Diverse and Potent Chemokine Production by Lung CD11b high Dendritic Cells in Homeostasis and in Allergic Lung Inflammation This information is current as of September 29, 2021. Steven R. Beaty, C. Edward Rose, Jr. and Sun-sang J. Sung J Immunol 2007; 178:1882-1895; ; doi: 10.4049/jimmunol.178.3.1882 http://www.jimmunol.org/content/178/3/1882 Downloaded from References This article cites 69 articles, 42 of which you can access for free at: http://www.jimmunol.org/content/178/3/1882.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 29, 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 © 2007 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Diverse and Potent Chemokine Production by Lung CD11bhigh Dendritic Cells in Homeostasis and in Allergic Lung Inflammation1 Steven R. Beaty, C. Edward Rose, Jr., and Sun-Sang J. Sung2 Lung CD11chigh dendritic cells (DC) are comprised of two major phenotypically distinct populations, the CD11bhigh DC and the ؉ ؉ ␤ ␣ integrin E 7 DC (CD103 DC). To examine whether they are functionally distinguishable, global microarray studies and real-time PCR analysis were performed. Significant differences between the two major CD11chigh DC types in chemokine mRNA expression were found. CD11bhigh DC is a major secretory cell type and highly expressed at least 16 chemokine mRNA in the homeostatic state, whereas CD103؉ DC highly expressed only 6. Intracellular chemokine staining of CD11chigh lung cells including macrophages, and ELISA determination of sort-purified CD11chigh cell culture supernatants, further showed that CD11bhigh DC Downloaded from produced the highest levels of 9 of 14 and 5 of 7 chemokines studied, respectively. Upon LPS stimulation in vitro and in vivo, CD11bhigh DC remained the highest producer of 7 of 10 of the most highly produced chemokines. Induction of airway hyperre- activity and lung inflammation increased lung CD11bhigh DC numbers markedly, and they produced comparable or higher amounts of 11 of 12 major chemokines when compared with macrophages. Although not a major producer, CD103؉ DC produced the highest amounts of the Th2-stimulating chemokines CCL17/thymus and activation-related chemokine and CCL22/monocyte- derived chemokine in both homeostasis and inflammation. Significantly, CCL22/monocyte-derived chemokine exhibited regula- http://www.jimmunol.org/ tory effects on CD4؉ T cell proliferation. Further functional analysis showed that both DC types induced comparable Th subset development. These studies showed that lung CD11bhigh DC is one of the most important leukocyte types in chemokine production .and it is readily distinguishable from CD103؉ DC in this secretory function. The Journal of Immunology, 2007, 178: 1882–1895 llergic asthma is caused by dysregulated immune re- expressed distinct surface markers from nonepithelial lung DC and sponses to airway allergens in house dust mites, molds, were more potent in Ag presentation than the latter DC type (12). and animal danders, which leads to a disease character- These early data suggest the occurrence of at least two major lung A 3 ized by airway hyperresponsiveness (AHR), reversible airway ob- DC types. In support of this occurrence, Flt3 ligand treatment was ϩ Ϫ struction, and lung inflammation (1, 2). Dendritic cells (DC) are found to increase a lung CD11b but not a CD11b DC popula- by guest on September 29, 2021 considered the major APC type in the capture, internalization, and tion preferentially, thus suggesting that CD11b may be considered transport of airway Ag to the lung-draining lymph node for Ag- a distinguishing marker (13). More detailed lung DC subset anal- specific T cell activation (3–7) and airway DC depletion abrogates ysis have confirmed the existence of CD11bhigh and the CD11blow Ag-induced AHR and lung inflammation (8, 9). Lung DC have populations (14). However, this marker was insufficient to differ- been shown to be occuring in or tightly apposed to the bronchial entiate the lung DC populations. CD103, an integrin protein sub- ␤ epithelium and DC extensions form a lattice-like pattern that line unit that associates only with integrin 7 to form the E-cadherin ␣ ␤ the lumenal surface of the airway epithelium (10, 11). Isolation of ligand E 7, is a more discriminating marker for the two major these epithelial airway DC by microdissection showed that they lung DC subsets. The basolateral expression of E-cadherin by bronchial epithelial cells immediately explains the epithelial local- ␣ ␤ ϩ ϩ ization of integrin E 7 DC (CD103 DC) and this localization ϩ Department of Internal Medicine, University of Virginia Health Sciences Center, suggests that the CD103 DC type corresponds to the epithelial Charlottesville, VA 22908 DC reported earlier