DOCSLIB.ORG

Signaling and IL-6 Release 14/16 Α Pertussis Toxin-Insensitive G THP-1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Signaling and IL-6 Release 14/16 Α Pertussis Toxin-Insensitive G THP-1 CCR1-Mediated STAT3 Tyrosine Phosphorylation and CXCL8 Expression in THP-1 Macrophage-like Cells Involve Pertussis Toxin-Insensitive G α14/16 This information is current as Signaling and IL-6 Release of September 25, 2021. Maggie M. K. Lee, Ricky K. S. Chui, Issan Y. S. Tam, Alaster H. Y. Lau and Yung H. Wong J Immunol published online 2 November 2012 http://www.jimmunol.org/content/early/2012/10/31/jimmun Downloaded from ol.1103359 Supplementary http://www.jimmunol.org/content/suppl/2012/11/02/jimmunol.110335 http://www.jimmunol.org/ Material 9.DC1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 25, 2021 • Fast Publication! 4 weeks from acceptance to publication *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 © 2012 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published November 2, 2012, doi:10.4049/jimmunol.1103359 The Journal of Immunology CCR1-Mediated STAT3 Tyrosine Phosphorylation and CXCL8 Expression in THP-1 Macrophage-like Cells Involve Pertussis Toxin-Insensitive Ga14/16 Signaling and IL-6 Release Maggie M. K. Lee,*,† Ricky K. S. Chui,*,† Issan Y. S. Tam,‡ Alaster H. Y. Lau,‡ and Yung H. Wong*,†,x Agonists of CCR1 contribute to hypersensitivity reactions and atherosclerotic lesions, possibly via the regulation of the transcrip- tion factor STAT3. CCR1 was demonstrated to use pertussis toxin-insensitive Ga14/16 to stimulate phospholipase Cb and NF-kB, whereas both Ga14 and Ga16 are also capable of activating STAT3. The coexpression of CCR1 and Ga14/16 in human THP-1 macrophage-like cells suggests that CCR1 may use Ga14/16 to induce STAT3 activation. In this study, we demonstrated that 705 727 a CCR1 agonist, leukotactin-1 (CCL15), could indeed stimulate STAT3 Tyr and Ser phosphorylation via pertussis toxin- Downloaded from insensitive G proteins in PMA-differentiated THP-1 cells, human erythroleukemia cells, and HEK293 cells overexpressing CCR1 705 727 and Ga14/16. The STAT3 Tyr and Ser phosphorylations were independent of each other and temporally distinct. Subcellular fractionation and confocal microscopy illustrated that Tyr705-phosphorylated STAT3 translocated to the nucleus, whereas Ser727- phosphorylated STAT3 was retained in the cytosol after CCR1/Ga14 activation. CCL15 was capable of inducing IL-6 and IL-8 (CXCL8) production in both THP-1 macrophage-like cells and HEK293 cells overexpressing CCR1 and Ga14/16. Neutralizing Ab 705 to IL-6 inhibited CCL15-mediated STAT3 Tyr phosphorylation, whereas inhibition of STAT3 activity abolished CCL15- http://www.jimmunol.org/ activated CXCL8 release. The ability of CCR1 to signal through Ga14/16 provides a linkage for CCL15 to regulate IL-6/ STAT3–signaling cascades, leading to expression of CXCL8, a cytokine that is involved in inflammation and the rupture of atherosclerotic plaque. The Journal of Immunology, 2012, 189: 000–000. hemokines are a large family of low-molecular-weight mature cells. CCR1 knockout mice are defective in the trafficking cytokines that are characterized by their ability to direct and proliferation of myeloid progenitor cells (4). the migration of leukocytes from the bloodstream to sites of Hematopoiesis and angiogenesis require transcriptional activa- C by guest on September 25, 2021 inflammation (1). Leukotactin-1 (CCL15) belongs to the CC sub- tion, which can be mediated by STAT3. STAT3 is involved in T cell family, one of four chemokine groups (CXC, CC, C, and CX3C), as proliferation induced by IL-6 (11), c-Kit–mediated stem cell factor defined by their primary structures. CCL15 exerts its effect mainly (SCF)-independent proliferation in human leukemia cells (12), via CCR1 (2, 3), which is a G protein-coupled receptor. In addition and vessel formation triggered by GM-CSF (13). Deletion of to mediating chemotaxis, CCL15 and CCR1 were shown to regulate STAT3 is embryonic lethal (14). In addition, STAT3 acts as a hematopoiesis (4), angiogenesis (5), mast cell activation (6), and negative regulator of inflammatory responses in hematopoietic inflammatory diseases, including atherosclerosis (7, 8). CCR1 is cells. Tissue-specific deletion of STAT3 in macrophages enhances expressed in myeloid progenitor cells (9) and endothelial cells (10), the production of inflammatory cytokines (15), whereas disruption both of which are capable of proliferating and differentiating into of STAT3 during hematopoiesis leads to severe inflammatory bowel disease (16). In addition, expression of RANTES/CCL5 (a CCR1 agonist) is regulated, in part, by a transcription complex of STAT3 and NF-kB (17). *Division