All Title Author
Keywords Abstract

PLOS ONE  2011 

High and Low Molecular Weight Hyaluronic Acid Differentially Regulate Human Fibrocyte Differentiation

DOI: 10.1371/journal.pone.0026078

Full-Text   Cite this paper   Add to My Lib


Background Following tissue injury, monocytes can enter the tissue and differentiate into fibroblast-like cells called fibrocytes, but little is known about what regulates this differentiation. Extracellular matrix contains high molecular weight hyaluronic acid (HMWHA; ~2×106 Da). During injury, HMWHA breaks down to low molecular weight hyaluronic acid (LMWHA; ~0.8–8×105 Da). Methods and Findings In this report, we show that HMWHA potentiates the differentiation of human monocytes into fibrocytes, while LMWHA inhibits fibrocyte differentiation. Digestion of HMWHA with hyaluronidase produces small hyaluronic acid fragments, and these fragments inhibit fibrocyte differentiation. Monocytes internalize HMWHA and LMWHA equally well, suggesting that the opposing effects on fibrocyte differentiation are not due to differential internalization of HMWHA or LMWHA. Adding HMWHA to PBMC does not appear to affect the levels of the hyaluronic acid receptor CD44, whereas adding LMWHA decreases CD44 levels. The addition of anti-CD44 antibodies potentiates fibrocyte differentiation, suggesting that CD44 mediates at least some of the effect of hyaluronic acid on fibrocyte differentiation. The fibrocyte differentiation-inhibiting factor serum amyloid P (SAP) inhibits HMWHA-induced fibrocyte differentiation and potentiates LMWHA-induced inhibition. Conversely, LMWHA inhibits the ability of HMWHA, interleukin-4 (IL-4), or interleukin-13 (IL-13) to promote fibrocyte differentiation. Conclusions We hypothesize that hyaluronic acid signals at least in part through CD44 to regulate fibrocyte differentiation, with a dominance hierarchy of SAP>LMWHA≥HMWHA>IL-4 or IL-13.


