Immobilized Heavy Chain-Hyaluronic Acid Polarizes Lipopolysaccharide-activated Macrophages toward M2 Phenotype*

Background: HC-HA is a unique anti-inflammatory matrix different from hyaluronic acid (HA). Results: Soluble HC-HA induces apoptosis of inflammatory neutrophils and macrophages, and immobilized HC-HA promotes M2 polarization upon LPS/TLR ligation while both enhancing macrophage phagocytosis. Conclusion: HC-HA exerts its anti-inflammatory action using multiple mechanisms. Significance: HC-HA is the first known matrix component to polarize M2b. Despite the known anti-inflammatory effect of amniotic membrane, its action mechanism remains largely unknown. HC-HA complex (HC-HA) purified from human amniotic membrane consists of high molecular weight hyaluronic acid (HA) covalently linked to the heavy chain (HC) 1 of inter-α-trypsin inhibitor. In this study, we show that soluble HC-HA also contained pentraxin 3 and induced the apoptosis of both formyl-Met-Leu-Phe or LPS-activated neutrophils and LPS-activated macrophages while not affecting the resting cells. This enhanced apoptosis was caused by the inhibition of cell adhesion, spreading, and proliferation caused by HC-HA binding of LPS-activated macrophages and preventing adhesion to the plastic surface. Preferentially, soluble HC-HA promoted phagocytosis of apoptotic neutrophils in resting macrophages, whereas immobilized HC-HA promoted phagocytosis in LPS-activated macrophages. Upon concomitant LPS stimulation, immobilized HC-HA but not HA polarized macrophages toward the M2 phenotype by down-regulating IRF5 protein and preventing its nuclear localization and by down-regulating IL-12, TNF-α, and NO synthase 2. Additionally, IL-10, TGF-β1, peroxisome proliferator-activated receptor γ, LIGHT (TNF superfamily 14), and sphingosine kinase-1 were up-regulated, and such M2 polarization was dependent on TLR ligation. Collectively, these data suggest that HC-HA is a unique matrix component different from HA and uses multiple mechanisms to suppress M1 while promoting M2 phenotype. This anti-inflammatory action of HC-HA is highly desirable to promote wound healing in diseases heightened by unsuccessful transition from M1 to M2 phenotypes.

[1]  P. Tak,et al.  The resolution of inflammation , 2012, Nature Reviews Immunology.

[2]  D. Underhill,et al.  Information processing during phagocytosis , 2012, Nature Reviews Immunology.

[3]  S. Okada,et al.  HIV-1 Proteins Preferentially Activate Anti-Inflammatory M2-Type Macrophages , 2012, The Journal of Immunology.

[4]  P. Muñoz-Cánoves,et al.  MKP-1 coordinates ordered macrophage-phenotype transitions essential for stem cell-dependent tissue repair , 2012, Cell cycle.

[5]  Alberto Mantovani,et al.  Macrophage plasticity and polarization: in vivo veritas. , 2012, The Journal of clinical investigation.

[6]  A. Heiligenhaus,et al.  Amniotic membrane induces peroxisome proliferator-activated receptor-γ positive alternatively activated macrophages. , 2012, Investigative ophthalmology & visual science.

[7]  S. Tseng,et al.  Constitutive Expression of Inter-α-inhibitor (IαI) Family Proteins and Tumor Necrosis Factor-stimulated Gene-6 (TSG-6) by Human Amniotic Membrane Epithelial and Stromal Cells Supporting Formation of the Heavy Chain-Hyaluronan (HC-HA) Complex , 2012, The Journal of Biological Chemistry.

[8]  T. Wynn,et al.  Protective and pathogenic functions of macrophage subsets , 2011, Nature Reviews Immunology.

[9]  D. Mosser,et al.  Regulatory macrophages: Setting the Threshold for Therapy , 2011, European journal of immunology.

[10]  T. Koh,et al.  Inflammation and wound healing: the role of the macrophage , 2011, Expert Reviews in Molecular Medicine.

[11]  S. Tseng,et al.  Inhibition of angiogenesis by HC·HA, a complex of hyaluronan and the heavy chain of inter-α-inhibitor, purified from human amniotic membrane. , 2011, Investigative ophthalmology & visual science.

[12]  H. Lockstone,et al.  IRF5 promotes inflammatory macrophage polarization and TH1-TH17 responses , 2011, Nature Immunology.

[13]  P. Pitha,et al.  Critical Role of IRF-5 in the Development of T helper 1 responses to Leishmania donovani infection , 2011, PLoS pathogens.

[14]  C. Haslett,et al.  Targeting granulocyte apoptosis: mechanisms, models, and therapies , 2010, Immunological reviews.

[15]  I. Udalova,et al.  IRF5 is required for late-phase TNF secretion by human dendritic cells. , 2010, Blood.

