title: Endotoxin free hyaluronan and hyaluronan fragments do not stimulate TNF-α, interleukin-12 or upregulate co-stimulatory molecules in dendritic cells or macrophages.
abstract: The extracellular matrix glycosaminoglycan, hyaluronan, has been described as a regulator of tissue inflammation, with hyaluronan fragments reported to stimulate innate immune cells. High molecular mass hyaluronan is normally present in tissues, but upon inflammation lower molecular mass fragments are generated. It is unclear if these hyaluronan fragments induce an inflammatory response or are a consequence of inflammation. In this study, mouse bone marrow derived macrophages and dendritic cells (DCs) were stimulated with various sizes of hyaluronan from different sources, fragmented hyaluronan, hyaluronidases and heavy chain modified-hyaluronan (HA-HC). Key pro-inflammatory molecules, tumour necrosis factor alpha, interleukin-1 beta, interleukin-12, CCL3, and the co-stimulatory molecules, CD40 and CD86 were measured. Only human umbilical cord hyaluronan, bovine testes and Streptomyces hyaluronlyticus hyaluronidase stimulated macrophages and DCs, however, these reagents were found to be contaminated with endotoxin, which was not fully removed by polymyxin B treatment. In contrast, pharmaceutical grade hyaluronan and hyaluronan fragments failed to stimulate in vitro-derived or ex vivo macrophages and DCs, and did not induce leukocyte recruitment after intratracheal instillation into mouse lungs. Hence, endotoxin-free pharmaceutical grade hyaluronan does not stimulate macrophages and DCs in our inflammatory models. These results emphasize the importance of ensuring hyaluronan preparations are endotoxin free.
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada.
authors.forename: Arif
authors.initials: A
authors.lastname: Arif
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada.|Department of Pediatrics, British Columbia Children's Hospital Research Institute, Vancouver, B.C. Canada.
authors.forename: Mia
authors.initials: M
authors.lastname: Olsson
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio 44195, U.S.A.
authors.forename: Valbona
authors.initials: V
authors.lastname: Cali
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada.
authors.forename: Blair
authors.initials: B
authors.lastname: Hardman
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada.
authors.forename: Manisha
authors.initials: M
authors.lastname: Dosanjh
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio 44195, U.S.A.
authors.forename: Mark
authors.initials: M
authors.lastname: Lauer
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio 44195, U.S.A.
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Midura
authors.affiliation: Department of Biomedical Engineering, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio 44195, U.S.A.
authors.forename: Vincent C
authors.initials: VC
authors.lastname: Hascall
authors.affiliation: Department of Pediatrics, British Columbia Children's Hospital Research Institute, Vancouver, B.C. Canada.
authors.forename: Kelly L
authors.initials: KL
authors.lastname: Brown
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, B.C. V6T 1Z3, Canada.
authors.forename: Pauline
authors.initials: P
authors.lastname: Johnson
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology*; D006820:Hyaluronic Acid / Q000378:metabolism*; D018664:Interleukin-12 / Q000096:biosynthesis*; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology*; D051379:Mice; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
chemical_list: D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12; D006820:Hyaluronic Acid