of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China; †Biotechnology Research Institute, Hong Although chemokine receptors are typically characterized as Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, Gi-coupled receptors, there is substantial evidence to suggest that ‡ China; School of Biomedical Sciences, Faculty of Medicine, Chinese University of chemokines may be able to stimulate STAT3 activity through Hong Kong, Hong Kong, China; and xState Key Laboratory of Molecular Neurosci- ence, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, pertussis toxin (PTX)-insensitive G proteins. CCR1 agonists were Hong Kong, China found to induce gene expression of the STAT-inducible proto- Received for publication November 23, 2011. Accepted for publication September oncogene, c-fos (18). c-Fos expression and transcriptional acti- 28, 2012. vation is induced upon Ga16 activation (19), and constitutively This work was supported in part by grants from the Research Grants Council of Hong active mutants of Ga14 and Ga16 were demonstrated to enhance Kong (HKUST 1/06C, 663108, and AoE/B-15/01) and the Hong Kong Jockey Club. the activity of STAT3 in cotransfection systems (20, 21). The Address correspondence and reprint requests to Prof. Yung H. Wong, Division of Life coexistence of Ga and CCR1, as well as their demonstrated Science, Hong Kong University of Science and Technology, Clear Water Bay, Kow- 14/16 loon, Hong Kong, China. E-mail address: [email protected] functional coupling in THP-1 macrophage-like cells (2, 22–26), The online version of this article contains supplemental material. suggests that CCR1 may use Ga14/16 to stimulate STAT3. There is Abbreviations used in this article: bFGF, basic fibroblast growth factor; EGF, epider- increasing evidence to support a role for STAT3 activation in mal growth factor; HEL, human erythroleukemia; MMP, matrix metalloproteinase; CCR1-mediated cellular responses. In human macrophages and PLCb, phospholipase Cb; PTX, pertussis toxin; SCF, stem cell factor; siRNA, small macrophage-derived foam cells, CCL15 promotes the release of interfering RNA. matrix metalloproteinase (MMP)-9 (27), which is implicated in Copyright Ó 2012 by The American Association of Immunologists, Inc. 0022-1767/12/$16.00 the progression of atherosclerosis and whose expression is regu- www.jimmunol.org/cgi/doi/10.4049/jimmunol.1103359 2 INDUCTION OF CXCL8 BY CCR1/Ga14/16 REQUIRES IL-6/STAT3 lated by STAT3 (28). Moreover, CCR1 was shown to mediate IL-6 Cells were transiently transfected with cDNAs encoding p-STAT3–TA– production in marrow stromal cells upon stimulation by human Luc, STAT3 or its phosphorylation-resistant mutants (500 ng) or wild-type or constitutively active forms of Ga (625 ng) using Lipofectamine Plus myeloma cells (29), whereas MCP-1/CCL2 (a CCR2 agonist) reagent. enhances IL-6 production in fibroblast-like synoviocytes from patients with rheumatoid arthritis, and the response is mediated, in Luciferase reporter assay part, via PTX-insensitive G proteins (30). In addition, in THP-1 After 24 h of transfection, cells were serum starved with 100 ng/ml PTX for macrophage-like cells, lipoprotein (31) and human placenta 4 h and then stimulated in the absence or presence of 10 nM CCL15 for extracts (32) were demonstrated to promote the expression of another 24 h. Subsequently, the assay medium (culture medium without FBS) was removed and replaced by 150 ml lysis buffer provided in the IL-8 (CXCL8), which is capable of inducing MMP expression Luciferase Reporter Gene Assay Kit (Roche Applied Science, Penzberg, (33). Interestingly, both IL-6 (34) and CXCL8 (35) are STAT3- Upper Bavaria, Germany). The 6-well plate was shaken at 4˚C for 30 min, regulated cytokines, and we recently demonstrated that activa- and 25 ml lysate was transferred to white 96-well microplates designed for tion of Ga can lead to STAT3 activation and upregulation of luminescent work (Nunc, Roskilde, Denmark). An additional 25 ml lysis 16 buffer and 25 ml luciferin substrate were added to each well to initiate the CXCL8 via IL-6 autocrine signaling in HEK293 and human Jurkat reaction. Luciferase activity was determined using a microplate lumino- T cells (36). Given the importance of STAT3 in hematopoiesis and meter LB96V (EG&G Berthold, Bad Wildbad, Germany) (20, 21). its purported involvement in inflammatory diseases, we explored Assay for STAT3 phosphorylation whether CCR1 can indeed induce STAT3 phosphorylation and the production of IL-6 and CXCL8 through Ga -mediated sig- HEL cells, as well as THP-1 and U-937 macrophage-like cells, were seeded 14/16 3 6 naling. Mapping of such a pathway will help to elucidate the in- at 1 10 cells in assay medium (culture medium containing 0.1% BSA instead of FBS) and cultured or not with 100 ng/ml PTX for 16 h.