[1]  Martin P (1997) Wound healing–aiming for perfect skin regeneration. Science 276: 75–81.
[2]  Singer AJ, Clark RA (1999) Cutaneous wound healing. N Engl J Med 341: 738–746.
[3]  Bucala R, Spiegel LA, Chesney J, Hogan M, Cerami A (1994) Circulating fibrocytes define a new leukocyte subpopulation that mediates tissue repair. Mol Med 1: 71–81.
[4]  Abe R, Donnelly SC, Peng T, Bucala R, Metz CN (2001) Peripheral blood fibrocytes: differentiation pathway and migration to wound sites. J Immunol 166: 7556–7562.
[5]  Yang L, Scott PG, Giuffre J, Shankowsky HA, Ghahary A, et al. (2002) Peripheral blood fibrocytes from burn patients: identification and quantification of fibrocytes in adherent cells cultured from peripheral blood mononuclear cells. Lab Invest 82: 1183–1192.
[6]  Pilling D, Buckley CD, Salmon M, Gomer RH (2003) Inhibition of fibrocyte differentiation by serum amyloid P. J Immunol 171: 5537–5546.
[7]  Maharjan AS, Pilling D, Gomer RH (2010) Toll-like receptor 2 agonists inhibit human fibrocyte differentiation. Fibrogenesis Tissue Repair 3: 23.
[8]  Shao DD, Suresh R, Vakil V, Gomer RH, Pilling D (2008) Pivotal Advance: Th-1 cytokines inhibit, and Th-2 cytokines promote fibrocyte differentiation. J Leukoc Biol 83: 1323–1333.
[9]  Phillips RJ, Burdick MD, Hong K, Lutz MA, Murray LA, et al. (2004) Circulating fibrocytes traffic to the lungs in response to CXCL12 and mediate fibrosis. J Clin Invest 114: 438–446.
[10]  Strieter RM, Keeley EC, Burdick MD, Mehrad B (2009) The role of circulating mesenchymal progenitor cells, fibrocytes, in promoting pulmonary fibrosis. Trans Am Clin Climatol Assoc 120: 49–59.
[11]  Reilkoff RA, Bucala R, Herzog EL (2011) Fibrocytes: emerging effector cells in chronic inflammation. Nat Rev Immunol 11: 427–435.
[12]  Moore BB, Kolodsick JE, Thannickal VJ, Cooke K, Moore TA, et al. (2005) CCR2-mediated recruitment of fibrocytes to the alveolar space after fibrotic injury. Am J Pathol 166: 675–684.
[13]  Hashimoto N, Jin H, Liu T, Chensue SW, Phan SH (2004) Bone marrow-derived progenitor cells in pulmonary fibrosis. J Clin Invest 113: 243–252.
[14]  Chesney J, Bucala R (1997) Peripheral blood fibrocytes: novel fibroblast-like cells that present antigen and mediate tissue repair. Biochem Soc Trans 25: 520–524.
[15]  Pilling D, Vakil V, Gomer RH (2009) Improved serum-free culture conditions for the differentiation of human and murine fibrocytes. J Immunol Meth 351: 62–70.
[16]  Gomperts BN, Strieter RM (2007) Fibrocytes in lung disease. J Leukoc Biol 82: 449–456.
[17]  Quan TE, Cowper S, Wu SP, Bockenstedt LK, Bucala R (2004) Circulating fibrocytes: collagen-secreting cells of the peripheral blood. Int J Biochem Cell Biol 36: 598–606.
[18]  Mori L, Bellini A, Stacey MA, Schmidt M, Mattoli S (2005) Fibrocytes contribute to the myofibroblast population in wounded skin and originate from the bone marrow. Exp Cell Res 304: 81–90.
[19]  Wang JF, Jiao H, Stewart TL, Shankowsky HA, Scott PG, et al. (2007) Fibrocytes from burn patients regulate the activities of fibroblasts. Wound Repair Regen 15: 113–121.
[20]  Hartlapp I, Abe R, Saeed RW, Peng T, Voelter W, et al. (2001) Fibrocytes induce an angiogenic phenotype in cultured endothelial cells and promote angiogenesis in vivo. FASEB J 15: 2215–2224.
[21]  Mattoli S, Bellini A, Schmidt M (2009) The role of a human hematopoietic mesenchymal progenitor in wound healing and fibrotic diseases and implications for therapy. Curr Stem Cell Res Ther 4: 266–280.
[22]  Bellini A, Mattoli S (2007) The role of the fibrocyte, a bone marrow-derived mesenchymal progenitor, in reactive and reparative fibroses. Lab Invest 87: 858–870.
[23]  Herzog EL, Bucala R (2010) Fibrocytes in health and disease. Exp Hematol 38: 548–556.
[24]  Jiang D, Liang J, Noble PW (2007) Hyaluronan in Tissue Injury and Repair. Ann Rev Cell Dev Biol 23: 435–461.
[25]  McKee CM, Penno MB, Cowman M, Burdick MD, Strieter RM, et al. (1996) Hyaluronan (HA) fragments induce chemokine gene expression in alveolar macrophages. The role of HA size and CD44. J Clin Invest 98: 2403–2413.
[26]  Li Y, Jiang D, Liang J, Meltzer EB, Gray A, et al. (2011) Severe lung fibrosis requires an invasive fibroblast phenotype regulated by hyaluronan and CD44. J Exp Med. doi:10.1084/jem.20102510.
