on generation of CXCR5؉ and CCR7؉ T cells during the second switch. Our study suggests that the sequential switches in chemokine receptor expression occur during naive T cell interaction with dendritic cells. The first switch of T cell chemokine receptor expression is consistent with the fact that activated T cells migrate within lymphoid tissues for interaction with B and dendritic cells, while the second switch predicts the trafficking behavior of effector T cells away from lymphoid tissues to effector tissue sites. The Journal of Immunology, 2003, 171: 152–158. http://www.jimmunol.org/ eterogeneity in lymphocyte trafficking behavior is deter- moattractant receptors induced or down-regulated? This would be mined by expression patterns of chemokine receptors valuable information in understanding changes in the trafficking H and adhesion molecules (1–4). Chemokines regulate behavior of the T cells undergoing activation and differentiation at leukocyte trafficking by inducing firm integrin-dependent adhesion the early vs late stages of immune responses. of blood leukocytes to endothelial cells, and by inducing direc- Dendritic cells capture Ags, mature, and up-regulate CCR7 in tional migration (chemotaxis). Leukocytes express over 20 che- inflamed tissue sites (23, 24). Mature dendritic cells migrate into mokine or chemoattractant receptors in both subset-specific and the T cell areas of secondary lymphoid tissues via afferent lym- overlapping patterns. Recently, there has been significant progress phatic vessels. Circulating naive T cells are also programmed to by guest on September 25, 2021 in characterization of chemokine receptors expressed by T cells at migrate into the T cell area of the secondary lymphoid tissues various developmental stages and functional status: immature and through a specialized endothelial layer called high endothelial mature thymocytes express different sets of chemokine receptors venules. Endothelial and other cell types in the T cell areas express (5–9); naive and memory effector T cells are also different in ex- specific adhesion and chemokine molecules (e.g., peripheral node pression of chemokine receptors (10–20). Th1 and Th2 cells are addressin and secondary lymphoid tissue chemokine (CCL21)/ distinguished from each other in expression of several chemokine EBI1-ligand chemokine (CCL19) in peripheral lymph nodes) and receptors (1, 10, 12, 13, 15, 18, 21, 22). Despite overlapping ex- recruit naive T cells into lymphoid tissues (25, 26). Dendritic cells pression patterns, chemokine receptors are often classified for sim- in the T cell area present Ag peptides to naive T cells. T cells then plicity as Th1 (CXCR3, CCR5, and CXCR6) vs Th2 (CCR4, undergo activation processes and differentiate into memory and CCR3, and CCR8) or lymphoid homing (CCR7 and CXCR5) vs effector T cell subsets with specialized functions (e.g., Th1 and non-lymphoid tissue homing (Th1 and Th2 types combined) types. Th2 cells (23, 24) or specialized gut vs skin-targeted cells (27, An important unanswered question is at which time during the 28)). differentiation of T cells is the expression of chemokine or che- Although dendritic cells have been well demonstrated to be crit- ical in activating and differentiating T cells, it remains to be sys- tematically determined when and how different types of chemo- *Laboratory of Immunology and Hematopoiesis, Department of Veterinary Pathobi- kine receptors are regulated on T cells undergoing differentiation ology and Purdue Cancer Center, and Biochemistry and Molecular Biology Program, to memory and effector T cells in response to signals of dendritic Purdue University, West Lafayette, IN 47907; †R & D Center, BML, Saitama, Japan; ‡Laboratory of Immunology and Vascular Biology, Department of Pathology, Stan- cells. We found that dendritic cells and their cytokines play crucial ford University School of Medicine, Stanford, CA 94305, and Center for Molecular roles in sequential switching of chemokine receptors from naive to Biology and Medicine, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA 94304 early memory, and then to effector types. These changes are im- portant for the stage-specific migration and/or interaction of T cells Received for publication December 5, 2002. Accepted for publication April 18, 2003. with other cell types. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. Materials and Methods 1 This work was supported by grants from The Eli and Edythe L. Broad Foundation, Abs and cytokines the Leukemia and Lymphoma Society (to C.H.K.), and the National Institute of Health (to E.C.B.). Abs to CD4 (RPA-T4), CD11C (B-ly6), CD45RA (HI100), CD45RO 2 Address correspondence and reprint requests to Dr. Chang H. Kim, 1243 VPTH, Purdue (UCHL1) and IFN-␥ (4S.B3) were purchased from BD PharMingen (San University, West Lafayette, IN 47907-1243. E-mail address: [email protected] Diego, CA). Anti-IL4-PE (3010.211) was purchased from BD Biosciences Copyright © 2003 by The American Association of Immunologists, Inc. 0022-1767/03/$02.00 The Journal of Immunology 153 (San Jose, CA). Abs to CXCR6 (56811.111), CCR1 (53504.111), CCR2 CD45RA, CD4, and/or chemokine receptors) for cell surface Ags, then (48607.121), CCR5 (45549.111), CCR6 (53103.111), CXCR3 activated for4hat37°C with PMA (50 ng/ml) and ionomycin (1 ␮g/ml) (49801.111), and CXCR5 (51505.111) were obtained from R&D Systems in RPMI 1640 medium supplemented with penicillin/streptomycin, 10% (Minneapolis, MN). Abs to CCR3 (7B11) and CCR9 (GPR96-1) were FBS, and 10 ␮g/ml Monensin (Sigma-Aldrich). Activated cells were fixed purchased from Millennium Pharmaceuticals (Cambridge, MA). Abs to and permeabilized using Cytofix/Cytoperm solution (PharMingen) and CCR4 (1G1) and CCR7 (7H12), and recombinant human IL-4 and IL-12 stained with isotype control Abs or mAbs to IL-4 (PE) and IFN-␥ (FITC). were provided by BD PharMingen. Four color flow cytometry was done on a FACSCalibur (BD Biosciences) using CellQuest software, version 3.1 (BD Biosciences), or Cytomics Cell isolation and preparation FC500 (Beckman Coulter, Fullerton, CA), and WinMDI (v2.8) software. PBMC from human peripheral blood (Indiana Regional Blood Center, In- Statistical analyses dianapolis, IN) was prepared by density gradient centrifuge on Histopaque- ϩ 1077 (Sigma-Aldrich, St. Louis, MO). CD4 T cells (purity Ͼ97%) were Student’s t test was used. Values of p Ͼ 0.05 were considered to be sig- isolated by depleting non-T cells using a magnetic bead depletion method nificant differences. (Miltenyi Biotec, Auburn, CA). Naive CD45RAϩ CD4 T cells were further sorted by FACSVantage SE (BD Biosciences, purity Ͼ99.5%) or by CD45ROϩ cell depletion by MACS. All human subject protocols were Results approved by the Institutional Review Board at Purdue University (West Dendritic cells induce chemokine receptor switch Lafayette, IN). Peripheral blood CD14ϩ monocytes (purity Ͼ99%) were isolated by magnetic sorting (Miltenyi Biotec). Immature dendritic cells We generated mature dendritic cells from peripheral monocytes, were generated by culturing CD14ϩ monocytes for 5 days in RPMI 1640 and examined whether dendritic cells can up-regulate chemokine medium (10% FBS), supplemented with IL-4 (1000 U/ml, BD PharMin- receptors on T cells undergoing differentiation. T cells were ex- gen) and GM-CSF (50 ng/ml, R&D Systems). After the initial culture, all Downloaded from Ϫ ϩ amined for expression of chemokine receptors after 4 days in co- immature dendritic cells show a phenotype of CD14 CD11C .