Load more

Recommended publications
  • Cellular and Plasma Proteomic Determinants of COVID-19 and Non-COVID-19 Pulmonary Diseases Relative to Healthy Aging
    RESOURCE https://doi.org/10.1038/s43587-021-00067-x Cellular and plasma proteomic determinants of COVID-19 and non-COVID-19 pulmonary diseases relative to healthy aging Laura Arthur1,8, Ekaterina Esaulova 1,8, Denis A. Mogilenko 1, Petr Tsurinov1,2, Samantha Burdess1, Anwesha Laha1, Rachel Presti 3, Brian Goetz4, Mark A. Watson1, Charles W. Goss5, Christina A. Gurnett6, Philip A. Mudd 7, Courtney Beers4, Jane A. O’Halloran3 and Maxim N. Artyomov1 ✉ We examine the cellular and soluble determinants of coronavirus disease 2019 (COVID-19) relative to aging by performing mass cytometry in parallel with clinical blood testing and plasma proteomic profiling of ~4,700 proteins from 71 individuals with pul- monary disease and 148 healthy donors (25–80 years old). Distinct cell populations were associated with age (GZMK+CD8+ T cells and CD25low CD4+ T cells) and with COVID-19 (TBET−EOMES− CD4+ T cells, HLA-DR+CD38+ CD8+ T cells and CD27+CD38+ B cells). A unique population of TBET+EOMES+ CD4+ T cells was associated with individuals with COVID-19 who experienced moderate, rather than severe or lethal, disease. Disease severity correlated with blood creatinine and urea nitrogen levels. Proteomics revealed a major impact of age on the disease-associated plasma signatures and highlighted the divergent contri- bution of hepatocyte and muscle secretomes to COVID-19 plasma proteins. Aging plasma was enriched in matrisome proteins and heart/aorta smooth muscle cell-specific proteins. These findings reveal age-specific and disease-specific changes associ- ated with COVID-19, and potential soluble mediators of the physiological impact of COVID-19.
    [Show full text]
  • Anti-OX40 Antibody Directly Enhances the Function of Tumor-Reactive CD8+ T Cells
    Author Manuscript Published OnlineFirst on August 1, 2019; DOI: 10.1158/1078-0432.CCR-19-1259 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. 1 Anti-OX40 antibody directly enhances the function of tumor-reactive CD8+ T cells and synergizes with PI3Kβ inhibition in PTEN loss melanoma Weiyi Peng1,5*, Leila J. Williams1, Chunyu Xu1,5, Brenda Melendez1, Jodi A. McKenzie1,6, Yuan Chen1, Heather Jackson2, Kui S. Voo3, Rina M. Mbofung1,7,, Sara E. Leahey1, Jian Wang4, Greg Lizee1, Hussein A. Tawbi1, Michael A. Davies1, Axel Hoos2, James Smothers2, Roopa Srinivasan2, Elaine Paul2, Niranjan Yanamandra2* and Patrick Hwu1* 1Department of Melanoma Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX. 2Oncology R&D, Immuno-Oncology and Combinations RU, GlaxoSmithKline, 1250 S. Collegeville Rd, Collegeville, PA 19426, United States 3Oncology Research for Biologics and Immunotherapy Translation Platform, The University of Texas MD Anderson Cancer Center, Houston, TX. 4Department of Biostatistics, The University of Texas MD Anderson Cancer Center, Houston, TX. 5Present address: Department of Biology and Biochemistry, University of Houston, Houston, TX. 6Present address: Eisai Inc., Woodcliff Lake, NJ. 7Present address: Merck Research Laboratories, Palo Alto, CA. Running Title: OX40 agonist-based cancer immunotherapy Keywords: OX40, PI3K, cancer immunotherapy Downloaded from clincancerres.aacrjournals.org on September 25, 2021. © 2019 American Association for Cancer Research. Author Manuscript Published OnlineFirst on August 1, 2019; DOI: 10.1158/1078-0432.CCR-19-1259 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. 2 *Corresponding Authors: Patrick Hwu, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030.