[27]  Laurent TC, Fraser JR (1992) Hyaluronan. FASEB J 7: 2397–2404.
[28]  Teder P, Vandivier RW, Jiang D, Liang J, Cohn L, et al. (2002) Resolution of lung inflammation by CD44. Science 296: 155–158.
[29]  Kuo J-W (2005) Practical Aspects of Hyaluronan Based Medical Products. Boston: Taylor and Francis. pp. 1–209.
[30]  Filion MC, Phillips NC (2001) Pro-inflammatory activity of contaminating DNA in hyaluronic acid preparations. J Pharm Pharmacol 53: 555–561.
[31]  Day AJ, de la Motte CA (2005) Hyaluronan cross-linking: a protective mechanism in inflammation? Trends Immunol 26: 637–643.
[32]  Termeer C, Benedix F, Sleeman J, Fieber C, Voith U, et al. (2002) Oligosaccharides of Hyaluronan activate dendritic cells via toll-like receptor 4. J Exp Med 195: 99–111.
[33]  Scheibner KA, Lutz MA, Boodoo S, Fenton MJ, Powell JD, et al. (2006) Hyaluronan fragments act as an endogenous danger signal by engaging TLR2. J Immunol 177: 1272–1281.
[34]  Bollyky PL, Lord JD, Masewicz SA, Evanko SP, Buckner JH, et al. (2007) Cutting edge: high molecular weight hyaluronan promotes the suppressive effects of CD4+CD25+ regulatory T cells. J Immunol 179: 744–747.
[35]  Chang EJ, Kim HJ, Ha J, Ryu J, Park KH, et al. (2007) Hyaluronan inhibits osteoclast differentiation via Toll-like receptor 4. J Cell Sci 120: 166–176.
[36]  Termeer CC, Hennies J, Voith U, Ahrens T, Weiss JM, et al. (2000) Oligosaccharides of hyaluronan are potent activators of dendritic cells. J Immunol 165: 1863–1870.
[37]  Nakamura K, Yokohama S, Yoneda M, Okamoto S, Tamaki Y, et al. (2004) High, but not low, molecular weight hyaluronan prevents T-cell-mediated liver injury by reducing proinflammatory cytokines in mice. J Gastroenterol 39: 346–354.
[38]  Jiang D, Liang J, Fan J, Yu S, Chen S, et al. (2005) Regulation of lung injury and repair by Toll-like receptors and hyaluronan. Nat Med 11: 1173–1179.
[39]  Peach RJ, Hollenbaugh D, Stamenkovic I, Aruffo A (1993) Identification of hyaluronic acid binding sites in the extracellular domain of CD44. J Cell Biol 122: 257–264.
[40]  Taylor KR, Yamasaki K, Radek KA, Di Nardo A, Goodarzi H, et al. (2007) Recognition of hyaluronan released in sterile injury involves a unique receptor complex dependent on Toll-like receptor 4, CD44, and MD-2. J Biol Chem 282: 18265–18275.
[41]  Siegelman MH, DeGrendele HC, Estess P (1999) Activation and interaction of CD44 and hyaluronan in immunological systems. J Leukoc Biol 66: 315–321.
[42]  Svee K, White J, Vaillant P, Jessurun J, Roongta U, et al. (1996) Acute lung injury fibroblast migration and invasion of a fibrin matrix is mediated by CD44. J Clin Invest 98: 1713–1727.
[43]  Campo GM, Avenoso A, Campo S, D'Ascola A, Nastasi G, et al. (2010) Molecular size hyaluronan differently modulates toll-like receptor-4 in LPS-induced inflammation in mouse chondrocytes. Biochimie 92: 204–215.
[44]  Zaman A, Cui Z, Foley JP, Zhao H, Grimm PC, et al. (2005) Expression and role of the hyaluronan receptor RHAMM in inflammation after bleomycin injury. Am J Resp Cell Mol Biol 33: 447–454.
[45]  Hamilton SR, Fard SF, Paiwand FF, Tolg C, Veiseh M, et al. (2007) The hyaluronan receptors CD44 and Rhamm (CD168) form complexes with ERK1,2 that sustain high basal motility in breast cancer cells. J Biol Chem 282: 16667–16680.
[46]  Johnson LA, Prevo R, Clasper S, Jackson DG (2007) Inflammation-induced uptake and degradation of the lymphatic endothelial hyaluronan receptor LYVE-1. J Biol Chem 282: 33671–33680.
[47]  Pilling D, Tucker NM, Gomer RH (2006) Aggregated IgG inhibits the differentiation of human fibrocytes. J Leukoc Biol 79: 1242–1251.
[48]  Trujillo G, Meneghin A, Flaherty KR, Sholl LM, Myers JL, et al. (2010) TLR9 differentiates rapidly from slowly progressing forms of idiopathic pulmonary fibrosis. Sci Transl Med 2: 57ra82.
[49]  Wang J, Jiao H, Stewart TL, Shankowsky HA, Scott PG, et al. (2007) Improvement in postburn hypertrophic scar after treatment with IFN-alpha2b is associated with decreased fibrocytes. J Interferon Cytokine Res 27: 921–930.
[50]  Kansas GS, Wood GS, Dailey MO (1989) A family of cell-surface glycoproteins defined by a putative anti-endothelial cell receptor antibody in man. J Immunol 142: 3050–3057.
[51]  Lee HG, Cowman MK (1994) An agarose gel electrophoretic method for analysis of hyaluronan molecular weight distribution. Anal Biochem 219: 278–287.
