Cells with dendritic cell morphology and immunophenotype, binuclear morphology, and immunosuppressive function in dendritic cell cultures

Cell Immunol. 2011;272(1):1-10. doi: 10.1016/j.cellimm.2011.09.012. Epub 2011 Oct 8.

Abstract

Culturing of human peripheral blood CD14 positive monocytes is a method for generation of dendritic cells (DCs) for experimental purposes or for use in clinical grade vaccines. When culturing human DCs in this manner for clinical vaccine production, we noticed that 5-10% of cells within the bulk culture were binuclear or multiple nuclear, but had typical dendritic cell morphology and immunophenotype. We refer to the cells as binuclear cells in dendritic cell cultures (BNiDCs). By using single cell PCR analysis of mitochondrial DNA polymorphisms we demonstrated that approximately 20-25% of cells in DC culture undergo a fusion event. Flow sorted BNiDC express low HLA-DR and IL-12p70, but high levels of IL-10. In mixed lymphocyte reactions, purified BNiDC suppressed lymphocyte proliferation. Blockade of dendritic cell-specific transmembrane protein (DC-STAMP) decreased the number of binuclear cells in DC cultures. BNiDC represent a potentially tolerogenic population within DC preparations for clinical use.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Antibodies / pharmacology
  • Cell Culture Techniques / methods*
  • Cell Differentiation / immunology
  • Cell Fusion
  • Cell Nucleus
  • Dendritic Cells / cytology
  • Dendritic Cells / immunology*
  • Dendritic Cells / metabolism
  • Flow Cytometry
  • HLA-DR Antigens / biosynthesis
  • HLA-DR Antigens / immunology
  • Humans
  • Immune Tolerance
  • Immunity*
  • Immunohistochemistry
  • Immunophenotyping
  • Immunosuppression / methods*
  • Interleukin-10 / biosynthesis
  • Interleukin-10 / immunology
  • Interleukin-12 / biosynthesis
  • Interleukin-12 / immunology
  • Lipopolysaccharide Receptors / biosynthesis
  • Lipopolysaccharide Receptors / immunology
  • Lymphocyte Activation / immunology
  • Lymphocyte Culture Test, Mixed
  • Membrane Proteins / antagonists & inhibitors*
  • Membrane Proteins / immunology
  • Membrane Proteins / metabolism
  • Monocytes / cytology
  • Monocytes / immunology*
  • Monocytes / metabolism
  • Single-Cell Analysis

Substances

  • Adaptor Proteins, Signal Transducing
  • Antibodies
  • DCSTAMP protein, human
  • HLA-DR Antigens
  • Lipopolysaccharide Receptors
  • Membrane Proteins
  • Interleukin-10
  • Interleukin-12