    [Show full text]
  • Neutrophil Chemoattractant Receptors in Health and Disease: Double-Edged Swords
    Cellular & Molecular Immunology www.nature.com/cmi REVIEW ARTICLE Neutrophil chemoattractant receptors in health and disease: double-edged swords Mieke Metzemaekers1, Mieke Gouwy1 and Paul Proost 1 Neutrophils are frontline cells of the innate immune system. These effector leukocytes are equipped with intriguing antimicrobial machinery and consequently display high cytotoxic potential. Accurate neutrophil recruitment is essential to combat microbes and to restore homeostasis, for inflammation modulation and resolution, wound healing and tissue repair. After fulfilling the appropriate effector functions, however, dampening neutrophil activation and infiltration is crucial to prevent damage to the host. In humans, chemoattractant molecules can be categorized into four biochemical families, i.e., chemotactic lipids, formyl peptides, complement anaphylatoxins and chemokines. They are critically involved in the tight regulation of neutrophil bone marrow storage and egress and in spatial and temporal neutrophil trafficking between organs. Chemoattractants function by activating dedicated heptahelical G protein-coupled receptors (GPCRs). In addition, emerging evidence suggests an important role for atypical chemoattractant receptors (ACKRs) that do not couple to G proteins in fine-tuning neutrophil migratory and functional responses. The expression levels of chemoattractant receptors are dependent on the level of neutrophil maturation and state of activation, with a pivotal modulatory role for the (inflammatory) environment. Here, we provide an overview
    [Show full text]
  • Follicular Thyroid Carcinoma but Not Adenoma Recruits Tumor-Associated
    Huang et al. BMC Cancer (2016) 16:98 DOI 10.1186/s12885-016-2114-7 RESEARCH ARTICLE Open Access Follicular thyroid carcinoma but not adenoma recruits tumor-associated macrophages by releasing CCL15 Feng-Jiao Huang1†, Xiao-Yi Zhou1†, Lei Ye1*, Xiao-Chun Fei2, Shu Wang1,3, Weiqing Wang1 and Guang Ning1,3 Abstract Background: The differential diagnosis of follicular thyroid carcinoma (FTC) and follicular adenoma (FA) before surgery is a clinical challenge. Many efforts have been made but most focusing on tumor cells, while the roles of tumor associated macrophages (TAMs) remained unclear in FTC. Here we analyzed the differences between TAMs in FTC and those in FA. Methods: We first analyzed the density of TAMs by CD68 immunostaining in 59 histologically confirmed FTCs and 47 FAs. Cytokines produced by FTC and FA were profiled using antibody array, and validated by quantitative PCR. Chemotaxis of monocyte THP-1 was induced by condition medium of FTC cell lines (FTC133 and WRO82-1) with and without anti-CCL15 neutralizing antibody. Finally, we analyzed CCL15 protein level in FTC and FA by immunohistochemistry. Results: The average density of CD68+ cells was 9.5 ± 5.4/field in FTC, significantly higher than that in FA (4.9 ± 3.4/field, p < 0.001). Subsequently profiling showed that CCL15 was the most abundant chemokine in FTC compared with FA. CCL15 mRNA in FTC was 51.4-folds of that in FA. CM of FTC cell lines induced THP-1 cell chemotaxis by 33 ~ 77 %, and anti-CCL15 neutralizing antibody reduced THP-1 cell migration in a dose-dependent manner.