[52]  Sharp PA, Sugden B, Sambrook J (1973) Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose–ethidium bromide electrophoresis. Biochemistry 12: 3055–3063.
[53]  Pilling D FT, Huang D, Kaul B, Gomer RH (2009) Identification of Markers that Distinguish Monocyte-Derived Fibrocytes from Monocytes, Macrophages, and Fibroblasts. PLoS ONE 4: e7475.
[54]  Girish KS, Kemparaju K (2007) The magic glue hyaluronan and its eraser hyaluronidase: A biological overview. Life Sciences 80: 1921–1943.
[55]  Stern R, Asari AA, Sugahara KN (2006) Hyaluronan fragments: An information-rich system. Eur J Cell Biol 85: 699–715.
[56]  Powell JD, Horton MR (2005) Threat matrix: low-molecular-weight hyaluronan (HA) as a danger signal. Immunol Res 31: 207–218.
[57]  Lesley J, Hyman R, Kincade PW (1993) CD44 and its interaction with extracellular matrix. Adv Immunol 54: 271–335.
[58]  Nong YH, Remold-O'Donnell E, LeBien TW, Remold HG (1989) A monoclonal antibody to sialophorin (CD43) induces homotypic adhesion and activation of human monocytes. J Exp Med 170: 259–267.
[59]  Hardwick C, Hoare K, Owens R, Hohn HP, Hook M, et al. (1992) Molecular cloning of a novel hyaluronan receptor that mediates tumor cell motility. J Cell Biol 117: 1343–1350.
[60]  Nedvetzki S, Gonen E, Assayag N, Reich R, Williams RO, et al. (2004) RHAMM, a receptor for hyaluronan-mediated motility, compensates for CD44 in inflamed CD44-knockout mice: a different interpretation of redundancy. Proc Natl Acad Sci U S A 101: 18081–18086.
[61]  Weiss JM, Renkl AC, Ahrens T, Moll J, Mai BH, et al. (1998) Activation-dependent modulation of hyaluronate-receptor expression and of hyaluronate-avidity by human monocytes. J Invest Dermatol 111: 227–232.
[62]  Greiner J, Ringhoffer M, Taniguchi M, Schmitt A, Kirchner D, et al. (2002) Receptor for hyaluronan acid-mediated motility (RHAMM) is a new immunogenic leukemia-associated antigen in acute and chronic myeloid leukemia. Exp Hematol 30: 1029–1035.
[63]  Engstrom-Laurent A, Loof L, Nyberg A, Schroder T (1985) Increased serum levels of hyaluronate in liver disease. Hepatology 5: 638–642.
[64]  Bianchetti L, Barczyk M, Cardoso J, Schmidt M, Bellini A, et al. (2011) Extracellular matrix remodelling properties of human fibrocytes. J Cell Mol Med. doi:10.1111/j.1582-4934.
[65]  Niedermeier M, Reich B, Rodriguez Gomez M, Denzel A, Schmidbauer K, et al. (2009) CD4+ T cells control the differentiation of Gr1+ monocytes into fibrocytes. Proc Natl Acad Sci U S A 106: 17892–17897.
[66]  Naik-Mathuria B, Pilling D, Crawford JR, Gay AN, Smith CW, et al. (2008) Serum amyloid P inhibits dermal wound healing. Wound Repair Regen 16: 266–273.
[67]  Pilling D, Roife D, Wang M, Ronkainen S, Crawford JR, et al. (2007) Reduction of Bleomycin-Induced Pulmonary Fibrosis by Serum Amyloid P. J Immunol 179: 4035–4044.
[68]  Murray LA, Chen Q, Kramer MS, Hesson DP, Argentieri RL, et al. (2011) TGF-beta driven lung fibrosis is macrophage dependent and blocked by Serum amyloid P. Int J Biochem Cell Biol 43: 154–162.
[69]  Murray LA, Rosada R, Moreira AP, Joshi A, Kramer MS, et al. (2010) Serum amyloid P therapeutically attenuates murine bleomycin-induced pulmonary fibrosis via its effects on macrophages. PLoS One 5: e9683.
[70]  Grishko V, Xu M, Ho R, Mates A, Watson S, et al. (2009) Effects of hyaluronic acid on mitochondrial function and mitochondria-driven apoptosis following oxidative stress in human chondrocytes. J Biol Chem 284: 9132–9139.
[71]  Forrester JV, Balazs EA (1980) Inhibition of phagocytosis by high molecular weight hyaluronate. Immunology 40: 435–446.
[72]  Kuang DM, Wu Y, Chen N, Cheng J, Zhuang SM, et al. (2007) Tumor-derived hyaluronan induces formation of immunosuppressive macrophages through transient early activation of monocytes. Blood 110: 587–595.
[73]  Hodge-Dufour J, Noble PW, Horton MR, Bao C, Wysoka M, et al. (1997) Induction of IL-12 and chemokines by hyaluronan requires adhesion-dependent priming of resident but not elicited macrophages. J Immunol 159: 2492–2500.
[74]  Bai KJ, Spicer AP, Mascarenhas MM, Yu L, Ochoa CD, et al. (2005) The role of hyaluronan synthase 3 in ventilator-induced lung injury. Am J Respir Crit Care Med 172: 92–98.


comments powered by Disqus