Modulation of Dendritic Cell Differentiation and Maturation by Decoy Receptor 3 Tsui-Ling Hsu, Yung-Chi Chang, Siu-Ju Chen, Yong-Jun Liu, Allen W. Chiu, Chung-Ching Chio, Lieping Chen and This information is current as Shie-Liang Hsieh of September 25, 2021. J Immunol 2002; 168:4846-4853; ; doi: 10.4049/jimmunol.168.10.4846 http://www.jimmunol.org/content/168/10/4846 Downloaded from References This article cites 39 articles, 21 of which you can access for free at: http://www.jimmunol.org/content/168/10/4846.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 *average by guest on September 25, 2021 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 © 2002 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Modulation of Dendritic Cell Differentiation and Maturation by Decoy Receptor 31 Tsui-Ling Hsu,* Yung-Chi Chang,* Siu-Ju Chen,* Yong-Jun Liu,§ Allen W. Chiu,† Chung-Ching Chio,† Lieping Chen,‡ and Shie-Liang Hsieh2* Decoy receptor 3 (DcR3), a soluble receptor belonging to the TNFR superfamily, is a receptor for both Fas ligand (FasL) and LIGHT. It has been demonstrated that DcR3 is up-regulated in lung and colon cancers, thus promoting tumor growth by neutralizing the cytotoxic effects of FasL and LIGHT. In this study, we found that DcR3.Fc profoundly modulated dendritic cell differentiation and maturation from CD14؉ monocytes, including the up-regulation of CD86/B7.2, and the down-regulation of -CD40, CD54/ICAM-1, CD80/B7.1, CD1a, and HLA-DR. Moreover, DcR3-treated dendritic cells suppressed CD4؉ T cell prolif ؉ ؉ eration in an allogeneic MLR and up-regulated IL-4 secretion of CD4 CD45RA T cells. This suggests that DcR3.Fc may act not Downloaded from only as a decoy receptor to FasL and LIGHT, but also as an effector molecule to skew T cell response to the Th2 phenotype. The Journal of Immunology, 2002, 168: 4846–4853. ecoy receptor 3 (DcR3),3 also known as TR6 or M68, is surveillance during lymphomagenesis, or that virus-infected lym- a member of the TNFR superfamily and is a decoy re- phoma cells with DcR3 expression might be selected during mul- ceptor for Fas ligand (FasL) and LIGHT (homologous to tistep tumorigenesis (6). In addition, expression of DcR3 can be D http://www.jimmunol.org/ lymphotoxins, shows inducible expression, and competes with detected in malignant glioma cells as well as in human glioblas- HSV glycoprotein D for herpesvirus entry mediator, a receptor tomas, and its expression correlates with the grade of malignancy expressed by T lymphocytes) (1–3). Like osteoprotegerin (4), a (7). Besides tumor cell patients, the DcR3 gene is also overexpressed member of the TNFR superfamily, DcR3 lacks a transmembrane in silicosis or systemic lupus erythematosus patients (8). Because domain and is regarded as a secreted, rather than a membrane- LIGHT is expressed in dendritic cells (DCs) and acts as a costimu- associated, molecule. Moreover, DcR3 can apparently neutralize latory factor essential for priming T cell responses (9–11), we ques- the biological effects of FasL and LIGHT by inhibiting the FasL- tioned whether DcR3 could suppress immunity by interfering with the Fas interaction (1) or by inhibiting LIGHT binding to both the maturation and differentiation of DCs. lymphotoxin (LT)-␤ receptor (LT␤R) and the herpes virus entry by guest on September 25, 2021 Growing evidence has demonstrated that members of the TNF mediator (2, 5). DcR3 gene expression is increased in malignant superfamily transduce signals after engagement with their recep- tissue (1) and DcR3 protein is overexpressed in human adenocar- tors (12–21). In our recent study, we further demonstrated that cinomas of the esophagus, stomach, colon, and rectum (3). More- cross-linking of TNF-related activation-induced cytokine by im- over, DcR3 protein was overexpressed in a substantial number of mobilized soluble receptor activator of NF-␬B.Fc fusion protein tumors in which gene amplification could not be detected (3). A recent study further demonstrated that DcR3 is amplified and over- activated p38 mitogen-activated protein kinase and enhanced ␥ expressed in virus (EBV or human T cell leukemia virus-I)-asso- IFN- secretion via reverse signaling through TNF-related activa- ciated lymphomas (6). These results suggest that EBV and human tion-induced cytokine (22), and cross-linking of TNF-related ap- ␥ T cell leukemia virus-I may use DcR3 to escape from immune optosis-inducing ligand, enhanced proliferation, and IFN- secre- tion of T cells (23). However, even though “reverse signaling” could be triggered by *Department of Microbiology and Immunology, and