[16]  S. Gordon,et al.  Alternative activation of macrophages: mechanism and functions. , 2010, Immunity.

[17]  V. Hascall,et al.  Deficiency in the Serum-Derived Hyaluronan-Associated Protein-Hyaluronan Complex Enhances Airway Hyperresponsiveness in a Murine Model of Asthma , 2010, International Archives of Allergy and Immunology.

[18]  A. J. Day,et al.  The Angiogenic Inhibitor Long Pentraxin PTX3 Forms an Asymmetric Octamer with Two Binding Sites for FGF2* , 2010, The Journal of Biological Chemistry.

[19]  C. Haslett,et al.  The cyclin‐dependent kinase inhibitor R‐roscovitine down‐regulates Mcl‐1 to override pro‐inflammatory signalling and drive neutrophil apoptosis , 2010, European journal of immunology.

[20]  Oliver Soehnlein,et al.  Deficient CD40-TRAF6 signaling in leukocytes prevents atherosclerosis by skewing the immune response toward an antiinflammatory profile , 2010, The Journal of experimental medicine.

[21]  F. Geissmann,et al.  Monocytes in atherosclerosis: subsets and functions , 2010, Nature Reviews Cardiology.

[22]  Carl Nathan,et al.  Nonresolving Inflammation , 2010, Cell.

[23]  A. Heiligenhaus,et al.  Amniotic membrane transplantation induces apoptosis in T lymphocytes in murine corneas with experimental herpetic stromal keratitis. , 2009, Investigative ophthalmology & visual science.

[24]  Wei Li,et al.  Biochemical Characterization and Function of Complexes Formed by Hyaluronan and the Heavy Chains of Inter-α-inhibitor (HC·HA) Purified from Extracts of Human Amniotic Membrane* , 2009, The Journal of Biological Chemistry.

[25]  A. Sica,et al.  M2 Macrophages Phagocytose Rituximab-Opsonized Leukemic Targets More Efficiently than M1 Cells In Vitro1 , 2009, The Journal of Immunology.

[26]  I. Megson,et al.  Macrophage phagocytosis of apoptotic neutrophils is critically regulated by the opposing actions of pro‐inflammatory and anti‐inflammatory agents: key role for TNF‐α , 2009, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  J. Edwards,et al.  Exploring the full spectrum of macrophage activation , 2008, Nature Reviews Immunology.

[28]  Wei Li,et al.  Suppression of activation and induction of apoptosis in RAW264.7 cells by amniotic membrane extract. , 2008, Investigative ophthalmology & visual science.

[29]  Y. Mohri,et al.  Effects of a hyaluronate-carboxymethylcellulose membrane (Seprafilm) on human polymorphonuclear neutrophil functions. , 2008, The Journal of surgical research.

[30]  Carey L. Medin,et al.  Functional Characterization of Murine Interferon Regulatory Factor 5 (IRF-5) and Its Role in the Innate Antiviral Response* , 2008, Journal of Biological Chemistry.

[31]  S. Iijima,et al.  Siglec-9 enhances IL-10 production in macrophages via tyrosine-based motifs. , 2008, Biochemical and biophysical research communications.

[32]  P. Kubes,et al.  Interaction of CD44 and hyaluronan is the dominant mechanism for neutrophil sequestration in inflamed liver sinusoids , 2008, The Journal of experimental medicine.

[33]  A. Mantovani,et al.  PTX3 interacts with inter-alpha-trypsin inhibitor: implications for hyaluronan organization and cumulus oophorus expansion. , 2007, The Journal of biological chemistry.

[34]  A. Heiligenhaus,et al.  On the influence of neutrophils in corneas with necrotizing HSV-1 keratitis following amniotic membrane transplantation. , 2007, Experimental eye research.

[35]  P. Henson,et al.  Immunological consequences of apoptotic cell phagocytosis. , 2007, The American journal of pathology.

[36]  Frank Brombacher,et al.  Macrophage-specific PPARγ controls alternative activation and improves insulin resistance , 2007, Nature.

[37]  T. Oberyszyn.  Inflammation and wound healing. , 2007, Frontiers in bioscience : a journal and virtual library.

[38]  E. Vizi,et al.  Shaping of monocyte and macrophage function by adenosine receptors. , 2007, Pharmacology & therapeutics.

[39]  A. Saltiel,et al.  Obesity induces a phenotypic switch in adipose tissue macrophage polarization. , 2007, The Journal of clinical investigation.

[40]  J. Wallace,et al.  Resolution of in flammation: state of the art, definitions and terms , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[41]  K. Mchugh,et al.  Cloning and characterization of osteoclast precursors from the raw264.7 cell line , 2006, In Vitro Cellular & Developmental Biology - Animal.

