title: Non-pathogenic bacteria elicit a differential cytokine response by intestinal epithelial cell/leucocyte co-cultures.
abstract: OBJECTIVE
  Intestinal epithelial cells (IEC) are thought to participate in the mucosal defence against bacteria and in the regulation of mucosal tissue homeostasis. Reactivity of IEC to bacterial signals may depend on interactions with immunocompetent cells. To address the question of whether non-pathogenic bacteria modify the immune response of the intestinal epithelium, we co-cultivated enterocyte-like CaCO-2 cells with human blood leucocytes in separate compartments of transwell cultures.
  
  
  METHODS
  CaCO-2/PBMC co-cultures were stimulated with non-pathogenic bacteria and enteropathogenic Escherichia coli. Expression of tumour necrosis factor alpha (TNF-alpha), interleukin (IL)-1beta, IL-8, monocyte chemoattracting protein 1 (MCP-1), and IL-10 was studied by enzyme linked immunosorbent assays (cytokine secretion) and by semiquantitative reverse transcription-polymerase chain reaction.
  
  
  RESULTS
  Challenge of CaCO-2 cells with non-pathogenic E coli and Lactobacillus sakei induced expression of IL-8, MCP-1, IL-1beta, and TNF-alpha mRNA in the presence of underlying leucocytes. Leucocyte sensitised CaCO-2 cells produced TNF-alpha and IL-1beta whereas IL-10 was exclusively secreted by human peripheral blood mononuclear cells. CaCO-2 cells alone remained hyporesponsive to the bacterial challenge. Lactobacillus johnsonii, an intestinal isolate, showed reduced potential to induce proinflammatory cytokines but increased transforming growth factor beta mRNA in leucocyte sensitised CaCO-2 cells. TNF-alpha was identified as one of the early mediators involved in cellular cross talk. In the presence of leucocytes, discriminative activation of CaCO-2 cells was observed between enteropathogenic E coli and non-pathogenic bacteria.
  
  
  CONCLUSIONS
  The differential recognition of non-pathogenic bacteria by CaCO-2 cells required the presence of underlying leucocytes. These results strengthen the hypothesis that bacterial signalling at the mucosal surface is dependent on a network of cellular interactions.
authors.affiliation: Institute of Biological Chemistry and Nutrition Science, University Hohenheim, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Haller
authors.forename: C
authors.initials: C
authors.lastname: Bode
authors.forename: W P
authors.initials: WP
authors.lastname: Hammes
authors.forename: A M
authors.initials: AM
authors.lastname: Pfeifer
authors.forename: E J
authors.initials: EJ
authors.lastname: Schiffrin
authors.forename: S
authors.initials: S
authors.lastname: Blum
mesh_terms: D001419:Bacteria / Q000276:immunology*; D001422:Bacterial Adhesion; D018938:Caco-2 Cells; D002450:Cell Communication / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004847:Epithelial Cells / Q000276:immunology; D004926:Escherichia coli / Q000472:pathogenicity; D015870:Gene Expression; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis; D007413:Intestinal Mucosa / Q000276:immunology* / Q000382:microbiology*; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D016207:Cytokines; D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha