title: Immobilized heavy chain-hyaluronic acid polarizes lipopolysaccharide-activated macrophages toward M2 phenotype.
abstract: 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.
authors.affiliation: From the TissueTech, Inc.
authors.forename: Hua
authors.initials: H
authors.lastname: He
authors.affiliation: From the TissueTech, Inc.
authors.forename: Suzhen
authors.initials: S
authors.lastname: Zhang
authors.affiliation: From the TissueTech, Inc.
authors.forename: Sean
authors.initials: S
authors.lastname: Tighe
authors.affiliation: the Ocular Surface Research Education Foundation, and.
authors.forename: Ji
authors.initials: J
authors.lastname: Son
authors.affiliation: From the TissueTech, Inc.,; the Ocular Surface Research Education Foundation, and; the Ocular Surface Center, Miami, Florida 33173. Electronic address: stseng@ocularsurface.com.
authors.forename: Scheffer C G
authors.initials: SCG
authors.lastname: Tseng
mesh_terms: D000276:Adjuvants, Immunologic / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis; D015536:Down-Regulation / Q000187:drug effects; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism* / Q000494:pharmacology; D050835:Interferon Regulatory Factors / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation / Q000187:drug effects*; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis; D047495:PPAR gamma / Q000096:biosynthesis; D010587:Phagocytosis / Q000187:drug effects; D051193:Toll-Like Receptors / Q000819:agonists / Q000235:genetics / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; D016207:Cytokines; C431713:IRF5 protein, human; D050835:Interferon Regulatory Factors; C498178:Irf5 protein, mouse; D008070:Lipopolysaccharides; D047495:PPAR gamma; D051193:Toll-Like Receptors; D006820:Hyaluronic Acid; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse
keywords: Amniotic Membrane, HC-HA, M1/M2 Phenotype; Apoptosis; Cell Adhesion; Cell Proliferation; Inflammation; Macrophages; Phagocytosis