[42]  J. Pugin.  The Role of the Macrophage , 2007 .

[43]  Xia Zhang,et al.  Biochemical and functional characterization of three activated macrophage populations , 2006, Journal of leukocyte biology.

[44]  E. Lasunskaia,et al.  Mycobacteria directly induce cytoskeletal rearrangements for macrophage spreading and polarization through TLR2‐dependent PI3K signaling , 2006, Journal of leukocyte biology.

[45]  John Savill,et al.  Requirements for Apoptotic Cell Contact in Regulation of Macrophage Responses1 , 2006, The Journal of Immunology.

[46]  Kate Schroder,et al.  Signal integration between IFNgamma and TLR signalling pathways in macrophages. , 2006, Immunobiology.

[47]  Christopher Haslett,et al.  Cyclin-dependent kinase inhibitors enhance the resolution of inflammation by promoting inflammatory cell apoptosis , 2006, Nature Medicine.

[48]  R. Kannagi,et al.  SHAP Potentiates the CD44-mediated Leukocyte Adhesion to the Hyaluronan Substratum* , 2006, Journal of Biological Chemistry.

[49]  Wei Li,et al.  Amniotic membrane induces apoptosis of interferon-γ activated macrophages in vitro , 2006 .

[50]  Wei Li,et al.  Amniotic membrane induces apoptosis of interferon-gamma activated macrophages in vitro. , 2006, Experimental eye research.

[51]  S. Gordon,et al.  Monocyte and macrophage heterogeneity , 2005, Nature Reviews Immunology.

[52]  L. Zitvogel,et al.  In vivo veritas , 2005, Nature Biotechnology.

[53]  A. Zwinderman,et al.  Synovial tissue macrophages: a sensitive biomarker for response to treatment in patients with rheumatoid arthritis , 2004, Annals of the rheumatic diseases.

[54]  J. Hiscott,et al.  A CRM1-dependent Nuclear Export Pathway Is Involved in the Regulation of IRF-5 Subcellular Localization* , 2005, Journal of Biological Chemistry.

[55]  T. Yamaguchi,et al.  Effects of hyaluronic acid on macrophage phagocytosis and active oxygen release , 2005, Agents and Actions.

[56]  Wei Li,et al.  How does amniotic membrane work? , 2004, The ocular surface.

[57]  C. Bouchard,et al.  Amniotic membrane transplantation in the management of severe ocular surface disease: indications and outcomes. , 2004, The ocular surface.

[58]  Marilyn S. Rugg,et al.  TSG-6 Modulates the Interaction between Hyaluronan and Cell Surface CD44*[boxs] , 2004, Journal of Biological Chemistry.

[59]  Adriano G. Rossi,et al.  Inflammatory Resolution: new opportunities for drug discovery , 2004, Nature Reviews Drug Discovery.

[60]  C. Garlanda,et al.  PTX3 plays a key role in the organization of the cumulus oophorus extracellular matrix and in in vivo fertilization , 2004, Development.

[61]  H. Dua,et al.  The amniotic membrane in ophthalmology. , 2004, Survey of ophthalmology.

[62]  M. Mörgelin,et al.  Molecular Heterogeneity of the SHAP-Hyaluronan Complex , 2003, Journal of Biological Chemistry.

[63]  S. Akira,et al.  Role of Adaptor TRIF in the MyD88-Independent Toll-Like Receptor Signaling Pathway , 2003, Science.

[64]  D. Mosser,et al.  The many faces of macrophage activation , 2003, Journal of leukocyte biology.

[65]  J. Duffield.  The inflammatory macrophage , 2003 .

[66]  S. Gordon.  Alternative activation of macrophages , 2003, Nature Reviews Immunology.

[67]  J. Duffield.  The inflammatory macrophage: a story of Jekyll and Hyde. , 2003, Clinical science.

[68]  P. Pitha,et al.  Multiple Regulatory Domains of IRF-5 Control Activation, Cellular Localization, and Induction of Chemokines That Mediate Recruitment of T Lymphocytes , 2002, Molecular and Cellular Biology.

[69]  D. Mosser,et al.  A novel phenotype for an activated macrophage: the type 2 activated macrophage , 2002, Journal of leukocyte biology.

[70]  L. Rizzo,et al.  Macrophages from IL‐12p40‐deficient mice have a bias toward the M2 activation profile , 2002, Journal of leukocyte biology.

[71]  David A. Hume,et al.  Generation of Diversity in the Innate Immune System: Macrophage Heterogeneity Arises from Gene-Autonomous Transcriptional Probability of Individual Inducible Genes1 , 2002, The Journal of Immunology.

[72]  T. Ahrens,et al.  The role of CD44 during CD40 ligand‐induced dendritic cell clustering and maturation , 2001, Journal of leukocyte biology.

