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33 TLR3 and interferon function in human embryonic stem cell-derived endothelial cells (hESC-EC): relevance to viral immunity
  1. D M Reed1,
  2. G Foldes1,
  3. R V Badiger1,
  4. N N Ali1,
  5. C P Wheeler-Jones2,
  6. M J Paul-Clark1,
  7. S E Harding1,
  8. J A Mitchell1
  1. 1National Heart and Lung Institute, Imperial College London, London, UK
  2. 2The Royal Veterinary College, University of London, London, UK


Human embryonic stem cell-derived endothelial cells (hESC-EC) are being investigated as research tools and cell therapy for vascular inflammation. We have shown that hESC-EC do not respond to certain bacteria pathogen associated molecular patterns (PAMP). Endothelial cells also respond to viral PAMPs via toll-like receptor-3 (TLR3) which couples with IRF transcription factors. IRF can also be activated by interferons (IFN) which are also released during immune reactions. We have compared PolyIC (10 μg/ml; TLR3 agonist), and IFN (30 ng/ml) responses in hESC-EC and human umbilical vein endothelial cells (HUVEC). CXCL8 and IP10 were measured by ELISA as biomarkers for NF-κB and IRF pathways respectively. All data are % control (100) ± SEM n=5–6. Both hESC-EC (IFNα; 289.8±71.6, IFNβ; 656.8±182.3, IFNγ; 878.6±275.9, IFNλ 55.1±18.7) and HUVEC (IFNα; 1419.3±445.8, IFNβ; 3144.8±946.6, IFNγ; 2982.7±583.9, IFNλ; 132.8±30.1) released IP10 in response to each IFN (p<0.05; one-sample t test) except IFNλ (p>0.05; one-sample t test). HUVEC (3593.4±1269.4) but not hESC-EC (73.3±12.0) also responded to PolyIC with release of IP10. CXCL8 was not elevated by any treatment in either cell type. hESC-EC and HUVEC have a functional IRF transcription factor pathway that is activated in response IFNα, β and γ but not IFNλ. hESC-EC do not have a detectable TLR3 function. These findings may have implications for the use of hESC-EC in research and therapy and a more thorough investigation of these pathways is needed.

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