title: Functional modulation of enterocytes by gram-positive and gram-negative microorganisms.
abstract: Clinical studies have suggested that so-called probiotic bacteria may be effective as therapy in inflammatory bowel disease. However, the molecular mechanisms of their interaction with the intestinal surface remain undefined. The influence of whole probiotic bacteria [Escherichia coli Nissle 1917 (EcN); probiotic mixture VSL#3 (PM)], bacterial cell lysates, and conditioned media on transepithelial resistance (TER), IL-8 secretion, mucin gene expression, and tight junction proteins were determined in T84 and HT-29 intestinal epithelial cells (IEC). In addition, effects on pathogen (Salmonella dublin)-induced alterations were analyzed. EcN as well as debris and cell extracts induced IL-8 secretion from IEC, whereas no such effect was observed following incubation with the PM. The PM and soluble protein(s) released from the PM increased TER, prevented pathogen-induced decrease in TER, and were shown to stabilize tight junctions. The PM induced expression of mucins in IEC, and these organisms as well as EcN diminished S. dublin-induced cell death. Inhibition of MAPKs with PD-98059 or SB-203580 significantly decreased alterations in IL-8 synthesis and mucin expression and affected the regulation of TER. Probiotics and protein(s) released by these organisms may functionally modulate the intestinal epithelium of the host by different mechanisms, including the competition of whole organisms for contact with the epithelial surface as well as stabilization of the cytoskeleton and barrier function and the induction of mucin expression. Gram-negative and gram-positive organisms differ in the mechanisms activated, and a combination of organisms might be more effective than the application of a single strain.
authors.affiliation: Center for Study of Inflammatory Bowel Disease, Gastrointestinal Unit, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA. dpodolsky@partners.org
authors.forename: Jan-Michel
authors.initials: JM
authors.lastname: Otte
authors.forename: Daniel K
authors.initials: DK
authors.lastname: Podolsky
mesh_terms: D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D004396:Coloring Agents; D003470:Culture Media; D017097:Electric Impedance; D020895:Enterocytes / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006090:Gram-Negative Bacteria / Q000502:physiology*; D006094:Gram-Positive Bacteria / Q000502:physiology*; D006801:Humans; D007202:Indicators and Reagents; D016209:Interleukin-8 / Q000378:metabolism; D019936:Probiotics; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology; D019108:Tight Junctions / Q000502:physiology; D014343:Trypan Blue
chemical_list: D004396:Coloring Agents; D003470:Culture Media; D007202:Indicators and Reagents; D016209:Interleukin-8; D011506:Proteins; D012313:RNA; D014343:Trypan Blue