[73]  A. Heiligenhaus,et al.  Improvement of HSV-1 necrotizing keratitis with amniotic membrane transplantation. , 2001, Investigative ophthalmology & visual science.

[74]  J. Gerber,et al.  Reversing Lipopolysaccharide Toxicity by Ligating the Macrophage Fcγ Receptors1 , 2001, The Journal of Immunology.

[75]  S. Shimmura,et al.  Antiinflammatory Effects of Amniotic Membrane Transplantation in Ocular Surface Disorders , 2001, Cornea.

[76]  W. Culbertson,et al.  Reduction in corneal haze and apoptosis by amniotic membrane matrix in excimer laser photoablation in rabbits , 2001, Journal of cataract and refractive surgery.

[77]  S. Frisch,et al.  Anoikis mechanisms. , 2001, Current opinion in cell biology.

[78]  S. Rhee,et al.  Murine TOLL-like Receptor 4 Confers Lipopolysaccharide Responsiveness as Determined by Activation of NFκB and Expression of the Inducible Cyclooxygenase* , 2000, Journal of Biological Chemistry.

[79]  S. Tseng,et al.  Modulation of acute inflammation and keratocyte death by suturing, blood, and amniotic membrane in PRK. , 2000, Investigative ophthalmology & visual science.

[80]  J. Lord,et al.  Spontaneous Neutrophil Apoptosis Involves Caspase 3-mediated Activation of Protein Kinase C-δ* , 1999, The Journal of Biological Chemistry.

[81]  A. Khwaja,et al.  Caspase-mediated proteolysis and activation of protein kinase Cdelta plays a central role in neutrophil apoptosis. , 1999, Blood.

[82]  G. Bokoch,et al.  Characterization of Rac and Cdc42 Activation in Chemoattractant-stimulated Human Neutrophils Using a Novel Assay for Active GTPases* , 1999, The Journal of Biological Chemistry.

[83]  E. Muñoz-Elías,et al.  Vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide prevent inducible nitric oxide synthase transcription in macrophages by inhibiting NF-kappa B and IFN regulatory factor 1 activation. , 1999, Journal of immunology.

[84]  A. Aderem,et al.  Mechanisms of phagocytosis in macrophages. , 1999, Annual review of immunology.

[85]  J. Nick,et al.  p38 Mitogen-activated Protein Kinase-dependent and -independent Intracellular Signal Transduction Pathways Leading to Apoptosis in Human Neutrophils* , 1998, The Journal of Biological Chemistry.

[86]  V. Fadok,et al.  Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF. , 1998, The Journal of clinical investigation.

[87]  R. Stern,et al.  A microtiter-based assay for hyaluronidase activity not requiring specialized reagents. , 1997, Analytical biochemistry.

[88]  S. Kurono,et al.  Evidence for the Covalent Binding of SHAP, Heavy Chains of Inter-α-Trypsin Inhibitor, to Hyaluronan (*) , 1995, The Journal of Biological Chemistry.

[89]  F. Finkelman,et al.  Evidence for the involvement of interleukin 10 in the differential deactivation of murine peritoneal macrophages by prostaglandin E2 , 1994, The Journal of experimental medicine.

[90]  H. Nochi,et al.  High‐Molecular‐Weight Hyaluronic Acids Inhibit Chemotaxis and Phagocytosis but Not Lysosomal Enzyme Release Induced by Receptor‐Mediated Stimulations in Guinea Pig Phagocytes , 1994, Microbiology and immunology.

[91]  C. Haslett,et al.  Inhibition of apoptosis and prolongation of neutrophil functional longevity by inflammatory mediators , 1993, Journal of leukocyte biology.

[92]  J. Vilček,et al.  TSG-14, a tumor necrosis factor- and IL-1-inducible protein, is a novel member of the pentaxin family of acute phase proteins. , 1993, Journal of immunology.

[93]  D. Mosser,et al.  Treatment of murine macrophages with interferon‐γ inhibits their ability to bind leishmania promastigotes , 1992, Journal of leukocyte biology.

[94]  S Gordon,et al.  Interleukin 4 potently enhances murine macrophage mannose receptor activity: a marker of alternative immunologic macrophage activation , 1992, The Journal of experimental medicine.

[95]  J. Enghild,et al.  Chondroitin 4-sulfate covalently cross-links the chains of the human blood protein pre-alpha-inhibitor. , 1991, The Journal of biological chemistry.

[96]  R. Schulte,et al.  Binding of hyaluronic acid to lymphoid cell lines is inhibited by monoclonal antibodies against Pgp-1. , 1990, Experimental cell research.

[97]  C. Haslett,et al.  Resolution of Inflammation , 1988 .

[98]  J. Forrester,et al.  Inhibition of phagocytosis by high molecular weight hyaluronate. , 1980, Immunology.