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The effect of CpG-ODN on antigen presenting cells of the foal
  • Cornell University
  • Universidade Estadual Paulista (UNESP)
  • University of Kentucky
  • Qiagen GmbH
Background: Cytosine-phosphate-guanosine oligodeoxynucleotide (CpG-ODN) has been used successfully to induce immune responses against viral and intracellular organisms in mammals. The main objective of this study was to test the effect of CpG-ODN on antigen presenting cells of young foals. Methods: Peripheral blood monocytes of foals (n = 7) were isolated in the first day of life and monthly thereafter up to 3 months of life. Adult horse (n = 7) monocytes were isolated and tested once for comparison. Isolated monocytes were stimulated with IL-4 and GM-CSF (to obtain dendritic cells, DC) or not stimulated (to obtain macrophages). Macrophages and DCs were stimulated for 14-16 hours with either CpG-ODN, LPS or not stimulated. The stimulated and non-stimulated cells were tested for cell surface markers (CD86 and MHC class II) using flow cytometry, mRNA expression of cytokines (IL-12, IFNα, IL-10) and TLR-9 using real time quantitative RT-PCR, and for the activation of the transcription factor NF-κB p65 using a chemiluminescence assay. Results: The median fluorescence of the MHC class II molecule in non-stimulated foal macrophages and DCs at birth were 12.5 times and 11.2 times inferior, respectively, than adult horse cells (p = 0.009). That difference subsided at 3 months of life (p = 0.3). The expression of the CD86 co-stimulatory molecule was comparable in adult horse and foal macrophages and DCs, independent of treatment. CpG-ODN stimulation induced IL-12p40 (53 times) and IFNα (23 times) mRNA expression in CpG-ODN-treated adult horse DCs (p = 0.078), but not macrophages, in comparison to non-stimulated cells. In contrast, foal APCs did not respond to CpG-ODN stimulation with increased cytokine mRNA expression up to 3 months of age. TLR-9 mRNA expression and NF-kB activation (NF-kB p65) in foal DCs and macrophages were comparable (p > 0.05) to adult horse cells. Conclusion: CpG-ODN treatment did not induce specific maturation and cytokine expression in foal macrophages and DCs. Nevertheless, adult horse DCs, but not macrophages, increased their expression of IL-12 and IFNα cytokines upon CpG-ODN stimulation. Importantly, foals presented an age-dependent limitation in the expression of MHC class II in macrophages and DCs, independent of treatment. © 2007 Flaminio et al; licensee BioMed Central Ltd.
Issue Date: 
Journal of Immune Based Therapies and Vaccines, v. 5.
  • alpha interferon
  • CD86 antigen
  • cell surface marker
  • CpG oligodeoxynucleotide
  • cytokine
  • granulocyte macrophage colony stimulating factor
  • immunoglobulin enhancer binding protein
  • interleukin 10
  • interleukin 12
  • interleukin 4
  • lipopolysaccharide
  • major histocompatibility antigen class 2
  • messenger RNA
  • toll like receptor 9
  • animal cell
  • antigen presenting cell
  • cell isolation
  • cell stimulation
  • chemoluminescence
  • controlled study
  • dendritic cell
  • flow cytometry
  • foal
  • gene expression
  • horse
  • macrophage
  • monocyte
  • nonhuman
  • reverse transcription polymerase chain reaction
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Appears in Collections:Artigos, TCCs, Teses e Dissertações da Unesp

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