Constitutive Macropinocytosis Allows TAP-dependent Major Histocompatibility Complex Class I Presentation of Exogenous Soluble Antigen by Bone Marrow-Derived Dendritic Cells

Eur J Immunol. 1997 Jan;27(1):280-8. doi: 10.1002/eji.1830270141.

Abstract

Dendritic cells expanded from mouse bone marrow (BMDC) with granulocyte/macrophage-colony-stimulating factor have potent T cell-stimulatory properties both in vitro and in vivo. This has been well documented for major histocompatibility complex (MHC) class II-restricted responses, and more recently using peptide-loaded and protein-pulsed DC for CD8 responses following adoptive transfer in mice. An unresolved question concerns the capacity of BMDC to present exogenous antigen on MHC class I molecules, an unconventional mode of MHC class I loading for which there is now considerable evidence, particularly in macrophages. Here, we show that BMDC exhibit high levels of macropinocytosis driven by constitutive membrane ruffling activity. Up to one-third of actively ruffling and macropinocytosing BMDC transferred pinocytosed horseradish peroxidase into the cytosol following a 15-min pulse, suggesting that they might be capable of presenting exogenous soluble antigen on MHC class I molecules. We show that BMDC presented exogenous ovalbumin to a T cell hybridoma more effectively, more rapidly, and at lower exogenous antigen concentrations than BM macrophages on a cell-for-cell basis. Presentation was TAP dependent, brefeldin A sensitive, and blocked by inhibitors of proteasomal processing, demonstrating use of the classical MHC class I pathway. Although effective presentation of exogenous antigen by BMDC occurred in the absence of agents which stimulate macropinocytosis, treatment with phorbol myristate acetate (PMA) enhanced both pinocytosis and MHC class I presentation by BMDC. Finally, PMA-stimulated BMDC exposed to exogenous ovalbumin in vitro were able to prime an antigen-specific cytotoxic T lymphocyte response following adoptive transfer in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 2
  • ATP-Binding Cassette Transporters / physiology*
  • Animals
  • Antigen-Presenting Cells / immunology*
  • Antigens / chemistry
  • Antigens / immunology*
  • Bone Marrow / immunology*
  • Bone Marrow Cells
  • Dendritic Cells / immunology*
  • Histocompatibility Antigens Class I / immunology*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Pinocytosis
  • Solubility

Substances

  • ATP Binding Cassette Transporter, Subfamily B, Member 2
  • ATP-Binding Cassette Transporters
  • Antigens
  • Histocompatibility Antigens Class I
  • TAP1 protein, human
  • Tap1 protein, mouse