Immunology Research Center, National Yang-Ming University, Taipei, Taiwan; †Department of Surgery, Chi-Mei immobilized receptor.Fc or agonistic mAb, there is no evidence to Foundational Hospital, Tainan, Taiwan; ‡Department of Immunology, Mayo Gradu- demonstrate that the soluble receptor.Fc fusion protein can trigger § ate and Medical Schools, Mayo Clinic, Rochester, MN 55905; and Department of signaling and modulate cell function. In this study, we report that Immunobiology, DNAX Research Institute of Molecular and Cellular Biology, Palo ϩ Alto, CA 94304 soluble DcR3.Fc binds to CD14 monocytes and interferes with Received for publication October 22, 2001. Accepted for publication March 5, 2002. their differentiation and maturation into DCs. The expression of The costs of publication of this article were defrayed in part by the payment of page HLA-DR and other costimulatory molecules, such as CD40 and charges. This article must therefore be hereby marked advertisement in accordance CD80/B7.1, was suppressed. In contrast, the costimulatory mole- with 18 U.S.C. Section 1734 solely to indicate this fact. cule, CD86/B7.2, was up-regulated under the same conditions. 1 This work was mainly supported by Grant NHRI-CN-BP-8902S from the National Health Research Institute, Taiwan; National Sciences Council Grants NSC90-2320- Moreover, DcR3.Fc-treated DCs biased T cell differentiation to the B-010-109 and NSC90-2318-B-010-009-M51; and Grant GMYM 8902 from the Chi- Th2 phenotype in allogeneic MLR. Similar results were not ob- Mei Foundational Hospital (Tainan, Taiwan). served when Fas.Fc or LT␤R.Fc was used in place of DcR3.Fc. 2 Address correspondence and reprint requests to Dr. Shie-Liang Hsieh, Department Because DcR3.Fc fusion protein has been shown to have the sim- and Institute of Microbiology and Immunology, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan. E-mail address: [email protected] ilar binding affinity and specificity as that of DcR3 (24, 25), this 3 Abbreviations used in this paper: DcR3, decoy receptor 3; FasL, Fas ligand; LT, raises the argument that DcR3 produced by many human tumor lymphotoxin; LIGHT, homologous to LTs, shows inducible expression, and competes cells might have similar function to DcR3.Fc and could directly with HSV glycoprotein D for herpesvirus entry mediator, a receptor expressed by T lymphocytes; LT␤R, LT␤ receptor; DC, dendritic cell; sLIGHT, soluble LIGHT; suppress host anti-tumor immunity by altering DCs function and CD40L, CD40 ligand. skewing the immune response from Th1 to Th2. Copyright © 2002 by The American Association of Immunologists 0022-1767/02/$02.00 The Journal of Immunology 4847 Materials and Methods incubated at 4°C for 20 min with 50 ␮l UltraAvidin-PE (Leinco Technol- Production of receptor.Fc fusion proteins ogies, Ballwin, MO) or streptavidin-FITC (BD PharMingen) diluted (1/200) in FACS staining/washing buffer. The sources of mAbs are as LT␤R.Fc protein was produced as previously described (10). To generate follows: anti-CD1a-FITC (clone HI149; BD PharMingen), anti-CD11c-PE the DcR3.Fc, the open reading frame of the human DcR3 gene was isolated (B-ly6; BD PharMingen), anti-CD54 (clone 8.4A6; Ancell, Bayport, MN), by RT-PCR using the forward primer: 5Ј-GGAATTCAAGGACCAT anti-CD80-PE (clone L307.4; BD PharMingen), anti-CD83-FITC (clone GAGGGCGCTG-3Ј and the reverse primer: 5Ј-GGAATTCGTGCACA HB15e; BD PharMingen), anti-CD86-PE (clone 2331 FUN-1; BD PharM- GGGAGGAAGCGC-3Ј. The amplified product was ligated in-frame into ingen), anti-CD40-FITC (clone LOB716; Serotec, Oxford, U.K.), anti- the EcoRI-cut pUC19-IgG1-Fc vector containing the cDNA of the human HLA-DR-FITC (clone B-F1; Serotec), anti-CD14-PE mAb (clone IgG1 Fc. The fusion gene was then subcloned into the pBacPAK9 vector UCHM1; Serotec), biotin-conjugated anti-FasL (clone NOK-1; BD Phar- (Clontech Laboratories, Palo Alto, CA) and cotransfected with linearized Mingen), anti-TNF-␤/LT-␣ mAb (clone 9B9; Boehringer Mannheim, BacPAK6 DNA (Clontech Laboratories) into Sf21 cells. The supernatant Mannheim, Germany), and anti-DC-lysosome-associated membrane gly- from recombinant virus-infected Sf21 cells was filtered and purified on coprotein mAb (clone 104.G4; Immunotech, Marseille, France). protein A-Sepharose beads. The bound DcR3.Fc protein was then eluted with 0.1 M glycine buffer (pH 3.0) followed by dialysis against PBS. Immunoprecipitation of FasL and LIGHT Culture of CD14ϩ monocyte-derived DCs and preparation of COS7 cells were transfected with the pFLAG-FasL, pFLAG-LIGHT, or ϩ pFLAG-CMV2 vector by the calcium phosphate method.