title: Low molecular weight hyaluronan mediated CD44 dependent induction of IL-6 and chemokines in human dermal fibroblasts potentiates innate immune response.
abstract: Complex regulation of the wound healing process involves multiple interactions among stromal tissue cells, inflammatory cells, and the extracellular matrix. Low molecular weight hyaluronan (LMW HA) derived from the degradation of high molecular weight hyaluronan (HMW HA) is suggested to activate cells involved in wound healing through interaction with HA receptors. In particular, receptor CD44 is suggested to mediate cell response to HA of different MW, being the main cell surface HA receptor in stromal tissue and immune cells. However, the response of dermal fibroblasts, the key players in granulation tissue formation within the wound healing process, to LMW HA and their importance for the activation of immune cells is unclear. In this study we show that LMW HA (4.3kDa) induced pro-inflammatory cytokine IL-6 and chemokines IL-8, CXCL1, CXCL2, CXCL6 and CCL8 gene expression in normal human dermal fibroblasts (NHDF) that was further confirmed by increased levels of IL-6 and IL-8 in cell culture supernatants. Conversely, NHDF treated by HMW HA revealed a tendency to decrease the gene expression of these cytokine and chemokines when compared to untreated control. The blockage of CD44 expression by siRNA resulted in the attenuation of IL-6 and chemokines expression in LMW HA treated NHDF suggesting the involvement of CD44 in LMW HA mediated NHDF activation. The importance of pro-inflammatory mediators produced by LMW HA triggered NHDF was evaluated by significant activation of blood leukocytes exhibited as increased production of IL-6 and TNF-α. Conclusively, we demonstrated a pro-inflammatory response of dermal fibroblasts to LMW HA that was transferred to leukocytes indicating the significance of LMW HA in the inflammatory process development during the wound healing process.
authors.affiliation: Contipro Biotech spol. s.r.o., Dolni Dobrouc, Czech Republic; Institute of Experimental Biology, Department of Physiology and Immunology of Animals, Faculty of Science, Masaryk University, Brno, Czech Republic; Biomedical Center, Medical Faculty in Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Vistejnova
authors.affiliation: Contipro Biotech spol. s.r.o., Dolni Dobrouc, Czech Republic; Institute of Experimental Biology, Department of Physiology and Immunology of Animals, Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Barbora
authors.initials: B
authors.lastname: Safrankova
authors.affiliation: Contipro Biotech spol. s.r.o., Dolni Dobrouc, Czech Republic; Institute of Experimental Biology, Department of Physiology and Immunology of Animals, Faculty of Science, Masaryk University, Brno, Czech Republic.
authors.forename: Kristina
authors.initials: K
authors.lastname: Nesporova
authors.affiliation: Contipro Biotech spol. s.r.o., Dolni Dobrouc, Czech Republic.
authors.forename: Rastislav
authors.initials: R
authors.lastname: Slavkovsky
authors.affiliation: Contipro Biotech spol. s.r.o., Dolni Dobrouc, Czech Republic.
authors.forename: Martina
authors.initials: M
authors.lastname: Hermannova
authors.affiliation: Biomedical Center, Medical Faculty in Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Hosek
authors.affiliation: Contipro Biotech spol. s.r.o., Dolni Dobrouc, Czech Republic.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Velebny
authors.affiliation: Institute of Biophysics, Academy of Science of the Czech Republic v.v.i., Czech Republic; International Clinical Research Center - Center of Biomolecular and Cellular Engineering, St. Anne's University Hospital Brno, Brno, Czech Republic. Electronic address: kubalal@ibp.cz.
authors.forename: Lukas
authors.initials: L
authors.lastname: Kubala
mesh_terms: D018925:Chemokines / Q000096:biosynthesis / Q000235:genetics; D020405:Dermis / Q000166:cytology*; D005347:Fibroblasts / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000494:pharmacology*; D007113:Immunity, Innate / Q000187:drug effects* / Q000235:genetics; D018836:Inflammation Mediators / Q000378:metabolism; D015850:Interleukin-6 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D016209:Interleukin-8 / Q000096:biosynthesis; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism; D008970:Molecular Weight; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D018925:Chemokines; D018960:Hyaluronan Receptors; D018836:Inflammation Mediators; D015850:Interleukin-6; D016209:Interleukin-8; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; D006820:Hyaluronic Acid
keywords: Chemokines; Hyaluronan; IL-6; IL-8; Inflammation