and is largely responsible for the presentation Received for publication July 26, 2006. Accepted for publication November 14, 2006. of airway Ag. Their high expression of tight junction proteins that The costs of publication of this article were defrayed in part by the payment of page allows the ready access of DC extensions into the airway lumen charges. This article must therefore be hereby marked advertisement in accordance ϩ with 18 U.S.C. Section 1734 solely to indicate this fact. further supports their Ag presentation role (14). CD103 DC have 1 This work was supported by U.S. Public Health Service Grants HL070065 and been described in intestinal lamina propria, mesenteric lymph HL065344 from the National Institutes of Health. nodes, and skin-draining lymph nodes, but not in spleen or skin 2 Address correspondence and reprint requests to Dr. Sun-sang J. Sung, Division of (15–18). In Ag- or pathogenic T cell-induced intestinal inflamma- Rheumatology and Immunology, Department of Internal Medicine, Box 800412, Uni- tion, the generation of gut-tropic T cells expressing CCR9 and versity of Virginia Health Sciences Center, Charlottesville, VA 22908. E-mail ad- ␣ ␤ dress: [email protected] integrin 4 7 (lymphocyte Peyer’s patch high endothelial venule 3 adhesion molecule 1 (LPAM-1)) requires the participation of Abbreviations used in this paper: AHR, airway hyperresponsiveness; 7-AAD, ϩ Ϫ ϩ ␣ ␤ ϩ 7-aminoactinomycin D; BLC, B cell chemoattractant; CD103 DC, integrin E 7 CD103 DC while CD103 DC failed to induce these T cells (17, DC; IP-10, IFN-␥-inducible protein of 10 kDa; DC, dendritic cell; I-TAC, IFN-␥- 18), thus differentiating the functional roles of CD103ϩ and inducible T cell ␣ chemoattractant; MDC, monocyte-derived chemokine; MIG, Ϫ monokine induced by IFN-␥; PDC, plasmacytoid DC; PF-4, platelet factor 4; PMN, CD103 DC in gut. However, for lung DC, besides tight junction neutrophil; poly(I:C), poly(deoxyinosinic-deoxycytidylic acid; TARC, thymus and proteins and adhesion molecules expression, few other surface activation-related chemokine; LPAM-1, lymphocyte Peyer’s patch high endothelial marker expression and functional differences between CD103ϩ venule adhesion molecule 1. and CD103ϪCD11bhigh DC have been documented. It is unclear Copyright © 2007 by The American Association of Immunologists, Inc. 0022-1767/07/$2.00 whether one subset can be derived from the other by activation www.jimmunol.org The Journal of Immunology 1883 with cytokines or other stimulants and the occurrence of compared to macrophages. The results indicate that lung CD11bhigh and CD103ϩ DC as distinct functional subset has not CD11bhigh DC is a major chemokine producer and dissociate their yet been established. One possibility is that these DC subsets are functions from CD103ϩ DC in their role in directing lung leuko- derived from I-AϪ precursor found in neonatal rats and in adult cyte trafficking during lung inflammation, although both DC sub- mice airways (19, 20), but this development has not been formally sets directed a Th2-biased response. shown in adults. Allergic diseases are characterized by Th2-mediated inflamma- tory responses (1, 2). In asthma, lung and peripheral blood T cells Materials and Methods are dominated by allergen-specific Th2 cells. The basis for this Materials biased-Th subset response has not been established but may be due TLR ligands were from InvivoGen and have been tested to be LPS free by to lung chemokine production that preferentially attract Th2 cells the manufacturer. All mAb were from eBioscience, except the following: ␤ (21–23), or the Th2-directing function of lung DC (24, 25). In anti-integrin 7 and SiglacF mAb were from BD Pharmingen; anti- spleen and lymph nodes, CD8ϩ DC have been found to be Th1- mPDCA-1mAb was from Miltenyi Biotec; and rabbit or goat anti- chemokine Ab were from PeproTech or R&D Systems. The polyclonal biased because of their ability to secrete high levels of IL-12 while ϩ anti-chemokine Ab were prepared by immunization with whole chemokine CD11b DC are Th2-directing (26). However, no Th1-directing proteins and affinity purified. Alexa Fluor-conjugated secondary Abs were ϩ CD8 DC have been found in lungs (14). This absence may in from Molecular Probes. Collagenase D and DNase I were from Roche. itself be the reason for the lung Th2-biased priming. However, Hyaluronidase was from Sigma-Aldrich. TaqDNA polymerase was from because Th subset response studies thus far have used bulk New England Biolabs. ϩ ϩ CD11c lung cells composed of CD11bhigh DC, CD103 DC, and Downloaded from macrophages, the DC subset functions in Th2 development have DC isolation and analyses by flow cytometer not been examined individually and need to be established.