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Updated: Aug 9, 2026
Determination of Vaccine Immunogenicity Using Bovine Monocyte-Derived Dendritic Cells
Published on: May 19, 2023
Monocyte-derived dendritic cells from horses differ from dendritic cells of humans and mice
Susanne Mauel1, Falko Steinbach, Hanns Ludwig
1Institute of Virology, FU Berlin, Berlin, Germany.
Insights
Researchers cloned equine granulocyte-macrophage colony-stimulating factor (GM-CSF) and developed equine myeloid dendritic cells (MoDC). Equine MoDC showed typical morphology and function, but required longer differentiation times than human cells.
Area of Science:
- Immunology
- Cell Biology
- Equine Science
Background:
- Dendritic cells (DC) initiate immune responses and are found in most tissues.
- Myeloid dendritic cells (MoDC) are generated in vitro from monocytes using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4).
- Understanding the equine immune system is crucial for veterinary medicine.
Purpose of the Study:
- To clone and characterize equine (eq.) GM-CSF.
- To generate and characterize equine myeloid dendritic cells (eq.MoDC) in vitro.
- To compare the differentiation and function of eq.MoDC with those of other species.
Main Methods:
- Cloning of eq.GM-CSF cDNA using degenerated primers and RACE RT-PCR.
- Differentiation of equine monocytes into MoDC using eq.GM-CSF and eq.IL-4.
- Analysis of eq.MoDC morphology, phenotype, and function using light and electron microscopy, flow cytometry, and mixed lymphocyte reaction.
Main Results:
- A novel 24-nucleotide extended open reading frame of eq.GM-CSF was identified.
- Generated eq.MoDC exhibited typical dendritic cell morphology and function, including allogeneic T cell stimulation.
- Equine MoDC differentiation required 6-7 days, longer than in humans.
- Lipopolysaccharide or poly(I:C) alone were insufficient for full maturation of eq.MoDC.
Conclusions:
- The study successfully cloned eq.GM-CSF and generated functional eq.MoDC.
- Equine MoDC differentiation is distinct from the human system, requiring longer culture times.
- The equine immune system exhibits unique characteristics compared to murine or human systems.
Abstract:
Dendritic cells (DC) are the initiators of immune responses and are present in most tissues in vivo. To generate myeloid DC from monocytes (MoDC) in vitro the necessary cytokines are granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Using degenerated primers delineated from other species and rapid amplification of cDNA ends reverse transcription-polymerase chain reaction (RACE RT-PCR), the cDNA of equine (eq.) GM-CSF was cloned and found to have a point deletion at the 3'-end of eq.GM-CSF, resulting in a 24-nucleotide extended open reading frame not described in any species thus far. For differentiating eq.MoDC, monocytes were stimulated with eq.GM-CSF and eq.IL-4. The eq.MoDC was analysed by both light and electron microscopy and by flow cytometry and mixed lymphocyte reaction. The eq.MoDC obtained had the typical morphology and function of DC, including the ability to stimulate allogeneic T cells in a mixed lymphocyte reaction. In contrast to the human system, however, monocytes had to be differentiated for 6-7 days before immature DC were obtained. Our data also indicate that lipopolysaccharide or poly(I:C) alone are not sufficient to confer the full phenotypic transition into mature DC. Thus our study contributes to understanding the heterogeneity of immunity and adds important information on the equine immune system, which is clearly distinct from those of mice or man.
