title: The Notch Ligand DLL4 Defines a Capability of Human Dendritic Cells in Regulating Th1 and Th17 Differentiation.
abstract: Notch signaling regulates multiple helper CD4(+) T cell programs. We have recently demonstrated that dendritic cells (DCs) expressing the Notch ligand DLL4 are critical for eliciting alloreactive T cell responses and induction of graft-versus-host disease in mice. However, the human counterpart of murine DLL4(+) DCs has yet to be examined. We report the identification of human DLL4(+) DCs and their critical role in regulating Th1 and Th17 differentiation. CD1c(+) DCs and plasmacytoid DCs (pDCs) from the peripheral blood (PB) of healthy donors did not express DLL4. In contrast, patients undergoing allogeneic hematopoietic stem cell transplantation had a 16-fold more DLL4(+)CD1c(+) DCs than healthy donors. Upon activation of TLR signaling, healthy donor-derived CD1c(+) DCs dramatically upregulated DLL4, as did pDCs to a lesser extent. Activated DLL4(+) DCs were better able to promote Th1 and Th17 differentiation than unstimulated PB DCs. Blocking DLL4 using a neutralizing Ab decreased Notch signaling in T cells stimulated with DLL4(+) DCs, and it reduced the generation of Th1 and Th17 cells. Both NF-κB and STAT3 were crucial for inducing DLL4 in human DCs. Interestingly, STAT3 directly activated DLL4 transcription and inhibiting STAT3 alone was sufficient to reduce DLL4 in activated PB DCs. Thus, DLL4 is a unique functional molecule of human circulating DCs critical for directing Th1 and Th17 differentiation. These findings identify a pathway for therapeutic intervention for inflammatory disorders in humans, such as graft-versus-host disease after allogeneic hematopoietic stem cell transplantation, autoimmunity, and tumor immunity.
authors.affiliation: Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine, Shanghai 200231, China; Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140; Department of Microbiology and Immunology, Temple University, Philadelphia, PA 19140;
authors.forename: Lijun
authors.initials: L
authors.lastname: Meng
authors.affiliation: Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140; Pediatric Intensive Care Unit, Children's Hospital of Soochow University, Soochow University, Suzhou 215003, China;
authors.forename: Zhenjiang
authors.initials: Z
authors.lastname: Bai
authors.affiliation: Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140; Department of Microbiology and Immunology, Temple University, Philadelphia, PA 19140;
authors.forename: Shan
authors.initials: S
authors.lastname: He
authors.affiliation: Department of Pediatric Oncology, Fukushima Medical University, Fukushima 960-1295, Japan;
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Mochizuki
authors.affiliation: Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140;
authors.forename: Yongnian
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140; Department of Microbiology and Immunology, Temple University, Philadelphia, PA 19140;
authors.forename: Janaki
authors.identifier: http://orcid.org/0000-0001-7767-2700
authors.initials: J
authors.lastname: Purushe
authors.affiliation: Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140; Department of Microbiology and Immunology, Temple University, Philadelphia, PA 19140;
authors.forename: Hongxing
authors.initials: H
authors.lastname: Sun
authors.affiliation: Pediatric Intensive Care Unit, Children's Hospital of Soochow University, Soochow University, Suzhou 215003, China;
authors.forename: Jian
authors.identifier: http://orcid.org/0000-0003-2935-4964
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Immunology, Juntendo University School of Medicine, Tokyo 113-8421, Japan;
authors.forename: Hideo
authors.initials: H
authors.lastname: Yagita
authors.affiliation: Department of Medicine, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35294;
authors.forename: Shin
authors.initials: S
authors.lastname: Mineishi
authors.affiliation: Department of Hematology/Oncology, Fox Chase Cancer Center, Temple Health, Philadelphia, PA 19111;
authors.forename: Henry
authors.initials: H
authors.lastname: Fung
authors.affiliation: Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109;
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Yanik
authors.affiliation: Department of Medicine, Temple University School of Medicine, Philadelphia, PA 19148;
authors.forename: Roberto
authors.initials: R
authors.lastname: Caricchio
authors.affiliation: Flow Cytometry Core Facility, Temple University School of Medicine, Temple University, Philadelphia, PA 19148; and.
authors.forename: Xiaoxuan
authors.identifier: http://orcid.org/0000-0003-2823-4610
authors.initials: X
authors.lastname: Fan
authors.affiliation: Department of Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Crisalli
authors.affiliation: Department of Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104.
authors.forename: Elizabeth O
authors.identifier: http://orcid.org/0000-0002-1125-4060
authors.initials: EO
authors.lastname: Hexner
authors.affiliation: Department of Medicine, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104.
authors.forename: Ran
authors.identifier: http://orcid.org/0000-0003-2185-9546
authors.initials: R
authors.lastname: Reshef
authors.affiliation: Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine, Shanghai 200231, China; yi.zhang@temple.edu yyzhang@sibs.ac.cn.
authors.forename: Yanyun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140; Department of Microbiology and Immunology, Temple University, Philadelphia, PA 19140; yi.zhang@temple.edu yyzhang@sibs.ac.cn.
authors.forename: Yi
authors.identifier: http://orcid.org/0000-0002-3456-2728
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D064591:Allografts / Q000276:immunology; D015153:Blotting, Western; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D005434:Flow Cytometry; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology*; D007959:Lymphocyte Culture Test, Mixed; D060888:Real-Time Polymerase Chain Reaction; D018417:Th1 Cells / Q000166:cytology / Q000276:immunology*; D058504:Th17 Cells / Q000166:cytology / Q000276:immunology*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002135:Calcium-Binding Proteins; C424126:DLL4 protein, human; D036341:Intercellular Signaling Peptides and Proteins