Cytokine production and T-cell activation by macrophage-dendritic cells generated for therapeutic use

Br J Haematol. 2001 Sep;114(3):671-80. doi: 10.1046/j.1365-2141.2001.02982.x.


Clinical grade ex vivo-generated antigen-presenting cells, macrophage-dendritic cells (MAC-DCs) or macrophage-activated killers (MAKs) were derived from peripheral blood mononuclear cells (PBMCs). Cultures (7 d) were performed in non-adherent conditions in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and either interleukin 13 (IL-13) or dihydroxy-vitamin D3 respectively. MAKs were activated during the last 24 h with interferon gamma (IFNgamma). Reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) analyses indicated that IL-1beta and tumour necrosis factor alpha (TNFalpha) were produced by both cells. Higher pro-inflammatory cytokine (IL-1beta and TNFalpha) amounts were detected on average in MAK supernatants. In contrast, IL-12 p40 was found only in MAC-DC supernatants, but the biologically active IL-12 form (p70) was never detected. T-cell cytokines (IL-2, IL-4, IL-10) were not produced in culture conditions in which T cells were nevertheless present. At d 7, TNFalpha or lipopolysaccharide (LPS) upregulated IL-12 p40 production by MAC-DCs, while IL-12 p70 remained undetectable. LPS stimulation also increased TNFalpha production by these cells. Allogeneic mixed lymphocyte reactions (MLR) showed that MAKs are poor stimulatory cells compared with MAC-DCs. The MAC-DC stimulatory capacity was enhanced by LPS, although the expression of HLA class II, CD83, CD80 and CD86 was unmodified. Thus, MAC-DCs represent a tool for triggering adaptative immunity, while MAK should be primarily used as effector killer cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 24,25-Dihydroxyvitamin D 3 / pharmacology
  • Cells, Cultured
  • Cytokines / biosynthesis*
  • Dendritic Cells / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Fluorescent Antibody Technique, Direct
  • Fluorescent Antibody Technique, Indirect
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Humans
  • Immunotherapy, Adoptive
  • Interleukin-1 / biosynthesis
  • Interleukin-1 / genetics
  • Interleukin-12 / biosynthesis
  • Interleukin-12 / genetics
  • Interleukin-13 / pharmacology
  • Lipopolysaccharides / pharmacology
  • Lymphocyte Activation*
  • Macrophage Activation
  • Macrophages / immunology*
  • RNA, Messenger / analysis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stimulation, Chemical
  • T-Lymphocytes / immunology*
  • Tumor Necrosis Factor-alpha / biosynthesis
  • Tumor Necrosis Factor-alpha / genetics


  • Cytokines
  • Interleukin-1
  • Interleukin-13
  • Lipopolysaccharides
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha
  • Interleukin-12
  • 24,25-Dihydroxyvitamin D 3
  • Granulocyte-Macrophage Colony-Stimulating Factor