    [Show full text]
  • Part One Fundamentals of Chemokines and Chemokine Receptors
    Part One Fundamentals of Chemokines and Chemokine Receptors Chemokine Receptors as Drug Targets. Edited by Martine J. Smit, Sergio A. Lira, and Rob Leurs Copyright Ó 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim ISBN: 978-3-527-32118-6 j3 1 Structural Aspects of Chemokines and their Interactions with Receptors and Glycosaminoglycans Amanda E. I. Proudfoot, India Severin, Damon Hamel, and Tracy M. Handel 1.1 Introduction Chemokines are a large subfamily of cytokines (50 in humans) that can be distinguished from other cytokines due to several features. They share a common biological activity, which is the control of the directional migration of leukocytes, hence their name, chemoattractant cytokines. They are all small proteins (approx. 8 kDa) that are highly basic, with two exceptions (MIP-1a, MIP-1b). Also, they have a highly conserved monomeric fold, constrained by 1–3 disulfides which are formed from a conserved pattern of cysteine residues (the majority of chemokines have four cysteines). The pattern of cysteine residues is used as the basis of their division into subclasses and for their nomenclature. The first class, referred to as CXC or a-chemokines, have a single residue between the first N-terminal Cys residues, whereas in the CC class, or b-chemokines, these two Cys residues are adjacent. While most chemokines have two disulfides, the CC subclass also has three members that contain three. Subsequent to the CC and CXC families, two fi additional subclasses were identi ed, the CX3C subclass [1, 2], which has three amino acids separating the N-terminal Cys pair, and the C subclass, which has a single disulfide.
    [Show full text]
  • Th22 Cells Represent a Distinct Human T Cell Subset Involved in Epidermal Immunity and Remodeling
    Th22 cells represent a distinct human T cell subset involved in epidermal immunity and remodeling Stefanie Eyerich, … , Carsten B. Schmidt-Weber, Andrea Cavani J Clin Invest. 2009;119(12):3573-3585. https://doi.org/10.1172/JCI40202. Research Article Immunology Th subsets are defined according to their production of lineage-indicating cytokines and functions. In this study, we have identified a subset of human Th cells that infiltrates the epidermis in individuals with inflammatory skin disorders and is characterized by the secretion of IL-22 and TNF-α, but not IFN-γ, IL-4, or IL-17. In analogy to the Th17 subset, cells with this cytokine profile have been named the Th22 subset. Th22 clones derived from patients with psoriasis were stable in culture and exhibited a transcriptome profile clearly separate from those of Th1, Th2, and Th17 cells; it included genes encoding proteins involved in tissue remodeling, such as FGFs, and chemokines involved in angiogenesis and fibrosis. Primary human keratinocytes exposed to Th22 supernatants expressed a transcriptome response profile that included genes involved in innate immune pathways and the induction and modulation of adaptive immunity. These proinflammatory Th22 responses were synergistically dependent on IL-22 and TNF-α. Furthermore, Th22 supernatants enhanced wound healing in an in vitro injury model, which was exclusively dependent on IL-22. In conclusion, the human Th22 subset may represent a separate T cell subset with a distinct identity with respect to gene expression and function, present within the epidermal layer in inflammatory skin diseases. Future strategies directed against the Th22 subset may be of value in chronic inflammatory skin disorders.
    [Show full text]
  • Development and Validation of a Protein-Based Risk Score for Cardiovascular Outcomes Among Patients with Stable Coronary Heart Disease
    Supplementary Online Content Ganz P, Heidecker B, Hveem K, et al. Development and validation of a protein-based risk score for cardiovascular outcomes among patients with stable coronary heart disease. JAMA. doi: 10.1001/jama.2016.5951 eTable 1. List of 1130 Proteins Measured by Somalogic’s Modified Aptamer-Based Proteomic Assay eTable 2. Coefficients for Weibull Recalibration Model Applied to 9-Protein Model eFigure 1. Median Protein Levels in Derivation and Validation Cohort eTable 3. Coefficients for the Recalibration Model Applied to Refit Framingham eFigure 2. Calibration Plots for the Refit Framingham Model eTable 4. List of 200 Proteins Associated With the Risk of MI, Stroke, Heart Failure, and Death eFigure 3. Hazard Ratios of Lasso Selected Proteins for Primary End Point of MI, Stroke, Heart Failure, and Death eFigure 4. 9-Protein Prognostic Model Hazard Ratios Adjusted for Framingham Variables eFigure 5. 9-Protein Risk Scores by Event Type This supplementary material has been provided by the authors to give readers additional information about their work. Downloaded From: https://jamanetwork.com/ on 10/02/2021 Supplemental Material Table of Contents 1 Study Design and Data Processing ......................................................................................................... 3 2 Table of 1130 Proteins Measured .......................................................................................................... 4 3 Variable Selection and Statistical Modeling ........................................................................................
