Co-incubation of PMN and CaCo-2 cells modulates inflammatory potential

Cell Mol Biol (Noisy-le-grand). 2017 May 20;63(5):119-126. doi: 10.14715/cmb/2017.63.5.22.


Polymorphonuclear granulocytes (PMN) are activated in inflammatory reactions. Intestinal epithelial cells are relevant for maintaining the intestinal barrier. We examined interactions of PMN and intestinal epithelial cell-like CaCo-2 cells to elucidate their regulation of inflammatory signalling and the impact of cyclooxygenase (COX), nitric oxide (NO) and platelet-activating factor (PAF). Human PMN and CaCo-2 cells, separately and in co-incubation, were stimulated with the calcium ionophore A23187 or with N-Formyl-methionyl-leucyl-phenylalanin (fMLP) that activates PMN only. Human neutrophil elastase (HNE) and respiratory Burst were measured. To evaluate the modulation of inflammatory crosstalk we applied inhibitors of COX (acetyl salicylic acid; ASA), NO-synthase (N-monomethyl-L-arginin; L-NMMA), and the PAF-receptor (WEB2086). Unstimulated, co-incubation of CaCo-2 cells and PMN led to significantly reduced Burst and elevated HNE as compared to PMN. After stimulation with A23187, co-incubation resulted in an inhibition of Burst and HNE. Using fMLP co-incubation failed to modulate Burst but increased HNE. Without stimulation, all three inhibitors abolished the effect of co-incubation on Burst but did not change HNE. ASA partly prevented modulation of Burst L-NMMA and WEB2086 did not change Burst but abolished mitigation of HNE. Without stimulation, co-incubation reduced Burst and elevated HNE. Activation of PMN and CaCo-2 cells by fMLP as compared to A23187 resulted in a completely different pattern of Burst and HNE, possibly due to single vs. dual cell activation. Anti-inflammatory effect of co-incubation might in part be due to due to COX-signalling governing Burst whereas NO- and PAF-dependent signalling seemed to control HNE release.

Keywords: CaCo-2 cells; Inflammation; Nitric oxide; Platelet-activating factor.; Polymorphonuclear granulocytes.

MeSH terms

  • Aspirin / pharmacology
  • Azepines / pharmacology
  • Caco-2 Cells
  • Calcimycin / pharmacology
  • Humans
  • Inflammation / pathology*
  • N-Formylmethionine Leucyl-Phenylalanine / pharmacology
  • Neutrophils / drug effects
  • Neutrophils / metabolism*
  • Neutrophils / pathology*
  • Pancreatic Elastase / metabolism
  • Respiratory Burst / drug effects
  • Triazoles / pharmacology
  • omega-N-Methylarginine / pharmacology


  • Azepines
  • Triazoles
  • WEB 2086
  • omega-N-Methylarginine
  • Calcimycin
  • N-Formylmethionine Leucyl-Phenylalanine
  • Pancreatic Elastase
  • Aspirin