Hyaluronan Binding Identifies a Functionally Distinct Alveolar Macrophage-like Population in Bone Marrow-Derived Dendritic Cell Cultures

J Immunol. 2015 Jul 15;195(2):632-42. doi: 10.4049/jimmunol.1402506. Epub 2015 Jun 17.


Although classical dendritic cells (DCs) arise from distinct progenitors in the bone marrow, the origin of inflammatory DCs and the distinction between monocyte-derived DCs and macrophages is less clear. In vitro culture of mouse bone marrow cells with GM-CSF is a well-established method to generate DCs, but GM-CSF has also been used to generate bone marrow-derived macrophages. In this article, we identify a distinct subpopulation of cells within the GM-CSF bone marrow-derived DC culture based on their ability to bind hyaluronan (HA), a major component of the extracellular matrix and ligand for CD44. HA identified a morphologically distinct subpopulation of cells within the immature DC population (CD11c(+) MHC II(mid/low)) that were CCR5(+)/CCR7(-) and proliferated in response to GM-CSF, but, unlike immature DCs, did not develop into mature DCs expressing CCR7 and high levels of MHC II, even after stimulation with LPS. The majority of these cells produced TNF-α in response to LPS but were unable to activate naive T cells, whereas the majority of mature DCs produced IL-12 and activated naive T cells. This HA binding population shared many characteristics with alveolar macrophages and was retained in the alveolar space after lung instillation even after LPS stimulation, whereas the MHC II(high) mature DCs were found in the draining lymph node. Thus, HA binding in combination with MHC II expression can be used to identify alveolar-like macrophages from GM-CSF-treated bone marrow cultures, which provides a useful in vitro model to study alveolar macrophages.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Marrow Cells / cytology*
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / immunology
  • CD11c Antigen / genetics
  • CD11c Antigen / immunology
  • Cell Differentiation / drug effects
  • Cell Lineage / drug effects
  • Cell Lineage / immunology*
  • Cell Proliferation / drug effects
  • Dendritic Cells / cytology*
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology
  • Gene Expression
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Hyaluronan Receptors / genetics
  • Hyaluronan Receptors / immunology
  • Hyaluronic Acid / metabolism*
  • Hyaluronic Acid / pharmacology
  • Lipopolysaccharides / pharmacology
  • Lung / cytology
  • Lung / drug effects
  • Lung / immunology
  • Lymph Nodes / cytology
  • Lymph Nodes / drug effects
  • Lymph Nodes / immunology
  • Lymphocyte Activation / drug effects
  • Macrophages, Alveolar / cytology*
  • Macrophages, Alveolar / drug effects
  • Macrophages, Alveolar / immunology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Primary Cell Culture
  • Receptors, CCR5 / genetics
  • Receptors, CCR5 / immunology
  • Receptors, CCR7 / deficiency
  • Receptors, CCR7 / genetics
  • Receptors, CCR7 / immunology
  • T-Lymphocytes / cytology
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / immunology
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / immunology


  • CCR5 protein, mouse
  • CD11c Antigen
  • Ccr7 protein, mouse
  • Cd44 protein, mouse
  • Hyaluronan Receptors
  • Lipopolysaccharides
  • Receptors, CCR5
  • Receptors, CCR7
  • Tumor Necrosis Factor-alpha
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Hyaluronic Acid