    [Show full text]
  • Human CCL15/MIP-1Δ Antibody
    Human CCL15/MIP-1δ Antibody Monoclonal Mouse IgG1 Clone # 88119 Catalog Number: MAB3631 DESCRIPTION Species Reactivity Human Specificity Detects human CCL15/MIP­1δ in direct ELISAs and Western blots. In direct ELISAs, no cross­reactivity with recombinant human CCL1, 2, 3, 4, 5, 7, 8, 11, 13, 14, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, recombinant mouse CCL1, 2, 3, 4, 5, 6, 7, 9, 11, 12, 17, 19, 20, 21, 22, 24, 25, or recombinant rat CCL20 is observed. Source Monoclonal Mouse IgG1 Clone # 88119 Purification Protein A or G purified from ascites Immunogen E. coli­derived recombinant human CCL15/MIP­1δ Ser46­Ile113 Accession # Q16663 Endotoxin Level <0.10 EU per 1 μg of the antibody by the LAL method. Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (­SP) is supplied either lyophilized or as a 0.2 μm filtered solution in PBS. APPLICATIONS Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website. Recommended Sample Concentration Western Blot 1 µg/mL Recombinant Human CCL15/MIP­1δ 92 aa (Catalog # 363­MG) Neutralization Measured by its ability to neutralize CCL15/MIP­1δ­induced chemotaxis in the BaF3 mouse pro­B cell line transfected with human CCR1. The Neutralization Dose (ND50) is typically 5­20 µg/mL in the presence of 1 ng/mL Recombinant Human CCL15/MIP­1δ 68 aa. DATA Neutralization Chemotaxis Induced by CCL15/MIP­1δ and Neutral­ ization by Human CCL15/ MIP­1δ Antibody.
    [Show full text]
  • CCL20/CCR6 Signaling Regulates Bone Mass Accrual in Mice
    ORIGINAL ARTICLE JBMR CCL20/CCR6 Signaling Regulates Bone Mass Accrual in Mice Michele Doucet, Swaathi Jayaraman, Emily Swenson, Brittany Tusing, Kristy L Weber,Ã and Scott L Kominsky Department of Orthopaedic Surgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA ABSTRACT CCL20 is a member of the macrophage inflammatory protein family and is reported to signal monogamously through the receptor CCR6. Although studies have identified the genomic locations of both Ccl20 and Ccr6 as regions important for bone quality, the role of CCL20/CCR6 signaling in regulating bone mass is unknown. By micro–computed tomography (mCT) and histomorphometric analysis, we show that global loss of Ccr6 in mice significantly decreases trabecular bone mass coincident with reduced osteoblast numbers. Notably, CCL20 and CCR6 were co-expressed in osteoblast progenitors and levels increased during osteoblast differentiation, indicating the potential of CCL20/CCR6 signaling to influence osteoblasts through both autocrine and paracrine actions. With respect to autocrine effects, CCR6 was found to act as a functional G protein–coupled receptor in osteoblasts and although its loss did not appear to affect the number or proliferation rate of osteoblast progenitors, differentiation was significantly inhibited as evidenced by delays in osteoblast marker gene expression, alkaline phosphatase activity, and mineralization. In addition, CCL20 promoted osteoblast survival concordant with activation of the PI3K-AKT pathway. Beyond these potential autocrine effects, osteoblast-derived CCL20 stimulated the recruitment of macrophages and T cells, known facilitators of osteoblast differentiation and survival. Finally, we generated mice harboring a global deletion of Ccl20 and found that Ccl20-/- mice exhibit a reduction in bone mass similar to that observed in Ccr6-/- mice, confirming that this phenomenon is regulated by CCL20 rather than alternate CCR6 ligands.
    [Show full text]
  • STAT6-Dependent Fashion CCL23 Expression Is Induced by IL-4 in A
    CCL23 Expression Is Induced by IL-4 in a STAT6-Dependent Fashion Hermann Novak, Anke Müller, Nathalie Harrer, Claudia Günther, Jose M. Carballido and Maximilian Woisetschläger This information is current as of September 28, 2021. J Immunol 2007; 178:4335-4341; ; doi: 10.4049/jimmunol.178.7.4335 http://www.jimmunol.org/content/178/7/4335 Downloaded from References This article cites 47 articles, 20 of which you can access for free at: http://www.jimmunol.org/content/178/7/4335.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • 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 28, 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 © 2007 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology CCL23 Expression Is Induced by IL-4 in a STAT6-Dependent Fashion Hermann Novak, Anke Mu¨ller, Nathalie Harrer, Claudia Gu¨nther, Jose M. Carballido, and Maximilian Woisetschla¨ger1 The chemokine CCL23 is primarily expressed in cells of the myeloid lineage but little information about its regulation is available.
    [Show full text]
  • 1599.Full.Pdf
    Quantum Proteolytic Activation of Chemokine CCL15 by Neutrophil Granulocytes Modulates Mononuclear Cell Adhesiveness This information is current as of September 30, 2021. Rudolf Richter, Roxana Bistrian, Sylvia Escher, Wolf-Georg Forssmann, Jalal Vakili, Reinhard Henschler, Nikolaj Spodsberg, Adjoa Frimpong-Boateng and Ulf Forssmann J Immunol 2005; 175:1599-1608; ; doi: 10.4049/jimmunol.175.3.1599 Downloaded from http://www.jimmunol.org/content/175/3/1599 References This article cites 91 articles, 34 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/175/3/1599.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 30, 2021 • Fast Publication! 4 weeks from acceptance to publication *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 © 2005 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Quantum Proteolytic Activation of Chemokine CCL15 by Neutrophil Granulocytes Modulates Mononuclear Cell Adhesiveness1 Rudolf Richter,2* Roxana Bistrian,† Sylvia Escher,* Wolf-Georg Forssmann,* Jalal Vakili,‡ Reinhard Henschler,† Nikolaj Spodsberg,§ Adjoa Frimpong-Boateng,* and Ulf Forssmann* Monocyte infiltration into inflammatory sites is generally preceded by neutrophils.
    [Show full text]
  • Chapter 7 Autocrine Regulation of Re- Epithelialization After Wounding by Chemokine Receptors CCR1, CCR10, CXCR1, CXCR2 and CXCR3
    Chapter 7 Autocrine regulation of re- epithelialization after wounding by chemokine receptors CCR1, CCR10, CXCR1, CXCR2 and CXCR3 Kim Kroeze Mireille Boink Shakun Sampat - Sardjoepersad Taco Waaijman Rik Scheper Susan Gibbs Journal of Investigative Dermatology, 2011;132:215-225 Autocrine regulation of re-epithelialization after wounding by chemokine receptors CCR1, CCR10, CXCR1, CXCR2 and CXCR3 ABStract This study identifies chemokine receptors involved in an autocrine regulation of re-epitheli- alization after skin tissue damage. We determined which receptors, from a panel of thirteen, are expressed in healthy human epidermis and which mono-specific chemokine ligands, se- creted by keratinocytes, were able to stimulate migration and proliferation. A reconstructed epidermis cryo-(freeze) wound model was used to assess chemokine secretion after wound- ing and the effect of pertussis toxin (chemokine receptor blocker) on re-epithelialization and differentiation. Chemokine receptors CCR1, CCR3, CCR4, CCR6, CCR10, CXCR1, CXCR2, CXCR3 and CXCR4 were expressed in epidermis. No expression of CCR2, CCR5, CCR7 and CCR8 was observed by either immunostaining or flow cytometry. Five chemokine receptors (CCR1, CCR10, CXCR1, CXCR2, CXCR3) were identified whose corresponding mono-specific ligands (CCL14, CCL27, CXCL8, CXCL1, CXCL10 respectively) were not only able to stimulate keratinocyte migration and/or proliferation but were also secreted by keratinocytes after introducing cryo-wounds into epidermal equivalents. Blocking of receptor-ligand interac- tions with pertussis toxin delayed re-epithelialization but did not influence differentiation (as assessed by formation of basal layer, spinous layer, granular layer and stratum corneum) after cryo-wounding. Taken together, these results confirm that an autocrine positive feedback loop of epithelialization exists in order to stimulate wound closure after skin injury.
    [Show full text]